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Full text of "The Pfizer handbook of microbial metabolites"

3a^^^^^^^E3^^^^^E3[ 



] Marine Biological Laboratory Library I 

Q Woods Hole, Mass. 



Presented by 

Chas, Pfizer and Co., Inc. 
Medical Research Lab, 
Groton, Conn* 



I 
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THE 

PFIZER HANDBOOK 

OF 

MICROBIAL METABOLITES 



/■'K 6 



THE 

PFIZER HANDBOOK 

OF 

MICROBIAL 
METABOLITES 



By 

MAX W. MILLER, ph.d. 

Pfizer Medical Research Laboratories, 
Chas. Pfizer & Co., Inc. 




The Blakiston Division 

McGRAW-HILL BOOK COMPANY, INC. 

New York Toronto London 



THE PFIZER HANDBOOK OF MICROBIAL METABOLITES 

Copyright © 1961 by Chas. Pfizer & Co., Inc. Printed in 
the United States of America. All rights reserved. This 
book, or parts thereof, may not be reproduced in any 
form without permission of the copyright owner. 



Library of Congress Catalog Card Number: 61-17138 
49755 



Foreword 



The impressive advances achieved in fermentation techniques 
have created new and often highly efficient methods for the 
synthesis of organic compounds. It seems clear that in addition 
to antibiotics and steroids, an ever-increasing number of struc- 
turally less complicated chemicals will be synthesized most eco- 
nomically by fermentation of abundant starting materials of 
natural or synthetic origin. 

The purpose of this handbook is to list the source and physi- 
cal, chemical and physiological properties of metabolic products 
isolated from bacteria, molds, fungi and lichens. In addition to 
this collection of facts and references, it contains chapters out- 
Uning the biogenesis of various structural types elaborated 
mainly by microorganisms. Although some of our present-day 
views on biogenetic pathways may have to be revised in the fu- 
ture, these chapters should prove to be exceedingly helpful not 
only to chemists working on the structures of new substances 
but also to biochemists investigating the mode of action of 
physiologically active compounds. 

There certainly was an urgent need for such a compilation be- 
cause the original reports are scattered through a wide variety 
of scientific journals rarely assembled in one place but distrib- 
uted in chemical, pharmaceutical and medical libraries. It 
seems highly appropriate that an attempt to cover the literature 
in this rapidly expanding field should come from the Research 
Division of Chas. Pfizer & Co., Inc. The group deserves a great 
deal of credit for pioneering work in industrial fermentation 
as well as in isolation and structure elucidation of many anti- 
biotics. 

G. BucHi 

Cambridge, Massachusetts 



Acknowledgment 



A COMPILATION of this soit was suggested by Dr. Ernest M. 
Weber in 1956, and the first draft was issued as an intra- 
company report the following year. Later, publication was sug- 
gested by Dr. Gilbert M. Shull and urged by a number of uni- 
versity people interested in microbial metabolites. 

Most importantly, publication would not have been possible 
without the consent and support of Dr. Karl J. Brunings and 
Dr. I. A. Solomons. Other staff members of the Pfizer Medical 
Research Laboratories have also been very cooperative. Dr. 
Frank A. Hochstein has been most helpful throughout the prepa- 
ration for publication, and I wish to thank him especially as 
well as Dr. Walter D. Celmer for reading the manuscript at an 
early stage and for their comments on the chapter on macrolide 
antibiotics. 

In addition. Dr. Francis X. Murphy read the entire galley 
proof and made many constructive suggestions. 

Several other authorities have been kind enough to review 
their specialties. Professor Hans Brockmann of Gottingen con- 
tributed information on the actinomycins; Professor Konrad 
Bloch of Harvard read the sections dealing with lipides; Dr. 
T. G. Halsall of Oxford reviewed fungal steroids; Dr. Herchel 
Smith of Manchester, sections concerned with the biosynthesis 
of various mold metabolites; Professor F. G. Holliman of Cape- 
town, the section on phenazines; Dr. J. D. Bu'Lock of Man- 
chester, the section on acetylenic substances; and Dr. Edward 
Borowsky of the Institut Medycyny Moskiej, Gdansk, the sec- 



Acknowledgment viii 

tion on polyene macrolides. Professor George Biichi of Massa- 
chusetts Institute of Technology read nearly all of the galley 
proof and contributed a generous foreword. 

We cannot begin to acknowledge all of the assistance re- 
ceived, particularly from the Pfizer library staff and other 
libraries, from our colleagues on the chemical staff, and from 
the secretarial staff. Most of the manuscript typing was done by 
Miss Kathryn Beck, Mrs. Loretta Michaud, Mrs. Terry Lunt, 
Mrs. Hedy Korst, Mrs. Judith Neff, and Miss Patricia Goepfert. 
The references were corrected and much indexing was done by 
Miss Claudette Parent, Miss Grace Olimski, and Miss Patricia 
French. AU of the copy-editing was done by Mrs. Margaret 
Thompson. Patricia Curtis of Editorial Projects, Inc. was very 
helpful in coordinating and expediting pubUcation operations. 

Max W. Miller 
Groton, Connecticut 




Contents 



Introduction 

1. Simple Hydrocarbons, Ketones, Aldehydes, 

Esters, etc. 9 

2. Alcohols, Glycols and Compounds Related to Sugars 13 

3. Aliphatic Acids and Glycolipides 46 

4. Tetronic Acids and Other Lactones and Lactams 79 

5. Carotenes and Carotenoids 90 

6. Polyenes and Polyynes, Excluding 

Polyene Macrolides 107 

7. Macrocyclic Lactones (Macrolides) 118 

a. POLYENE MACROLIDES 123 

b. OTHER MACROLIDES 130 

8. Alicyclic Compounds Other Than Terpenoids and 
Steroids 142 

9. Terpenoids and Steroids 154 

10. Tropolone Acids 181 

11. Phenolic Substances 185 

a. PHENOLS AND PHENOL ETHERS (GENERAL) 185 

b. DEPSIDES AND DEPSIDONES 212 

12. Quinones and Related Compounds 231 

a. BENZOQUINONES 239 

b. NAPHTHOQUINONES 248 
C. ANTHRAQUINONES 254 

13. Tetracycline, Analogues and Related Substances 273 

14. Aromatic Compounds Not Classified Elsewhere 284 

15. Amines 290 

16. Amino Acids and Related Compounds 299 

17. Polypeptides and Related Compounds 332 

18. Heterocycles 398 

a. FURANS AND RELATED SUBSTANCES 398 

b. DIBENZOFURANS AND RELATED SUBSTANCES 400 
C. PYRANS AND RELATED SUBSTANCES 404 
d. XANTHONES 416 



Contents x 

e. COMPOUNDS RELATED TO THIOPHENE, IMIDAZOLE, 
THIAZOLE AND ISOXAZOLE 418 

f. PYRROLES, PORPHYRINS AND RELATED 
COMPOUNDS 434 

g. INDOLES 458 

h. ERGOT ALKALOIDS 465 

i. PYRIDINES 479 

j. QUINOLINES 492 

k. PYRAZINES, DIKETOPIPERAZINES 496 

I. PHENAZINES AND PHENOXAZONES 501 

m. PYRIMIDINES 508 

n. PURINES 524 

O. PTERIDINES AND FLAVINES 548 

19. Unclassified Metabolites 572 

Bibliography, Reviews and General References 615 
Appendixes 

A. Chemical Compositions of the Tissues and Large 

Molecules of Bacteria and Fungi 623 

B. Bacterial and Fungal Carotenes 638 
c. The Chemical Constituents of Mycobacteria 645 

Addendum 661 

Subject Index 715 

Empirical Formula Index 748 

Microorganism Index 758 



THE 

PFIZER HANDBOOK 

OF 

MICROBIAL METABOLITES 



Introduction 



The culture of bacteria and molds, the collection of higher 
fungi and lichens and the isolation and characterization of their 
metabolites is a sophisticated sort of research involving several 
distinct sciences. As a result the reports of such work are scat- 
tered through a variety of chemical, biochemical, microbiologi- 
cal, botanical, medical and pharmaceutical journals as well as 
general scientific journals and those devoted to antibiotics and 
fermentation technology. The published reviews of the struc- 
tures of microbial metabolites have been Umited in scope. 
It is difficult for the novice to gain a total impression of the 
progress that has been made, and difficult even for the specialist 
in this area to see the forest entire as well as the trees about him. 

Having monitored the literature for several years incidental 
to our own work, we felt that it would be useful to publish a 
more general list of chemicals produced by microorganisms. 
More specifically, what has been attempted is a compilation of 
data on the structural and simpler physical properties of all of 
the primary microorganism metabolites which have been re- 
ported to be produced by the organisms growing either in the 
wild state or in culture on artificial sugar-based media. Al- 
though many structures are incomplete, generally the com- 
pounds in this list have been purified, and at least some physical 
properties observed. In view of the difficulties mentioned above 
we do not presume to have achieved absolutely complete cover- 
age, and we should be pleased to receive structures or references 
to appropriate compounds which have been overlooked. Cor- 
rections of errors would be appreciated also. The literature 
available to us has been watched until the beginning of printing 
operations in December 1960. 

Organization is by general similarity of chemical structures, 
but not in the strictest sense. For example, all carotenes and 
carotenoids were grouped together rather than grouping a caro- 



Pfizer Handbook of Microbial Metabolites 4 

tene alcohol with, e.g., a steroid alcohol. Many substances are 
ambiguous and could have been classified in any of several dif- 
ferent chapters. A substance which contains a sugar, a benzene 
ring, a terpenoid fragment and a heterocycle will most likely be 
found under the appropriate heterocycle classification. Some 
arbitrary decisions have been necessary, but indexing by name, 
by empirical formula and by producing microorganism should 
serve most purposes. Again quite generally, progression is from 
the simple to the complex; sugarlike compounds being con- 
sidered simple because they resemble the substrate, glucose. 

In order to make the list more coherent a background has 
been sketched in, emphasizing occurrence and biosynthetic 
origin. A considerable literature on the biosynthetic origin of 
microbial metabolites has accumulated. Familiarity wdth it is 
valuable in interpreting experimental results in structure deter- 
minations. Several old structures have been revised in the light 
of this new knowledge. 

Many of the biosynthetic and other metabolic schemes worked 
out in microorganisms are quite general in occurrence and have 
been found to be operative in mammalian metabolism. Be- 
cause bacteria and fungi grow rapidly and are easy and inexpen- 
sive to handle, they are among the most useful tools in the ex- 
ploration of metabolic routes. Many of the chemicals in this 
list were isolated incident to such studies. 

Some chemicals of metabolic significance and of a suitable 
degree of complexity can be produced economically in quantity 
by fermentation methods and have found industrial uses. An 
example is citric acid, which now finds an annual market of 
thousands of tons. 

The discovery of the effectiveness of the mold product, peni- 
cilHn, in treating many bacterial infections in man gave tremen- 
dous impetus to the isolation and screening of microorganisms 
and their metabolites for antibiotics. The isolation and study 
of microbial metabolites, formerly a scholarly pursuit in a few 
academic laboratories, suddenly was supported by the resources 
of a great industry. Experience showed that a genus of filamen- 
tous soil organism, the actinomycete ( streptomycete ) , was a 



Introduction 



particularly prolific source of organisms adaptable to antibiotics 
production when grown in suitable media. 

Research with the actinomycetes resulted in the discovery of 
agents effective against a broad spectrum of pathogens. The 
first of these were chloramphenicol, chlortetracycline and oxy- 
tetracycline. Since the discovery of oxy tetracycline, no anti- 
biotics of broader antibacterial range have been developed. 

Prior to the discovery of antibiotics, much work had been 
done on the structures of lichen substances, and, as mentioned 
above, a few academic laboratories were interested in mold 
metabolites. Notable among these was Professor Harold 
Raistrick's group at the London School of Hygiene and Tropical 
Medicine. Raistrick, now retired, and his collaborators have 
published over 100 papers on this topic. 

The academic investigators were impelled by no practical 
motive except perhaps a hope that comparison of the chemical 
metabolites of various ill-defined groups of fungi would assist 
in their classification. Some generalizations did become ap- 
parent, but on the whole this hope was disappointed. It was 
found that the same chemical might even be produced by both 
bacteria and fungi. Some of the old classification schemes 
based on pigmentation were found to be obsolete. 

The structures of the large molecules produced by micro- 
organisms have proved to be more specific and of real value to 
taxonomy. Since the advent of paper chromatography, the 
identification of amino acids, sugars and other fragments from 
cell tissue hydrolysates has been facilitated. From the ensuing 
proliferation of literature on this subject it is manifest that the 
compositions of various cell tissues (capsule, wall, protoplast 
membrane, internal proteins), as well as exotoxins and other 
high molecular weight exudates, are much more specific. Even 
strains of species can sometimes be distinguished by the pres- 
ence or absence of one of these fragments, and these molecules 
are important in immunology. Work of this sort has become 
more important since the discovery of evidence that certain 
antibiotics, e.g., penicillin, interrupt growth and cell division in 
the bacteria against which they are effective by interfering with 



Pfizer Handbook of Microbial Metabolites 6 

normal cell wall synthesis. Although we were unable to pursue 
this fascinating topic, an appendix of literature titles on the 
structure of higher molecular weight products of microorgan- 
isms and their cell wall structures has been attached. 

In comparing the structures of the hundreds of microorgan- 
ism metabolites which have been characterized thoroughly it is 
well to remember that the statistical emphasis may be mislead- 
ing. It is likely that insoluble compounds, lipophilic materials 
easily extractible from aqueous cultures, organic acids which 
can be precipitated as insoluble salts and pigments that are 
easily observed have probably received a disproportionate degree 
of attention. The same, of course, could be said for antibiotics, 
which are conspicuous for their biological activity. The most 
difficultly discoverable metabolites are the relatively inconspicu- 
ous, low molecular weight, hydrophilic, perhaps phosphorylated 
compounds. Eventually many of the precursors of more elabo- 
rate metabolites will be found in this category. 

Also, the metabolites of certain microorganisms have re- 
ceived disproportionate study. Examples are Mycobacterium 
tuberculosis, the tuberculosis pathogen, and Claviceps purpurea, 
the ergot fungus. By permission of Dr. Esmond R. Long and 
the Williams and Wilkins Publishing Company a review of the 
known metabolites of the former organism has been reproduced 
as an appendix, although many of the compounds included in 
this review are also to be found in the body of the text and others 
in the text which were not in the review. Also an appendix 
dealing with the confusing subject of microbial carotenoids has 
been attached by permission of the Chemical Publishing Com- 
pany and of Professor T. W. Goodwin of the University of Liver- 
pool. 

Referencing is not exhaustive. It was kept on the lean side 
intentionally, and we feel that it is more useful that way. On 
some topics the literature is vast. It would have been virtually 
impossible to offer complete referencing of, for example, acetic 
acid, or even of some of the more complex substances such as 
the gibberellins or ^-carotene. Much attention has been given 
to choice of useful references, although no doubt there have 



Introduction 



been lapses, and differences of opinion will probably arise. For 
some of the substances carrying a large literature a review 
article often is cited. In general an attempt has been made to 
cite the isolation, final structure determination and synthesis 
papers insofar as they exist. In the references cited care has 
been taken to include the complete list of authors as given on 
the paper. A bibliography of books, general references and re- 
views is included at the end. 

Occasional comments may be found at the bottom of an entry, 
reflecting the manner in which this material evolved from a 
card file with a few notes. These comments were allowed to 
stand without expansion for what they are worth. For the most 
part the work is uncritical, structures and properties having 
been transcribed just as given in the literature. Structures and 
empirical formulas designated as tentative or approximate by 
the authors have been so designated here. 

The indexes were not available prior to printing, and it is 
hoped that they will point out hitherto unrecognized relation- 
ships. 



Simple Hydrocarbons, Ketones, 
Aldehydes, Esters, etc. 



The simple compounds listed here cannot be treated as 
a class. The biogenetic origins of many of them should 
become apparent from the introductions to later chapters. 
Besides the hydrocarbons shown it might be mentioned 
that lactarius species sporophores contain cis-polyiso- 
prene, a rubber-like substance. 

W. D. Stewart, W. L. Wachtel, J. J. Shipman and J. A. 
Yanko, Science 122 1271 (1955). 

1 Thiourea, CH4N2S, white crystals, m.p. 180-182°. 

S 
H2N— C— NH2 

Verticillium albo-atrum, Botrytis cinerea 

K. Ovcharov, Compt. rend. acad. sci., U.S.S.R. 16 461 

(1937). 

2 Guanidine, CH5N3, alkaline crystals, generally isolated as salts, 

e.g. acetate, m.p. 229°. 

NH 

H2N— C— NH2 

Boletus edulis, Hydnum aspratum Berk. 
E. Winterstein, C. Reuter and R. Korolev, /. Chem. Soc. 104 
433 (1913). 

Seijiro Inagaki, J. Pharm. Soc. Japan 54 824 (1934). 

3 Ethylene, C2H4, colorless gas, b.p. —103°. 

CH2=CH2 



Pfizer Handbook of Microbial Metabolites lO 

Penicillium digitatum, Blastomyces dermatitidis , B. bra- 
siliensis, Histoplasma capsulatum 

Walter J. Nickerson, Arch. Biochem. 17 225 (1948). 

Erston V. Miller, J. R. Winston and D. F. Fisher, J. Agr. 
Research 60 269 (1940). 

Ray E. Young, Harlan K. Pratt and J. B. Biole, Plant 
Physiol. 26 304 (1951). 

4 Dimethylsulfone, CoHgOoS, colorless prisms, m.p. 107-109°. 

CH3SO2CH3 

Cladonia deformis Hoffm. 

Torger Bruun and Nils Andreas Sorensen, Acta Chem. 
Scand. 8 703 (1954). 

5 Cellocidin (Aquamycin), C4H4O2N0, white crystals, m.p. 216- 

218° (dec). 

O O 

li II 

HoN— C— C=C— C— NH2 

Streptomyces chibaensis, S. reticuli var. aquamyceticus 

The yield was 16.5 g. from 420 liters of culture fluid. 

Saburo Suzuki, Goto Nakamura, Kazuhiko Okuma and Yoke 
Tomiyama, J. Antibiotics (Japan) llA 81 (1958). 

Hyozo Taniyama, Shoji Takemura, Kimiko Kageyama and 
Masanao Funaki, /. Pharm. Soc. Japan 79 1510 (1959). 

6 Ethyl Acetate, C^HsOo, colorless liquid, b.p. 77°, 11^^° 1.3719. 

CH3COOC2H6 

PenicilliuTn digitatum 

J. H. BirkinsHaw and H. Raistrick, Trans. Roy. Soc. (Lon- 
don) B220 331 (1931). 

7 2-Methyl-2-butene, C5H10, colorless liquid, b.p. 38.4°. 

CH3 

\ 

C=CH— CH3 

/ 
CH3 

Puccinia gram,inis Pers. var. tritici Erikas. and Henn. 
(uredospores) 

F. R. Forsyth, Can. }. Botany .S3 363 (1955). 



1 1 Simple Hydrocarbons, Ketones, Aldehydes, Esters, etc. 

8 l-Ethoxy-l,2-ethylenedicarboxamide, CcH,oO;^N2, 

O O 

II II 

HoN— C— C=CH— C— NH2 

OC0H5 

Streptomyces sp. 

Yasuharu Sekizawa, /. Biochem. Japan 45 73 (1958). 

9 Isobutyl Acetate, CfiHjoOs, colorless liquid, b.p. 61°, Uo^^ 1.3936. 

O CH3 

II / 

CH,— C— O— CH2— CH 

\ 
CH3 

Endoconidiophora coerulescens 

J. H. Birkinshaw and E. N. Morgan, Biochem. J. 47 55 
(1950). 

10 2-Methyl-2-heptene-6-one, CsHi^O, colorless liquid, b.p. 172-174°, 
58° (10 mm.), n,r" 1.4445. 

O CH3 

II / 

CH3— C— CH2— CH2— CH=C 

\ 
CH3 

Endoconidiophora coerulescens Miinch, E. virescens 
Davidson (artificial medium) 

Isobutyl acetate and a mixture of methylheptenols were 
isolated from the same culture. 

J. H. Birkinshaw and E. N. Morgan, Biochem. J. 47 55 
(1950). 
n Octacosane, CogHsg, colorless crystals, m.p. 61°. 

CH3(CH,),bCH3 

Amanita phalloides 

Heinrich Wieland and Gustav Coutelle, Ann. 548 270 
(1941). 

12 Actinomycin J2 (Waksman's Actinomycin B, Dodecyl Ester of 
5-Oxostearic Acid), C^Jir^gO^, colorless crystals, m.p. 
81.5°. 

CH3(CH2)l2CO(CH2)3COOCl2H28 



Pfizer Handbook of Microbial Metabolites 12 



Actinomyces (Streptomyces) fiavus 

Yoshimasa Hirata and Koji Nakanishi, Bull. Che-m. Soc. 
Japan 22 121 (1949). 

13 cts-Palmitenone, CgiHeoO, colorless microcrystals, m.p. 40°. 

O 
CH3(CH2)i4— C— (CH2)7— CH=CH— (CHslaCHs 

Corynebacterium diphtheriae 

J. Pudles and E. Lederer, Biochim. et Biophys. Acta 11 602 
(1953). 

Idem., Bull. soc. chim. biol. 36 759 (1954). 

14 Palmitone, CgiHgoO, colorless leaflets, m.p. 82°. 

O 

II 
CH3(CH2)i4— C— (CHsluCHs 

Corynebacterium diphtheriae 

J. Pudles and E. Lederer, Bull. soc. chim. biol. 36 759 
(1954). 



Alcohols, Glycols and Compounds 
Related to Sugars 



Two of the most important routes of sugar metabolism are 
the Embden-Meyerhof pathway of anaerobic glycolysis and the 
oxidative pentose phosphate cycles. Both occur widely in nature, 
and microorganisms were useful in the discovery of each. Many 
of the metabolites of this chapter can be pictured as arising 
from one of these schemes, which are also the main known 
routes of glucose metabolism in mammals. It should be under- 
stood that other paths and fragments of paths of glucose metab- 
olism have been found in various microorganisms. 

Yeast was instrumental in the elucidation of the Embden- 
Meyerhof route^ and the yeast alcohol fermentation is repre- 
sented as follows, each step catalyzed by a specific enzyme: 

Embden-Meyerhof Route of Anaerobic Glycolysis in Yeast 

Enzymes 

1. Hexokinase 

2. Phosphohexoisomerase 

3. Phosphohexokinase 

4. Aldolase 

5. Triosephosphate isomerase 

6. Triosephosphate dehydrogenase (Inhibited by iodoacetate) 

7. ATP-Phosphoglyceric transphosphorylase 

8. Phosphoglyceromutase 

9. Enolase (Inhibited by fluoride) 

10. ATP-Phosphopyruvic transphosphorylase 

11. Carboxylase 

12. Alcohol dehydrogenase 

^ A. J. Kluyver and C. B. Van Niel, "The Microbe's Contribution to 
Biology," Harvard University Press, Cambridge, Massachusetts, 1956. 



Pfizer Handbook of Microbial Metabolites 



14 



Glucose 



C 



ATP 
ADP 



Glucose-6-phosphate 

jr 

Froctose-6-phosphate 



C 



ATP 
ADP 



Fructose-l,6-diphosphate 

l\ 

D-Glyceraldehyde-3-phosphafe ;:i Dihydroxyacetone Phosphate 

-11 /*DPN© 
H3PO4 11 V^DPNH + H© 

D-1, 3-Diphosphoglyceric Acid 

D-3-Phosphoglyceric Acid 

11 
D-2-Phosphoglyceric Acid 



H2O 

Phosphoenoipyruvic Acid 

"- S.ATP 
Pyruvic Acid 



Acetaldehyde + CO2 

11 /*DPNH + H© 
1^ ^DPN© 



Ethanol 



Many molds, actinomycetes and bacteria use this system to 
some degree. Variations occur, and intermediates may feed in 
from other sources, for example, triose phosphate from the pen- 



1 5 Alcohols, Glycols and Compounds Related to Sugars 

tose phosphate cycle. Some bacteria are able to produce alcohol 
by other means. 

The pyruvate from anaerobic glycolysis can meet a variety of 
fates. In some cases it is transformed into acetoin and its oxi- 
dation and reduction products, diacetyl and 2,3-butanediol 
(thiamine pyrophosphate coenzyme). ct-Acetolactic acid has 
been shown to be an intermediate in certain instances :- 



•2CO2 



2CH3COCOOH - 




* CH3CHO 1 TPP 


* CH3CHO 




-CO2 


/ 






COOH 

II 1 
CH3C— C— CH3 — 

1 


-CO2 


° / 
II / 
-♦ CH3— C— CH— CH3 

1 






1 
OH 




1 
OH 






o-Acetolactic Acid 




Acetoin 

1 








CHs— CH- 


-CH- 


H2 
CH3< 




[01 

»CH3- 




II 
-C- 




II 
-C— CH3 



OH OH 
2,3-Butanediol 



Diacetyl 



Acetoin has been found in yeast, in other fungi and in bac- 
teria. Large yields of mixtures of these condensation products 
can be obtained from some bacteria. 

Pyruvate is reduced to D-lactic acid in the homofermentative 
bacteria and lower phycomycetes (and to L-lactic acid in mam- 
malian muscle). 

Another reaction of pyruvate is its conversion to acetylcoen- 
zyme A with the participation of lipoic acid; the probable me- 
chanism being :^ 



2 Elliot Juni, /. Biol. Chem. 195 715 (1952); Yutaka Kobayashi 
and George Kalnitsky, ibid. 211 473 (1954). 

^ I. A. Gunsalus, Lois S. Barton and H. Gruber, /. Am. Chem. Soc. 
78 1763 (1956). 



Pfizer Handbook of Microbial Metabolites 



i6 



CH3COCOOH 




TPP -^ 
Mg©> |S> CO2 




CH3CHO— \Thiamine 

J Pyrophosphate (TPP) 




TPP ^ — y ^\ 




1 


-S 

1 


CH2— CH2- 


-CH(CH2)4COOH 


■ r 

Lipoic Acid 
SH S— COCH3 \ 

1 1 > 


^->DPNH + H© 


CH2CH2CH(CH2)4COOH 


V > DPN© 


5-AcetyIdihydrolipoic Acid 


\ 



Coenzyme A 




SH 

I 
CH2— CH2— CH(CH2)4COOH 

Dihydroiipoic Acid 

CH3CO — Coenzyme A 



The nature of the actual catalysis of pyruvate decarboxyla- 
tion and of aldol condensations by thiamine pyrophosphate co- 
enzyme has been elucidated.^ It is shown below : 



NH2 



// 



Thiamine 

Pyrophosphate 
Chloride 



CH3 




O O 

CH2— N' I t T 

<S^=^CH2— CH2— O— P— O— P— OH 



CHa 



^.e 



e, 



OH 

CH3— c— cooQ 



OH 







@/^S -H® ®/^~S CH3— C— COO0 ©/^S -CO2 

R— N I »R— N; I >R— N/ ' 



OH 



CHi 



R— N 



/~s 




O OH 



CH3— C— C— 



— C— 

* Ronald Breslow, Chem. and Ind., 893 (1957). 



I? 



Alcohols, Glycols and Compounds Related to Sugars 



Thus, the production of acetaldehyde (and subsequently al- 
cohol) by yeast, the production of acetoin by certain bacteria, 
etc. 

Although the lipoic acid mechanism was first demonstrated 
in Streptococcus faecalis, all bacteria do not require the cofac- 
tor for this transformation. 

The role of acetylcoenzyme A in cellular synthesis of fatty 
acids will be seen later. Butanol is probably formed by reduc- 
tion of acetoacetylcoenzyme A. It is interesting to note that 
some microorganisms can synthesize a variety of carbohydrates 
by using acetate as the sole carbon source, in effect reversing 
the process (e.g.^). Pyruvate is also converted to succinate by 
fixation of COo. 

Various other fates of pyruvate are known. For example, 
there are bacteria which dismutate 2 moles of pyruvate to 
1 mole each of acetic and lactic acids. *^ Also Bacillus coli is 
known to convert pyruvate to a mixture of acetic and formic 
acids. ^ 

The pentose phosphate cycle mentioned earlier probably oc- 
curs in many microorganisms. It is outlined below: 

Ribulose-5-P < » Ribose-5-P 



TPNH 

+H© 



TPN© 




CO2 

6-Phosphogluconic 

Acid \ >r TPNH 

+ H© 



Xylulose-5-P 




Glucose 



ATP 



Dihydroxy- 
Qcetone-P 



Glyceralde 
hyde-3-P 



♦ Fructose-6-P 



Glyceralde- 
hyde-3-P 




Sedoheptu- 
ose-7-P 



Erythrose-4-P 



Xylulose-5-P 



Enzyme-catalyzed reactions of the pentose 
phosphate pathway* 

* This diagram together with the summarizing equations is re- 
printed with permission from Joseph S. Fruton and Sofia Simmonds, 
"General Biochemistry," John Wiley and Sons, Inc., New York, N. Y., 
1958, p. 531. 

5 V. I. Lyubimov, Doklady Akad. Nauk SSSR III No. 4 (1956). 

" Seymour Karkes, Alice del Campillo, I. C. Gunsalus and Severe 
Ochoa, J. Biol. Chem. 193 721 (1952). 

^ Kenneth V. Thimann, "The Life of Bacteria," Macmillan Co., 
New York, N. Y. 1955, pp. 441-465. 



Pfizer Handbook of Microbial Metabolites i8 

These reactions in summary are: 

6 Hexose phosphate + 60.- — > 6 Pentose phosphate + 6CO> + 6H2O 
4 Pentose phosphate — > 2 Hexose phosphate + 2 Tetrose phosphate 
2 Pentose phosphate + 2 Tetrose phosphate — > 

2 Hexose phosphate + 2 Triose phosphate 
2 Triose phosphate + H2O — > Hexose phosphate + phosphate 

Hexose phosphate + 6O2 — > 6CO.; + 5H2O + phosphate 

This is, then, a route for the complete degradation of glucose 
to carbon dioxide and water. The statistical significance and 
prevalence of this oxidative degradation system among micro- 
organisms remains to be determined. 

Ribose can be synthesized by way of the pentose phosphate 
cycle. In B. coli it appears that deoxyribose arises from direct 
reduction of ribose.® 

Gluconic acid occurs widely, especially in fungi, and can be 
formed by enzyme-catalyzed oxidation of the unphosphorylated 
glucose substrate.'* In some oxidative bacteria the following 
scheme occurs:^" 

Glucose — > Gluconic Acid — > 6-Phosphogluconic Acid — > 

2-Keto-3-deoxy-6-phosphogluconic Acid -^ Pyruvic Acid 

+ 
Glyceraidehyde-3-phosphate 

The glyceraldehyde phosphate is easily convertible to another 
mole of pyruvic acid. 

Both glucuronic acid" and fucose (6-deoxy-L-galactose)^'- 
seem to be formed from glucose without cleavage of the carbon 
skeleton. 

Glucosamine is probably most commonly formed by gluta- 
mine amination of fructose-6-phosphate,^'^ although glucosone 

^ Fillmore K. Bagatell, Elmer M. Wright and Henry Z. Sable, 
;. Biol. Chem. 234 1369 (1959). 

" Vincent W. Cochrane, "Physiology of Fungi," John Wiley and 
Sons, Inc., New York, N. Y. 1958, pp. 131-135. 

1" Nathan Entner and Michael DoudorofF, /. Biol. Chem. 196 853 
(1952); Joseph MacGee and Michael DoudorofF, ibid. 210 617 (1954). 

" Frank Eisenberg, Jr. and Samuel Gurin, J. Biol. Chem. 195 317 
(1952); Frank Eisenberg, Jr., ibid. 212 501 (1955). 

1- J. F. Wilkinson, Nature 180 995 (1957); Stanton Segal and Yale 
J. Topper, Biochim. et BiopJiys. Acta 25 419 (1957). 

1^ Luis F. Leloir and Carlos E. Cardini, Biochim.. et Biophys. Acta 
12 15 (1953). 



19 



Alcohols, Glycols and Compounds Related to Sugars 



(a logical precursor) has been shown to be formed by some 
aspergilli. 

Mannitol, which is accumulated in quantity by some micro- 
organisms and occurs widely, is known in some cases to be in a 
reversible equilibrium with fructose, and it probably serves as a 
reserve food." This reserve function may be true also of other 
reduced sugars. 

The inositols are not formed by direct hexose cyclization, but 
their detailed biosynthesis is not known. 

Many uncommon sugars have been found as moieties of 
streptomycete antibiotics. Some of these antibiotics which are 
predominantly sugar-like in composition are included at the 
end of this chapter. It might be useful to list the individual 
sugars here for comparison, including those which occur in 
streptomycete antibiotics classified in other chapters : 

Sugars from Streptomycete Antibiotics 

(showing points of attachment and 

stereochemistry where known) 




N-Methyl-i-glucosamine 
(streptomycins) 



HsC 




Streptose 
(streptomycin) 




OH 
Streptidine 
(streptomycin) 




CH2NH 



Dihydrostreptose 
(dihydrostreptomycin) 




HOCH 



Hydroxystreptose 
(hydroxystreptomycin) 




6-Giucosamine 
(kanamycin) 



^* Vincent W. Cochrane, "Physiology of Fungi," John Wiley and 
Sons, Inc., New York, 1958, p. 122. 



Pfizer Handbook of Microbial Metabolites 



20 



CH2OH 




Kanosamlne 
(kanamycin) 




2-Deoxystreptamine 
(kanamycin, paromomycin) 



CH2OH 




D-Glucosamine 
(paromomycin, 
trehalosamine) 



CH2NH 




HO 'NH2 

Paromose 
(paromomycin) 



(Neosamine C from 
neomycin is also a 
2,6-diaminohexose.) 




H2N OH 

Mycosamine 

(nystatin, 

amphotericin B, 

pimaricin) 




(CH3)2N 

Desosamine 

or 

Picrocin 

(picromycin, methy- 
mycin, neomethymy- 
cin, narbomycin, ole- 
andomycin, erythro- 
mycins A, B, and C.) 



CH3O 

Oleandrose 
(oleandomycin) 




CH3O 

Cladinose 
(erythromycins A, B) 



2 1 Alcohols, Glycols and Compounds Related to Sugars 




O— O 



(CHsl^N 



Mycaminose 
(carbomycins A, B) 




CH3O 



H2N— C— O OH 

Noviose 
(novobiocin) 




O— (CHalzN 



HO 

My CO rose 

(carbomycins A, B, 

spiramycins) 

HOCH 




O— 



O— 



2,3,4,6-Tetradeoxy-4- 

dimethylaminohexopy- 

ronose (spiramycins) 




O— 



(CH3)2N OH 

Amosamine 
(amice tin) 




OH 

L-2-Ketofucopyranose 
(angustmycin A) 



CH2OH 





OH OH 

5-Keto-6-deoxy- D-Talose Neoinosamine-2 

arabohexose (hygromycin B) (hygromycin A) 

(hygromycin A) 

(Two hydroxyl groups in neoinosamine-2 of hygromycin A are connected 
in a methylenedioxy bridge. Homomycin contains a similar sugar.) 



HOCH 





CH2OH 



NH OH 



HOCH2 




3-Deoxy-3-amino-D- 

ribose 

(puromycin) 



Cordycepose 
(cordycepin) 



a-D-Gulosamine 

(streptothricin, 

roseothricin) 



Pfizer Handbook of Microbial Metabolites 



22 




CH20CH3 



CH— N= 

I 
CHOH 



CHj 



CH3 
Rhodosamine 
(rhodomycin) 



Methyl-2,4-dideoxy-2-aminotetroside 
(elaiomycin) 



Good reviews of aminosugars have been published. ^^'^"^ 

Other unusual sugars have been identified as components of 
the polysaccharides, mucopolysaccharides, etc., which occur in 
microbial cell walls and other cell tissues. Information can be 
obtained on these by way of Appendix A. 

No attempt will be made here to discuss thoroughly the poly- 
saccharides. Many references to this subject are listed in Ap- 
pendix A and in the Bibliography. 

As mentioned above many of the large molecules of micro- 
organisms are mucopolysaccharides, etc., which contain sugars 
other than glucose. Glucose is in fact a relatively rare com- 
ponent of such molecules, but galactose, galacturonic acid, 
fucose, mannose and other sugars are common. Many hexoses 
and pentoses can be formed from the parent sugar without 
chain rupture. The intermediates in these interconversions are 
known to be sugar nucleotides:^' 



X Sugar 



Base (Uracil 
or 
Guanine) 




OH OH 



I > > Y Sugar 

Epimerization 
Oxidation 
Reduction 
Decarboxylation 



i^'T. Naito, Jap. }. Pharm. and Chem. 31 23 (1959). 

^''' A. B. Foster and D. Horton, "Advances in Carbohydrate Chem- 



23 Alcohols, Glycols and Compounds Related to Sugars 

Some of these reactions are reversible. Some of the less com- 
mon aminohexoses are formed also in this way from glucosa- 
mine. 

Certain fatty alcohols are classified in this chapter because 
of their functional groups, although biosynthetically they are 
more compatible with the fatty acids. 

15 Ethanol, C^H^O, colorless liquid, b.p. 78.5°, n^'" 1.3610. 

CHsCHoOH 

Yeasts, fusaria, mucors, penicillia, aspergilli, etc. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
1954 Vol. I pp. 17-196. 

16 Dihydroxyacetone, C;^H,jO;5, colorless microcrystalline powder, 

m.p. 75-80° (polymorphic). 

HOCH,— C— CH2OH 



Acetobacter suboxydans (on glycerol) 

Aurel Puskas, Yearbook Inst. Agr. Cheni. Technol. Univ. 
Tech. Sci. Budapest, Hung. 3 (1952). 

Idem., ibid. 8 150 (1954). 

A 90% yield of crude and a 70% recovery on recrystal- 
lization was reported. 

Dihydroxyacetone has been reported also in cultures 
of Penicillium brevi-compactum and Corynebacterium 
diphtheriae (on glucose). 

Michizo Asano and Hideo Takahashi, /. Pharm. Soc. Japan 
68 186 (1948); Paul Godin, Biochim. et Biophys. Acta 11 
114 (1953). 

17 Glycerol (Glycerin, 1,2,3-Propanetriol), C3H8O3, m.p. 17.8°, b.p. 
290° (dec), n,r" 1.4746. 

CH.2— CH— CH2 

i I I 

OH OH OH 

Yeasts, Bacillus subtilis, Aspergillus wentii, Clastero- 
sporia, Helminthosporia, penicillia, etc. 

Numerous recent patents. The glycerol situation is well 
summarized in Underkofler and Hickey, "Industrial Fermenta- 
tions," Chemical Publishing Co., Inc., New York, N. Y., 1954 
Vol. I; L. A. Underkofler, Glycerol, chap. 8, pp. 252-270. 

istry," Aspects of the chemistry of the amino sugars. Academic Press, 
New York, N. Y., 1959 Vol. 14 pp. 224-233. 

^' Saul Roseman, Federation Proc. 18 984 (1959). (A review) 



Pfizer Handbook of Microbial Metabolites 24 

18 n-Butanol, C4H10O, colorless liquid, b.p. 117°, n^^" 1.3993. 

CH3CH2CH2CH2OH 

Clostridium acetobutylicum, CI. propylbutylicum, 
CI. saccharobutylicum 

Yields of about 30% mixed solvents, mainly butanol, 
but containing also acetone, isopropanol and ethanol are 
common. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. I; W. N. McCutchan and R. J. Hickey, The 
butanol-acetone fermentations, chap. 11, pp. 347-388. 

19 2,3-Butanediol, C4H10O0, colorless liquid, b.p. 180°. 

The optical isomer produced depends on the micro- 
organism. 

CH3— CH— CH— CH3 

I I 
OH OH 

Aerobacter aerogenes, Serratia marcescens. Bacillus 
polymyxa, Bacillus subtilis, Pseudomonas hydrophila, Ba- 
cillus mesentericus, yeasts 

Acetoin, diacetyl and alcohol are often produced at the 
same time. Approximately 90% yields of butanediol have 
been reported. 

J. A. Wheat, Ind. Eng. Chem. 45 2387 (1953). 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. II; G. A. Ledingham and A. C. Neish, Fer- 
mentative production of 2,3-butanediol, chap. 2, pp. 27-93. 

Heikki Suomalainen and Lauri Jannes, Nature 157 336 
(1946). 



20 Erythritol, C4H10O4. 



CH2— CH— CH— CH2 
OH OH OH OH 



Armillaria mellea 

J. H. Birkinshaw, C. E. Stickings and P. Tessier, Bio- 
chem. }. 42 329-332 (1948). 

Thirteen % of dry myceUum was the D-threitol isomer, 
colorless needles, m.p. 88.5°, [aW +4.3° (c 1 in water), 
— 11.1° (in 95% ethanol). Other isomers have been re- 



25 Alcohols, Glycols and Compounds Related to Sugars 

ported, especially z-erythritol (meso-erythritol). Colorless 
prisms, m.p. 120° (121.5°) from: 

Roccella montagnei (yield 2% ) and other Roccella spe- 
cies, Penicillium brevi-compactum, P. cyclopium, Asper- 
gillus terreus, etc. 

Albert E. Oxford and Harold Raistrick, Biochem. }. 29 1599 
(1935). 

Yosio Sakurai, /. Pharm. Soc. Japan 61 108 (1941). 

21 D-Lyxuronic Acid (isolated as the calcium salt) C-.HjOgCa/ 
2-2HoO, [aW -23° 30 minutes -53° (in water). 



COOH 
HOCH 
HCOH 
HCOH 
CHO 

Acetobacter melanogenum 

Minoru Ameyama and Keiji Kondo, Bull. Agr. Chem. Soc. 
(Japan) 22 271 (1958). 

22 d-Arabitol, C-HioOr,, colorless spheroid crystals, m.p. 103°, [ajn^" 
+7.7° (c 9.26 in saturated borax solution). 

CH2— CH— CH— CH— CH2 

I I I I I 

OH OH OH OH OH 

Lobaria pulmonaria Hoffm., Ramalina geniculata Tayl., 
R. sinensis, R. tayloriana, R. scopulorum (Retz. ) Nyl., 
Cladonia impexa Harm., Fistulina hepatica, Lecanora 
gangaleoides, Parmelia latissima Fee, Umbilicaria pustu- 
lata 

Yasuhiko Asahina and Masaichi Yanaglta, Ber. 67B 799 
(1934). 

T. W. Breaden, J. Keane and T. J. Nolan, Sci. Proc. Roy. 
Dublin Soc. 23 6 (1942). 

Yngve Johannes Solberg, Acta Chem. Scand. 9 1234 (1955). 



Pfizer Handbook of Microbial Metabolites 26 

23 2,5-Diketogluconic Acid, CgHgOT, isolated as Ca salt. No good 

m.p. 

COOH 

I 

c=o 

HOCH 
HCOH 

c=o 

I 
CH.OH 

Acetobacter melanogenum, Pseudomonas, Phytomonas 
spp. 

H. Katznelson, S. W. Tanenbaum and E. L. Tatum, J. Biol. 
Chem. 204 43 (1953). 

24 Glucosone, CgHioOe, levorotatory syrup with reducing properties. 

CHO 

c=o 

I 
HOCH 

HCOH 

HCOH 

CHoOH 

Aspergillus parasiticus, A. fiavus, some algae 
Yields of 13-17% from sucrose have been reported. 
Cecil R. Bond, Edwin C. Knight and Thomas K. Walker, 
Biochem. J. 31 1033 (1937). 

Ross C. Be_an and W. Z. Hassid, Science 124 171 (1956). 

25 2-Ketogluconic Acid, C^HioO^, colorless crystals, m.p. 152° (Me 

ester, m.p. 172°). 

COOH 

c=o 

1 
HOCH 

HCOH 

HCOH 

CH2OH 
Acetobacter melanogenum, Pseudomonas, Phytomonas 
spp. 



27 Alcohols, Glycols and Compounds Related to Sugars 

The yields of 2-ketogluconic acid are better than 70%. 
2,5-Diketogkiconic acid can be made the principal prod- 
uct. This diketo acid is unstable, but can be isolated as a 
salt. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Cliemical Publishing Co., Inc., New York, 
1954 Vol. II, Lewis B. Lockwood, Ketogenic fermentation 
processes, chap. 1, pp. 13-14. 

H. Katznelson, S. W. Tanenbaum and E. L. Tatum, /. Biol. 
Chem. 204 43 (1953). 

26 5-Ketogluconic Acid, CeHmOj, generally isolated as the Ca salt 

(no sharp m.p.). 

COOH 
HCOH 
HOCH 
HCOH 

c=o 

CH>OH 

Acetobacter suboxydans 

Yields of about 90% have been reported. 

Shiro Teramato, Riichiro Yagi and Ichiro Hori, /. Fermenta- 
tion Technol. (Japan) 24 22 (1946). 

Joseph J. Stubbs, Lewis B. Lockwood, Edward T. Roe and 
George E. Ward, U. S. Patent 2,318,641 (1943). 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. II, Lewis B. Lockwood, Ketogenic fermenta- 
tion processes, chap. 1, pp. 10-12. 

27 2-Ketogalactonic Acid, C.-HioO^, colorless crystals, m.p. 170° (K 

salt, m.p. 139°; Me ester, m.p. 138°). 

COOH 

c=o 

HOCH 
HOCH 
HCOH 
CHoOH 



Pfizer Handbook of Microbial Metabolites 28 

Pseudomonas species (on galactose) 
Toshinohu Asai, Ko Aida and Yashuiro Ueno, /. Agr. Chem. 
Soc. Japan 25 625 (1951-1952). 

28 D-GIucuronic Acid, C6H10O7, colorless needles, m.p. 165°, [ccW* 

+ 11.7°^ +36.3° (2 hours, c 1 in water). 

CHO 

I 
HCOH 

I 
HOCH 

HCOH 

I 
HCOH 

I 
COOH 

Ustulina vulgaricus 

H. Wunchendoroff and C. Killian, Compt. rend. 187 
572 (1928). 

Not isolated — manner of identification not mentioned. 

Penicillium sp. 

Gizin Itto, /. Agr. Chem. Soc. Japan 9 552 (1933). 

K. Sivarama Sastry and P. S. Sarma, Nature 179 44 (1957). 

29 Saccharic Acid, CeHnjOg, colorless needles, m.p. 125°, [aW^ 

+6.86° -^ 20.6° (c 1 in water). 

COOH 

HCOH 

I 
HOCH 

I 
HCOH 

I 
HCOH 

I 
COOH 

Aspergillus niger 

T. K. Walker, Vira Subramanian and Frederick Chal- 
lenger, J. Chem. Soc, 3044 (1927). 

About 3.6 g. of the potassium salt were obtained from 
120 g. of glucose by interrupting the fermentation before 
the appearance of much citric or oxalic acids. Also fer- 
mentation of 20 g. of calcium gluconate gave 3.7 g. of 
calcium saccharate. 



29 Alcohols, Glycols and Compounds Related to Sugars 

Also reported formed from glucose by two yeasts, An- 
thomijces renkaufi and Amphierna rubra: 
J. Gruss, Jahrb. luiss. Botan. 66 109 (1926). 

30 meso-Inositol, C^HjoOo (dihydrate), colorless crystals, m.p. 218° 
(anhydrous) 250-253°. 




H OH 



Pseudomonas fluorescens, Serratia marcescens, Proteus 
vulgaris, Clostridium butylicum, yeasts 

Yields of 2700-5000 /xg. per gram of dry cell weight are 
obtained in brewers' yeast. 

Inositol Literature Briefs Tech. Bull. Y3-101, Corn Products 
Refining Co., 1953, 44 pp. (A bibliography with titles and 
abstracts) 

31 D-Gluconic Acid, CgHioO^, colorless syrup, cannot be isolated, but 
readily forms (principally) the 8-lactone, white crystals, 
m.p. 153°, [aU +63.5° -^ +6.2° (elm water). 

COOH 

HCOH 

HOCH 

I 
HCOH 

I 
HCOH 

CH2OH 

Wide variety of mold species, acetobacter species, etc. 

Yields 95% with Aspergillus niger. 

A. J. Mayer, E. J. Umberger and J. J. Stubbs, Ind. Eng. 
Chem. 32 1379 (1940). 

Leland A. Underkofier and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. I, L. A. Underkofier, Gluconic acid, chap. 14, 
pp. 446-469. 



Pfizer Handbook of Microbial Metabolites 30 

32 D-Mannonic Acid, CfjHioO^, forms y- or 8-lactones, but the free 
acid cannot be isolated pure. 



COOH 

J 

HCOH 

I 
HCOH 

HOCH 

HOCH 

CH,OH 

P. purpurogeninn var. rubrisclerotium (on D-mannose) 
Acetobacters 

Galactonic acid, etc., can be produced similarly from 
the corresponding sugar. 

A. Angeletti and C. F. Cerruti, Ann. chim. applicata 20 424 
(1930). 

33 D-Glucosamine (Chitosamine) CeHigOr.N, white needles, m.p. 
110° (dec), [a],;-"' +47.5° (c 1 in water). 

CHO 

I 
HC— NH2 

I 
HOCH 

HCOH 

HCOH 

CH.OH 

Many bacteria, fungi and lichens. Present in bound 
form in mold mycelium. Produced by the action of cer- 
tain streptomyces species on chitin. 

Joseph J. Noval and Walter J. Nickerson, Bacteriol. Proc, 
125 (1956). 

LesUe Ralph Berger and Donald M. Reynolds, Biochim. et 
Biophys. Acta 29 522 (1958). 



31 Alcohols, Glycols and Compounds Related to Sugars 

34 Mesoinositol Monophosphate, C.jHisOgP-SH^O, colorless tablets, 
m.p. 201° (dec.)- 

OH 

O— PO(OH)2 




Mycobacterium tuberculosis var. hominis 

Michael A. Macheboeuf, Georgette Levy and Marguerite 

Faure, Compt. rend. 204 1843 (1937). (Occurred as a fatty 

acid ester) 

James Cason and R. J. Anderson, J. Biol. Chem. 126 527 

(1938). (As a constituent of a polysaccharide) 

35 D-Mannitol, CgHijOe, colorless prisms, m.p. 163° (166°) [aln"^ 

—0.49°. (C 1 in water. Addition of borax ^^ strong dex- 
trorotation. ) 

CH2— CH— CH— CH— CH— CH. 

OH OH OH OH OH OH 

Aspergilli, penicillia, other fungi, many lichens, algae 
and bacteria 

For example: Mitizo Asano, Chunoshin Ukita and 
Tomoyoshi Komai, Japanese Patent 180,442 (1949) describe 
extraction of mannitol and ergosterol from Penicillium 
mycelium. See W. Karrer's compilation (listed in the general 
reference bibliography) for other references. 

36 D-Volemitol, C^H^-O-, silky needles, m.p. 153.5° [a],r" +17.08° 

(1.001 g. + 0.7 g. of Borax in 15 ml. of H.O). 

CHo— CH— CH— CH— CH— CH— CH2 
OH OH OH OH OH OH OH 

Dermatocarpon miniatum (L.) Mann. 

Yasuhiko Asahina and Motoyasu Kagitani, Ber. 67B 804 
(1934). 

Bengt Lindberg, Alfons Misiorny and Carl Axel Wacht- 
meister. Acta Chem. Scand. 7 591 (1953). (A survey of the 
occurrence of low molecular weight carbohydrate constituents 
in lichens ) 



Pfizer Handbook of Microbial Metabolites 



32 



37 6-O-Acetylglucose, C8H14O7, minute colorless prisms, m.p, 133°, 

[(x]d^° +48° (c 4.0 in water at equilibrium). 

CHO 

I 
HCOH 

1 
HOCH 

HCOH 

I 
HCOH 

I 
CH2OCOCH3 

Bacillus megaterium 

R. B. Dufe, D. M. Webley and V. C. Farmer, Nature 179 
103 (1957). 

38 D-Mannopyranosyl-1-meso-erythritol, C10H20O9, colorless crystals, 

m.p. 160°, [a]v -36.7°. 



CH2OH 




-CH2 



-CH— CH— CH2 

1 I I 
OH OH OH 



Ustilago sp. 

Besides this water-soluble compound the fungus pro- 
duces 15 g. per liter of an oil, consisting of a mixture of 
fatty acid esters of o-mannopyranosyl-l-meso-erythritol. 

B. Boothroyd, J. A. Thorn and R. H. Haskins, Can. J. 
Biochem. and Physiol. 34 10 (1956). 

39 Umbilicin (3-yg-D-Galactopyranosido-D-arabitol), C11H22O10, color- 
less crystals, m.p. 138°, [a]D"° —81° (c 2 in water). 

CH2OH 



HOCH 

I 
HC— 



-0— CH 



HCOH HCOH 

I I 

CH2OH HOCH 

I 
HOCH 

I 
HC 



CH,OH 



33 



Alcohols, Glycols and Compounds Related to Sugars 



Umbilicaria pustidata 

Bengt Lindberg, Carl A. Wachtmeister and Borje Wickberg, 
Acta Chem. Scand. 6 1052 (1952). 

40 Trehalosamine, CjoHooOioN (Hydrochloride) white microcrys- 
talMne powder, [a] d'' +176° (c 2.0 in water). 




CH2OH 



A streptomycete 

A yield of about 5 g. per liter was obtained. 
Frederico Arcamone and Franco Bizioli, Gazz. chim. ital. 
87 896 (1957). 

41 Leucrose (5-0-a-D-Glucopyranosyl-D-fructopyranose), C12H22O11, 
colorless hygroscopic bars, m.p. 161-163° (anhydrous), 



156-158° (monohydrate), [a]^'' -8.2' 
c 4 in water). 



CH2OH 




■7.6° (<lhour, 



CH2OH 



Leuconostoc mesenteroides 

Frank H. Stodola, E. S. Sharpe and H. J. Koepsell, J. Am. 
Chem. Soc. 78 2514 (1956). 

42 Kojibiose ( 2-0-a-D-Glucopyranosyl-D-glucose ) , CioHooOn, m.p. 
(Octaacetate) 166°, [aU +150° (c 2.1 in chloroform). 
Free sugar: [aU +136° (equil., c 0.5 in water). 




CH2OH 



Pfizer Handbook of Microbial Metabolites 



34 



Aspergillus niger 

Stanley Peat, W. J. Whelan and Kathleen A. Hinson, Chem. 
and Ind., 385 (1955). 

A. Sato and K. Aso, Nature 180 984 (1957). 

43 Trehalose (Mycose, a-D-Glucosido-a-D-glucoside), CioHooOu, 
colorless, hygroscopic crystals, m.p. ~210° (dec.) (anhy- 
drous), 97° (hydrate), [<x]t>-° (hydrate) +178° (in water). 




CH2OH 



Amanita muscaria, other mushrooms and molds, myco- 
bacteria, yeasts and algae. First isolated from rye ergot 
(Claviceps purpurea (Fr. ) Tul.). 

Trehalose is present in young mushrooms, but as the 
plants develop it is replaced by mannitol. It also occurs 
in seaweeds and higher plants. 

E. Bourquelot, Compt. rend. 108 568 (1889). 

H. Bredereck, Ber. 63B 959 (1930). (Structure) 

Bengt Lindberg, Acta Chem. Scand. 9 917 (1955). 

44 Lactobionic Acid, CioHooOja, Calcium Salt : granular white pow- 
der, [a],r' +25.1° (c 5.2 in water). 



COOH 

HCOH 

- I 
HOCH 

HC 



CH— 

I 
HCOH 

I 

HOCH 
HOCH 
HC 



HCOH 

I I 

CH2OH CH2OH 

Pseudomonas species, other oxidative bacteria (on lac- 
tose) 

A 77 '^f yield has been reported. Maltobionic acid was 
prepared similarly from maltose. 

Frank H. Stodola and Lewis B. Lockwood, J. Biol. Chem. 
171 213 (1947). 



35 Alcohols, Glycols and Compounds Related to Sugars 

Lewis B. Lockwood and Frank H. Stodola, U. S. Patent 

2,496,297(1950). 

45 Hygromycin B, Ci.-.HosOioNo, amorphous powder, m.p. ^-'180°. 

D-Talose has been shown to be one moiety of this anti- 
biotic. 

Streptomyces hygroscopicus 

Robert L. Mann and W. W. Bromer, J. Am. Chem. Soc. 80 
2715 (1958). 

Paul F. Wiley and Max V. Sigal, Jr., ibid. 80 1010 (1958). 

46 Grifolin, CieHo^Oo, fine colorless needles, m.p. 40°. 

CHa OH 

\ I 

C=CH— CH.— CHj— C=CH— CH=CH— CH— C— CH.— CHs 

/ I II 

CHs CH3 OH CH3 

Grifola confluens (=Polyporus confLuens) 

Other components of the extract were mannitol, sterols, 

a hemin-like substance, a compound C.SH14O (m.p. 145°) 

and a compound Ci,r,H^,402 (m.p. 151°). 

Y. Hirata and K. Nakanishi, /. Biol. Chem. 184 135 (1950). 

47 Cetyl Alcohol, C16H34O, colorless crystals, m.p. 50°, Ud'^ 1.4283. 

CHsICHzIhCHsOH 

Amanita phalloides 

Heinrich Wieland and Gustav Coutelle, Ann. 548 270 
(1941). 

48 Clavicepsin, CisH^^Oip,, colorless crystals, m.p. (anhydr. ) 198°, 

[aln-" +142°. 

A glucoside hydrolyzing to 1 mole of mannitol and 2 
moles of glucose. The detailed structure was not deter- 
mined. 

Claviceps purpurea 

F. Marino-Zuco and U. Pasquero, Gazz. chim. ital. 41 368 
(1912). 

49 Stearyl Alcohol, CigHayO, colorless leaflets, m.p. 59°. 

CHslCHslifiCHoOH 

Penicillium notatum 

A yield of 0.13 g. was obtained from 300 g. of dry 
mycelium. 



Pfizer Handbook of Microbial Metabolites 



36 



A. Angeletti, G. Tappi and G. Biglino, Ann. chim. (Rome) 
42 502 (1952). 

50 d-2-Octadecanol, CigHgsO, colorless needles, m.p. 56°, [ctjo +5.7° 

(in chloroform). 

CHslCHalisCHCHa 

I 
OH 

Mycobacterium tuberculosis var. hominis, M. avium, 
M. phlei 

Mary C. Panghorn and R. J. Anderson, /. Am. Chem. Soc. 
58 10 (1936). 

R. E. Reeves and R. J. Anderson, ibid. 59 858 (1937). 

R. J. Anderson, J. A. Crowder, M. S. Newman and F. H. 
Stodola, J. Biol. Chem. 113 637 (1936). 

51 d-3-Octadecanol, CisHggO, colorless crystals, m.p. 56°. 

CH3(CH2),4CHCH2CH3 

OH 

Corynebacterium diphtheriae 

A. A. Kanchukh, Ukrain. Biokhim. Zhur. 26 186 (1954). 

52 Kanamycin, C18H36O11N4, Sulfate: white prisms which decom- 

pose over a wide range above 250°, [ajn"* +146° (c 1 in 
0.1 N sulfuric acid). 



CH2NH 



6-Glucosamine< 



Kanosamine 




>2-Deoxystrep- 
tamine 



Streytomyces kanainyceticus 

Tomio Takeuchi, Tokuro Hikiji, Kazuo Nitta, Seiro Yama- 
zaki, Sadao Abe, Hisaro Takayama and Hamao Umezawa, /. 
Antibiotics (Japan) lOA 107 (1957). 

Hamao Umezawa, Mashiro Ueda, Kenji Maeda, Koki 



37 Alcohols, Glycols and Compounds Related to Sugars 

Yagishita, Shinichi Kondo, Yoshiro Okami, Ryozo Utahara, 
Yasuke Osato, Kazuo Nitta and Tomio Takeuchi, ibid. lOA 
181 (1957). 

Kenji Maeda, Masahiro Ueda, Koki Yagishita, Shohei 
Kawaji, Shinichi Kondo, Masao Murase, Tomio Takeuchi, 
Yoshiro Okami and Hamao Umezawa, ibid. lOA 228 (1957). 

M. J. Cron, D. L. Johnson, F. M. Palermiti, Y. Perron, H. D. 
Taylor, D. F. Whitehead and I. R. Hooper, }. Am. Chem. Soc. 
80 752 (1958). 

M. J. Cron, O. B. Fardig, D. L. Johnson, D. F. Whitehead, 
I. R. Hooper and R. U. Lemieux, ibid. 80 4115 (1958). 

53 Kanamycin B, colorless crystals, m.p. dec. from 170°, [ajn^* 

+ 135° (c 0.63 in water). 

Acid hydrolysis yields 2-deoxystreptamine and kanosa- 
mine, but no 6-glucosamine as from kanamycin. An un- 
identified ninhydrin-positive compound was obtained in- 
stead. Positive Schiff, Molisch, Elson-Morgan tests. 

Streptomyces kanamyceticiis 

H. Schmitz, O. B. Fardig, F. A. O'Herron, M. A. Rousche 
and I. R. Hooper, /. Am. Chem. Soc. 80 2911 (1958). 

54 Streptomycin, C01H39O12N7, m.p. (Reineckate) 164° dec. (He- 

lianthate) 220-226° dec, [ajc (Hydrochloride) -84° (c 
0.5 in water), [aU^"-^ (Trihydrochloride) -86.1° (c 1.0 
in water), [a]u'^ (Sulfate) -79° (c 1.0 in water). Salts 
are deliquescent. 



NH— C— NH2 



H2N— C— NH I 



Streptobiosamine 




Streptidine 



Streptose 



N-Methyl-L-glucosamine 



Pfizer Handbook of Microbial Metabolites 



38 



Streptomyces griseus (Krainsky) Waksman et Henrici 
S. bikiniensis, S. mashuensis 

Albert Schatz, Elizabeth Bugie and Selman A. Waksman, 
Proc. Soc. Exptl. Biol. Med. 55 66 (1944). (Isolation) 

Selman A. Waksman, "Streptomycin, Its Nature and Appli- 
cations," Williams and Wilkins Co., Baltimore, Md., 1949. (A 
review ) 

Herbert E. Carter, R. K. Clark, Jr., S. R. Dickman, Y. H. 
Loo, P. S. Skell and W. A. Strong, J. Biol. Chem. 160 337 
(1945). 

Frederick A. Kuehl, Jr., Robert L. Peck, Charles E. Hoffhine, 
Jr., Robert P. Graber and Karl Folkers, J. Am. Chem. Soc. 68 
1460 (1946). 

Frederick A. Kuehl, Jr., Edwin H. Flynn, Norman G. Brink 
and Karl Folkers, ibid. 68 2096, 2679 (1946). (Structure) 

I. R. Hooper, L. H. Klemm, W. J. Polglase and M. L. 
Wolfrom, ibid. 69 1052 (1947). 

H. E. Carter, R. K. Clark, Jr., S. R. Dickman, Y. H. Loo, 
P. S. Skell and W. A. Strong, Science 103 540 (1946). 

E. P. Abraham and H. W. Florey, "Antibiotics," Oxford Uni- 
versity Press, London, 1949 Vol. II chap. 41, pp. 1297-1309. 

E. P. Abraham, ibid. chap. 42, pp. 1310-1326. 

55 Hydroxystreptomycin (Reticulin) C2iH;j,,05:iN7, Helianthate: red- 
brown crystals, m.p. : darkening at 200° (dec.), Trihydro- 
chloride: [a]n-^ -91° (c 1.0 in water). 



NH— C— NH, 




39 Alcohols, Glycols and Compounds Related to Sugars 

Streptomyces griseocarneus 

Seigo Hosaya, Momoe Soeda, Nobuhiko Komatsu and Yoko 
Sonoda, Japan. J. Exptl. Med. 20 327 (1949). 

Robert G. Benedict, Frank H. Stodola, Odette L. Shotwell, 
Anne Marie Borud and Lloyd A. Lindenfelser, Science 112 77 
(1950). 

Frank H. Stodola, Odette L. Shotwell, Anne Marie Borud, 
Robert G. Benedict and Arthur C. Riley, Jr., /. Am. Chem. Soc. 
73 2290 (1951). ( Structure ) 

56 Dihydrostreptomycin, C^iH4iOi2N7, non-deliquescent white pow- 
der [a]D"' —94.5°. Hydrochloride and sulfate were used. 



NH— C— NH2 



H2N— C— NH 




Streptomyces humidus 

Sueo Tatsuoka, Tsunaharu Kusaka, Akira Miyake, Mi- 
chitaka Inoue, Hiromu Hitomi, Yutaka Shiraishi, Hidesuke 
Iwasaki and Masahiko Imanishi, Pharm. Bull. 5 343 (1957). 
(Primary fermentation product) 



Pfizer Handbook of Microbial Metabolites 



40 



57 Hygromycin A, C23H29O12N, amorphous (some crystalline deriva- 
tives have been prepared), [aln"'^ —126° (c 1 in water). 
Partial structure: 



HO- 

O 

1 
HC- 



/ \— CH— C— C— NH 
^ CH3 



HOCH 

HCOH 

I 
HC 




c=o 



CH3 



Streptomyces hygroscopicus (Jensen) Waksman and 
Henrici 

R. L. Mann, R. M. Gale and F. R. Van Abeele, Antibiotics 
and Chemotherapy 3 1279 (1953). (Isolation) 

Robert L. Mann and D. O. Wolf, J. Am. Chem. Soc. 79 120 
(1957). (Structure) 

58 Homomycin, white powder, m.p. 105-109° (dec. >160°). 

Homomycin has been shown to be the same as hygro- 
mycin except that the homomycin amino sugar moiety is : 





Streptomyces noboritoensis n. sp. 

Yusuke Sumiki, Gotaku Nakamura, Makoto Kawasaki, 
Satoru Yamashita, Kentaro Anzai, Kiyoshi Isono, Yoshiko 
Serizawa, Yoko Tomiyama and Saburo Suzuki, J. Antibiotics 
(Japan) 8A 170 (1955). (Isolation) 

Mitsuo Namiki, Kiyoshi Isono, Kentaro Anzai and Saburo 
Suzuki, ibid. lOA 36 (1957). (Structure) 



41 



Alcohols, Glycols and Compounds Related to Sugars 



59 Paromomycin, C23H45O14N5, white amorphous solid, [ajn^^ +64° 
(c 1.0 in water). Hydrochloride [ix]d~'' +56.5° (c 1.0 in 
water). 



2-Deoxystreptamme 




Paromamine 



Paromobiosamine 



D-Ribose 



Streptomyces rimosus forma paromomycinus 
Paromomycin seems to be identical with catenuHn. 
Theodore H. Haskell, James C. French and Quentin R. Bartz, 
J. Am. Chem. Soc. 81 3480 (1959). 
Ibid. Belgian Patent 547,976. 

60 Neomycins. (Fradiomycins, Streptothricins, Neomins, Mycifra- 
din, Nivemycins, Myacins) 

Neomycin A is identical with neamine, a moiety of 
neomycins B and C. Neomycins B and C are identical ex- 
cept for the diaminohexose components. 

Neomycin B ( Streptothricin B II), C23H46O12N6, amor- 
phous hygroscopic white powder, no definite m.p., [ajn'^ 
+83° (in 0.2 N H0SO4). 

C12H05O5N4— O— CsHeOlOHla— O— C6H70(OH)2(NH2)2 



Neamine 
(structure 
unknown) 



D-Ribose 



Diaminohexose B 
(structure unknown) 



Neomycin C (Streptothricin B I), C23H46O12N6, amor- 
phous, hygroscopic white powder, no definite m.p., [ajn""' 
+ 121° (in 0.2 N H2SO4). 



Pfizer Handbook of Microbial Metabolites 42 

Also contains neamine. The disaccharide portion 
(neobiosamine C) has been characterized, however, as: 



CH2NH2 




D-Ribose Neosamine C 

Streptomyces fradiae, other Streptomyces spp. 

Selman A. Waksman and Hubert A. Lechevalier, Science 
109 305 (1949). (Isolation) 

Byron E. Leach, William H. DeVries, Harrison A. Nelson, 
William G. Jackson and John S. Evans, J. Am. Chem. Soc. 73 
2797 (1951). (Isolation) 

Jared H. Ford, Malcolm E. Bergy, A. A. Brooks, Edward R. 
Garrett, Joseph Alberti, John R. Dyer and H. E. Carter, ibid. 
77 5311 (1955). 

Kenneth L. Rinehart, Jr., Peter W. K. Woo, Alexander D. 
Argoudelis and Astrea M. Giesbrecht, ibid. 79 4567 (1957). 

Kenneth L. Rinehart, Jr., Peter W. K. Woo and Alexander D. 
Argoudelis, ibid. 79 4568 (1957). 

Idem., ibid. 80 6461 (1958). 

Kenneth L. Rinehart, Jr., and Peter W. K. Woo, ibid. 80 
6463 (1958). (Structure) 

61 Catenulin (Sulfate) [a]ir^ +51.9° (c 1 in water). 

A substance resembling paromomycin. Acid hydrolysis 
yields neamine.* 

Streptomyces catenulensis 

J. W. Davisson, I. A. Solomons and T. M. Lees, Antibiotics 
and Chemotherapy 2 460 (1952). 

62 Dextromycin, Helianthate: m.p. 227°, Hydrochloride: [a]u'^ 

+61° (c 1 acetone). 

Similar to neomycin B.* 
Streptomyces sp. resembling S. fradiae 
Koichi Ogata and Koichi Nakazawa, J. Antibiotics (Japan) 
3 440 (1950). 
* Probably identical with paromomycin. (Private communication 
from Drs. W. Celmer and C. Shaffner) 



43 Alcohols, Glycols and Compounds Related to Sugars 

Toyonari Araki, Akira Miyake, Yoshitamo Aramaki, Hiroshi 
Kojima, Hajime Yokotani, Koichi Ogata and Koichi Nakazawa, 
Ann. Repts. Takeda Research Lab. 13 1 (1954). 

* Identical with neomycin B. See addendum. 

63 Framycetin (Actilin, Soframycin, Antibiotic E.F. 185), Hydro- 

chloride: white powder, [a]i. +57° (c 1.0 in water), m.p. 
(picrate) 189° (dec). 

Framycetin resembles neomycin and streptomycin in 
some respects, but is distinct. Hydrolysis yields neamine, 
a pentose, and a diaminohexose. Framycetin forms pep- 
tide derivatives such as a reineckate and a picrate. The 
molecular weight is about 1400-1500. No guanidine 
tests were observed, and all the nitrogen is present as pri- 
mary amine groups. 

Streptomyces sp. resembling S. lavendulae 

Louis Jacques Decaris, Ann. pharm. frang. II 44 (1953). 

Maurice Marie Janot, Henry Penau, Digna van Stolk, Guy 
Hagemann and Lucien Penasse, Bull. Soc. chim. France, 1458 
(1954). 

A. Lutz and M. A. Witz, Compt. rend. soc. biol. 149 1467 
(1955). 

A. Saito and C. P. Schaffner, Congr. intern, biochim.. 
Resumes communs., 3"^ Congr., Brussels, 1955, p. 98. 

64 Hydroxymycin, probable empirical formula C._,r,H470i5N-,, white 

powder, [a]ir" 63° ± 2° (c. 1.0 in water) (Sulfate) white 
powder, [aic-" +51° (c 1.0 in water). 

A basic antibiotic similar to streptomycin and neomy- 
cin. Contains 6.2% total nitrogen and 6.0% amino nitro- 
gen. It is water soluble and insoluble in most organic 
solvents with a molecular weight of about 610. Hydroly- 
sis yields a fragment called pseudoneamine and others 
which show pentose and 2-aminohexose reactions. 

An antifungal substance was produced in the same 
culture. 

Streptomyces paucisporogenes 

M. M. Janot, H. Penau, G. Hagemann, H. Velu, J. Teillon 
and G. Bouet, Ann. pharm. frang. 12 440 (1954). 

G. Hagemann, G. Nomine and L. Penasse, Ann. pharm. 
frang. 16 585 (1958). 

H. Penau, G. Hagemann and H. Velu, Bull. soc. chim. biol. 
41 761 (1959). 

J. Bartos, Ann. pharm. frang. 16 596 (1958). 



Pfizer Handbook of Microbial Metabolites 



44 



65 Mannosidostreptomycin (Streptomycin B), C27H49O17N7, color- 
less crystals, m.p. (Anhydrous Reineckate) 178° dec. 
(Trihydrochlorlde) 190-200° dec, [a]u~^ (Trlhydrochlo- 
ride) -47° (c 1.35 in water). 



CH2OH 



NH NH 

II II 

H2N— C— NH NH— C— NH2 




O^ 




Occurs together with streptomycin in some cultures. 
Streptomyces griseus 

Josef Fried and Homer E. Stavely, /. Am. Chem. Soc. 74 
5461 (1952). (Structure) 

66 Phthiocerol, C36H74O3, colorless plates, m.p. 71.5-73°, [(x]j> 
-4.50° (c 11.48 in chloroform). 



45 Alcohols, Glycols and Compounds Related to Sugars 

It is claimed (in the most recent reference below) that 
phthiocerol, as ordinarily isolated, is a mixture of the fol- 
lowing two substances: 

OCH3 

I 
CH3(CHo)ooCH— CHo— CH{CH2)4CH— CH— CH2CH3 

OH OH CH3 



and 



OCH3 

CHslCHsl^oCH— CH2— CH(CH2)4— CH— CH— CH2CH3 

I I I 

OH OH CH3 

Mycobacterium tuberculosis (human, bovine and 
avian ) 

In the wax of the mycobacteria phthiocerol is present 
mainly as the dimycoceranate. 

J. A. Hall, J. W. Lewis and N. Polgar, /. Chem. Soc, 3971 
(1955). 

Hans NoU, /. Biol. Chem. 224 149 (1957). 

H. Demarteau-Ginsburg, E. Lederer, R. Ryhage, S. Stallberg- 
Stenhagen and E. Stenhagen, Nature 183 1117 (1959). 



Aliphatic Acids and Glycolipides 



The metabolic origins of some of the acids in this section can 
be deduced from the foregoing chapter. Among these are 
pyruvic, glyceric, acetic, formic, propionic and lactic acids. 

Many of the other simpler acids are recognizable as members 
of the citric acid cycle and ancillary routes. The citric acid 
cycle (tricarboxylic acid cycle or Krebs cycle) is outlined 
below : 

-' The Citric Acid Cycle 
Enzymes : 

1. Condensing enzyme 

2. Aconitase 

3. Isocitric dehydrogenase 

4. Oxalosuccinic decarboxylase 

5. Succinic dehydrogenase 

6. Fumarase 

7. Malic dehydrogenase 



47 



Aliphatic Acids and Glycolipides 



CH3COCOOH 

Pyruvic Acid 

DPN® 

DPNH^^ — 



^. 



Coenzyme A 
Lipoic Acid, Thiamine Pyrophosphate 



^' 



CO2 



+ H 
CH3CO-C0A 
Acetyl-CoA 
CO— COOH t (T) 

I ' /^ ^r^ I 

CHj— COOH H:0 Co-A HO— C— COOH 



CH2— COOH H.O CH,— COOH 

J I 
< ^^C— ( 



C— COOH 



Oxaloacetic Acid 



DPNH <- 

+ H® 

DPN®^ 



(Z) 



HOCH— COOH 

I 
CH>— COOH 

Malic Acid 



H2O 



® 



HOOC— CH 



Flavin H^ 
Flavin 



HC— COOH 

Fumaric Acid 



® 



CH,— COOH (2) CH— COOH 
Citric Acid c;s-Aconitic Acid 

H2O 




CH2- 


-COOH 


CH— COOH 


HO— CH— COOH 
Isocitric Acid 


^TPN® 

(3) 

J^TPNH + H® 

CH2— COOH 

1 


CH— COOH 

1 


1 

co- 

® 


-COOH 

Oxalosuccinic Acid 


CHo— COOH 


ATP ADP CH2 







CHo— COOH 



CH2— COOH 

Succinic Acid I- \ | 

^xH.O CH2— C00h\c02 i CO— COOH a-Ketoglu- 
CoA-(— ->^| VV,^::^^^ taricAcid 

CH2 /^/^\ Coenzyme A 

CO— CoA I \ 

DPNH DPN® 
Succinyl-CoA -f- 

H© 



Pfizer Handbook of Microbial Metabolites 



48 



The net effect of the cycle is to oxidize pyruvic acid to carbon 
dioxide and water : 

CH3COCOOH + 50 ^ 3CO2 + 2HoO 

Enzymes of the citric acid cycle occur widely among micro- 
organisms, and it is likely that the cycle and variants of it are 
equally ubiquitous. Its primary physiological function in micro- 
organisms (if a primary function can be singled out) is less 
clear, two possibilities being: (a) an energy source and (b) a 
source of amino acid skeletons. Interruption of the cycle or im- 
balances under certain conditions lead to accumulation of cer- 
tain acids. Thus high yields of citric, isocitric, a-ketoglutaric, 
fumaric and malic acids can be obtained in controlled fungal 
fermentations. 

It was mentioned in the preceding chapter that certain micro- 
organisms are capable of growing on a medium containing ace- 
tate as the sole carbon source, synthesizing all their carbo- 
hydrate requirements from it. In some of these microorganisms, 
at least, this ability may be due to possession of a pair of en- 
zymes (malate synthetase and isocitritase ) which permit opera- 
tion of a cycle ancillary to the citric acid cycle or replacement 
of the steps from isocitric acid to malic acid and commonly 
called the glyoxylic acid cycle: 



HC— COOH 

II 

HOOC— CH — 

Fumaric Acid 



The Glyoxylic Acid Cycle 
Acetyl CoA 

CH2— COOH 

■I 

CH2— COOH OHC— COOH 
Succinic Acid Glyoxylic Acid 




HO— CH— COOH 

CH— COOH 

I 

CH2— COOH 
Isocitric Acid 




HO— CH— COOH 
I 

CH2— COOH 
Malic Acid 



CO— COOH 

CH2— COOH 
Oxaloacetic Acid 



CH2— COOH 
I 
HO— C— COOH 

I 

CH2— COOH 
Citric Acid 



49 



Aliphatic Acids and Glycolipides 



The origin of certain other acids can be deduced; for example, 
itaconic acid by decarboxlration of aconitic, oxalic acid by 
oxidation of glyoxyUc and epoxysuccinic by oxidation of fu- 
maric. 



CH— COOH CH2 

II — CO2 II 

C— COOH >C— COOH 



CH2— COOH 
Aconitic Acid 



CHo— COOH 
Itaconic Acid 



OHC— COOH 
Glyoxylic Acid 



[O] 



HOOC H 

\ / 
C=C 

/ \ 

H COOH 

Fumaric Acid 



-* HOOC— COOH 
Oxalic Acid 



[O] 



HOOC 



H 



.U 



H '^' COOH 
frans-Epoxysuccinic Acid 



Certain higher fungi and some molds produce acids such as 
caperatic, agaricic, rangiformic, mineoluteic, roccellic, and 
spiculisporic, which appear to be essentially aldol condensation 
products of various keto acids of the citric acid cycle with long 
chain fatty acids. 



CH3(CH2),3— CH— COOH 

HO— C— COOH 

CH2— COOH 
Caperatic Acid 
(one ccrboxyl group a 
methyl ester) 

CH3(CH2)i3— CH— COOH 
I 
CH— COOH 

I 

CH2— COOH 



Rangiformic Acid 
(one carboxyl group a 
methyl ester) 



CH3{CH2)i5— CH— COOH 

I 
HO— C— COOH 

CH2— COOH 
Agaricic Acid 

CH3(CH2)9— CH C=0 

HOOC— C— OH \ 

I \ 
CH O 

COOH 

Minioluteic Acid 



Pfizer Handbook of Microbial Metabolites 



50 



CHalCHj),!— CH— COOH 

I 

CH— COOH 

CH3 
Roccellic Acid 



CHalCHj),— CH— COOH 

I 
O C— COOH 

/ 

o=c 

\ 

CH2— CH2 

Spiculisporic Acid 



Lipide production by microorganisms varies widely, some 
yeasts and molds producing up to 50% of their dry weight. 
Yeasts were used for commercial submerged culture production 
of fat during World War II in Germany. 

It has been estimated that 80-90% of all fatty acids in plants 
and higher animals occur as esters — triglycerides and phospho- 
lipides. In microorganisms a high percentage of the lipides 
seem to be bound in some way, perhaps as lipoproteins, liposac- 
charides, sterol esters, etc., and a preliminary acid hydrolysis is 
required before complete extraction. 

The fatty acid contents of the fats produced by a few molds 
and yeasts have been studied in detail, and several of these are 
reproduced in the following table. 



TABLE I 
Componenf Faffy Ac/ds of Fofs Produced by Microorganisms 





Asper- 
gillus 
niditlans^ 


Penicil- 

lium 
soppii.'^ 


Penicil- 
lium 

lilaci- 
num^ 


Penicil- 
lium 

spinulo- 
sum* 


Yeast 
Strain 
No. 72^ 


Rhodo- 

torula 

sp.« 


Torulop- 
sis ipj 


Free acidity (% 


08 


0.6 


0.2 


5.8 


33 


18 


51.2 






Component Acids 

Myristic 

Palmitic 


0.7 
20.9 
15.9 

1.4 
1.2 
40.3 
17.0 
0.2 
2.4 


0.3 

22.0 

7.6 

0.9 
3.3 
45.2 
20.0 
0.3 
0.4 


0.1 
32.3 

9.4 

1.4 

3.4 

38.6 

13.4 

1.4 


18.0 
11.9 

1.4 
3.8 
43.3 
21.1 
0.3 
0.2 


0.1 

25.6 

5.9 

5.1 
1.3 
54.5 
5.7 
0.7 
1.1 


1.1 

29.8 

8.8 

1.4 
1.8 
40.1 
11.2 
4.8 
1.0 


0.3 
7.9 
3.8 


Arachidic, Be- 
henic, Ligno- 


0.2 


Hexadecenoic. . . 
Oleic 


7.6 
21.5 




49.7 


Linolenic 

Unsaturated C20. . 


4.4 



51 



Aliphatic Acids and Glycolipides 



Generally microorganism lipides have a higher free fatty 
acid content than those of animals. Bacterial fats seem to have 
received less quantitative study. cis-Vaccenic and lactobacillic 
acids have been shown to be major constituents of the lipides of 
lactobacilli,'* streptococci" and Agrobacterium tiimefaciens}'^ 
An analysis of the fatty acids of two strains of Mycobacterium 
tuberculosis has been published:" 



TABLE II 

Higher Fatfy Acid Content (%) in the Phosphatides and Fats of Mycobacterium tuberculosis 

H.,7 Rv and BCG 





Phosphatide 


Fat 




H:,7Rv 


BCG 


H:,7Rv 


BCG 




20.0 

3.0 

13.8 

5.7 
3.7 

13.0 
28.0 
12.8 


20.4 
3.0 
8.6 

12.3 
14.0 

13.0 
19.2 
10.4 


0.7 

1.1 

2.7 

20.0 

80 

24.5 
34.0 
10.0 






3.1 


II. " " 


2 1 


III. " " 


1.5 


III. Phthioic Acid 


5.5 






IV. " " 






2.3 




22 1 


Oleic and Palmitic Acids 


48.2 




15.2 







The waxes and fats in which the acid-fast mycobacteria and 
corynebacteria abound have been investigated extensively, and 
a variety of oxidized, methylated and branched chain fatty acids 
and alcohols isolated and characterized. In the oxidized and 

ij. Singh, T. K. Walker and M. L. Meara, Biochem. J. 61 85 
(1955). 

2 J. Singh, Sudha E. PhiHp and T. K. Walker, /. Set. Food and Agr. 
8 697 (1957). 

"J. Singh, Sudha Shah and T. K. Walker, Biochem. J. 62 222 
(1956). 

*I. Shimi, Ph.D. Thesis, Univ. of Manchester, 1955. 

^ T. P. Hilditch and R. K. Shrivastava, Biochim. et Biophys. Acta 
2 80 (1948). 

« John Holmberg, Svensk Kem. Tidskr. 60 14 (1948). 

" R. Reichert, Helv. Chim. Acta 28 484 (1945). 

^ Klaus Hofmann and Sylvan M. Sax, /. Biol. Chem. 205 55 
(1953). 

9 Klaus Hofmann and Fred Tauslg, ibid. 213 415 (1955). 

^"Idem., ibid. 213 425 (1955). 

^^ Josef Pokorny, Natiirwissenschaften 10 241 (1958). 



Pfizer Handbook of Microbial Metabolites 52 

methylated acids the oxygen and methyl groups usually appear 
in positions consistent with the acetate theory of fatty acid 
biogenesis. These bacteria seem to be able also (in effect) to 
couple two long chain fatty acids to form ketones and branched 
chain acids. 

Bacterial lipopolysaccharides are irritating pyrogens, relatively 
toxic to higher animals. The polysaccharide component is the 
carrier of serological effects, while the lipide moiety has an 
affinity for the surface of erythrocytes and produces the toxic 
and pyrogenic effect.^- The high molecular weight wax called 
cord factor from mycobacteria is quite toxic (quantitatively 
comparable to diphtheria toxin) and is believed by some to be 
the principal factor responsible for the virulence of tuberculosis 
pathogens. Some of the simpler liposaccharides are shown in 
this section. References to those of higher molecular weight 
are included in an appendix. 

Phosphatides are widely distributed in nature, though gen- 
erally in small quantities. They are difficult to handle intact, 
and few have been well characterized. The metabolism, theories 
of function and biosynthesis of phospholipides have been re- 
viewed. ^^ 

For many years chemists speculated on the reason for the 
predominance of compounds with an even number of carbon 
atoms among natural fatty acids. The mystery was intensified 
by such animal feeding experiments as those of Knoop and 
Dakin," which showed that in mammalian metabolism stepwise 
degradation of fatty acids and similar substances occurred two 
carbon atoms at a time. 

Microorganisms have been instrumental in the discovery of 
the significance of acetate in the cataboUsm and in the biosyn- 
thesis of fatty acids. The enzymatic methods, particularly those 
of anaerobic microorganisms, may differ in detail from those of 
higher animals. This work has been well reviewed. ^^ 

Great advances were made in the discovery of coenzyme A,^*^ 

^- O. Westphal, O. Liideritz, E. Eichenberger and E. Neter, Deut. Z. 
Verdauungs-u. Stoffivechselkrankh. 15 170 (1955). 

13 E. P. Kennedy, Ann. Rev. Biochem. 26 130 (1957). 

1* H. D. Dakin, "Oxidations and Reductions in the Animal Body," 
Longmans, Green and Co., London, 1922. 

'^^ H. A. Barker, "Bacterial Fermentations," John Wiley and Sons, 
Inc., New York, N. Y., 1956, p. 30. 

^® Fritz A. Lipmann, "Les Prix Nobel," Stockholm, 1954. 



53 



Aliphatic Acids and Glycolipides 



the isolation of acetyl coenzyme A (from yeast), the demonstra- 
tion that the acetyl group was attached to its sulfur atom in a 
thioester linkage and that acetyl coenzyme A was an active 
acetylating agent. ^' The enzymic steps in what must be a 
very general scheme of fatty acid catabolism now can be written 
as follows:^® 



MgO 
CoA— SH 



Fatty Acids 
ATP 



AMP + Pyrophosphate 



DPN® 
H© +DPNH 



— CH2— CH2— CH2— CO— S— CoA 
Acyl Coenzyme A 



acy dehydrogenase 



— CH.— CH=CH— CO— S— CoA 
/rans-a,/3-Dehydroacyl Coenzyme A 

enolhydrase 

— CH2— CH— CH2— CO— S— CoA 

1 
OH 



I© 



DPN 



©. 



L-/3-Hydroxyacyl Coenzyme A 



H^ + DPNH 



/3-hydroxyacyldehydrogenase 



— CH,— CO— CH2— CO— S— CoA 
/3-Ketoacyl Coenzyme A 

CoA— SH , 

cleavage enzyme 

— CH2— CO— S— CoA + CH3CO— S— CoA 



At first this process was thought to be reversible or cyclic. It 
has since been shown that a separate set of enzymes controls 
fatty acid biosynthesis. The required enzymes and cofactors 
for the synthetic process have been isolated, and in outline the 

^"^ Feodor Lynen, Ernestine Reichert and Luistraud Rueff , Ann. 574 
1 (1951). 

^* Feodor Lynen, Ann. Rev. Biochem. 24 653 (1955). 



Pfizer Handbook of Microbial Metabolites 



54 



process is at present believed to be represented by the scheme : " 



Biotin 

C02 



CH3CO— S— CoA 
ATP 



ADP 



COOH 



CH2— CO— S— CoA 
Malonyl Coenzyme A 

CH3CO— S— CoA 

COOH 

CH3— CO— CH— CO— S— CoA + HS— CoA 

TPNH (4H®) 



several steps, 
decarboxylase, hydrogenase 



CH3— CH2— CH,— CO— S— CoA + CO, + H,0 

The butyryl coenzyme A can then react with another molecule 
of malonyl coenzyme A and the process repeats. There is a 
statistical distribution peak at 14-18 carbon atom length chains. 
Certain bacteria can couple chains of considerable length as, 
for example, in corynomycolic acid produced by corynebacteria : 



CH3(CH>)u— CH 
OH 



COOH 

I 
CH— (CH,),3CH3 



Corynomycolic 
Acid 



oxidative decarboxylation 



CH3(CH,)u— C— CH,— (CH,),3CH3 + CO. 

O 

This compound is formed by the coupling of two palmitic acid 
molecules as shown by a labeling experiment.-" C^M-Labeled 

13 Salih J. Wakil, Edward B. Titchener and David M. Gibson, 
Biochim. et Biophys. Acta 29 225 (1958); Salih J. Wakil, J. Am. 
Chem. Soc. 80 6465 (1958); David M. Gibson, Edward B. Titchener 
and Salih J. Wakil, Biochim. et Biophys. Acta 30 376 (1958). 

-"Mireille Gastambide-Odier, E. Lederer, Nature 184 1563 (1959). 



55 Aliphatic Acids and Glycolipides 

palmitic acid was incorporated into mycolic acid, and the prod- 
uct degraded to show that the carboxyl group and the oxidized 
C-atom Ij-to it in the corynomycolic acid were labeled. A similar 
biosynthetic path was suggested for the higher molecular weight 
mycolic acids produced by mycobacteria. Thus, condensation 
of 2 moles of n-C-c and 2 moles of n-C,s acids would yield the 
Css mycolic acids of cord factor. A C^f, acid is known to be pro- 
duced by mycobacteria, and a C-.o acid, corynine, by corynebac- 
teria. 

The biotin requirement for enzymatic carboxylations is be- 
coming generally recognized. It was in connection with his 
studies in lipide metabolism that Lynen isolated and synthesized 
a reaction product of biotin and carbon dioxide in which COo 
had reacted at one of the nitrogen atoms to give an allophanic 
acid type of intermediate, the side-chain carboxyl group perhaps 



11 Hooc ;i 

/^\ \ /^\ 

HN N— COOH N NH 

II II 
CH CH or CH CH 

CH2 CH— (CH,),— COOH CH2 CH— (CH2)4— COOH 

being bound to the protein apoenzyme by an amide bond. 

An intermediate may be adenosine diphosphoryl biotin (from 
ATP): 

O OH OH O 

II I I II ^ 

/C\ /P—O—P—O— Adenosine ^Cv /COO© 

HN N ^ ^ HN' N +Adenosine- 

I I + CO2 — > I I diphosphate 

CH CH CH CH 



Other suggestions concerning the detailed function of this 
carboxylase cofactor were made.-' 

The lecithins are formed by initial ATP phosphorylation of 
one glycerol hydroxyl group followed by esterification of the re- 

-^ F. Lynen, J. Knappe, E. Lorch, G. Jutting and E. Ringelmann, 
Angew. Chem. 71 481 (1959). 



Pfizer Handbook of Microbial Metabolites 



56 



maining two hydroxyls by fatty acids as their coenzyme A esters. 
The phosphate group is then displaced by a choline phosphate 
group contributed by a coenzyme, cytidine diphosphocholine : 

O 

II 
CH2— O— C— R 

O CH3 

CH— O— C— R + CH3- 



NHo 



CH2 



O 

T 

-0— P— OH 
OH 



Diglyceride 
Phosphate 



-N— CH2— CH2— O— P- 

I© I 

CH3 eo 



o 

T 
-0— p— o- 

OH 



-CH2 



Cytidine-5'-diphosphatecholme 



/""N 



O 

/On 



OH OH 



CH2— O— C— R 

O 
CH— O— C— R 



+ Cytidine Phosphate 



CH2 



O 

T 



CH3 



-CH2— N— CH3 

©I 

CH3 



-0— P— O— CH2 

00 
The mechanism for cephalin formation is probably similar. 

67 Formic Acid, CHoOo, colorless liquid, b.p. 100.5°, n,,-" 1.3714. 

HCOOH 

Pseiidomonas forniicans n. sp., etc. 

See the reference below for earlier work. 

Irving P. Crawford, /. Bacteriol. 68 734 (1954). 

68 Oxalic Acid, C.H0O4 (Dihydrate), colorless tablets, m.p. 101°. 

HOOC— COOH 

Aspergillus niger, Penicilliuni oxalicinn, Citromyces 
spp., many other fungus species and most lichens. 

It occurs as the calcium salt in most lichens and higher 
fungi, but occasionally also as the free acid. 



57 Aliphatic Acids and Glycolipides 

Jackson W. Foster, "Chemical Activities of Fungi," Aca- 
demic Press Inc., New York, N. Y., 1949, chap. 10, pp. 326- 
350. 

G. Walter, "Organic Acid Production by some Wood-Rotting 
Basidiomycetes," Univ. Microfilms Pub. 10,417, 1955, 99 pp. 

69 Acetic Acid, C0H4O0, colorless liquid, b.p. 118°, nc'" 1.3718. 

CH3COOH 

Saccharomyces cerevisiae, other yeasts. Present in 
small quantities in many microorganisms. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954, Vol. I, Ruse H. Vaughn, Acetic acid-vinegar, chap. 
17, pp. 498-535. 

70 Pyruvic Acid, C;iH403, colorless liquid, b.p. 165° (dec), nn^" 

1.4138. 

CH3COCOOH 

Pseudomonas sac char ophila, etc. 

Approximately 2 moles of pyruvic acid were produced 
per mole of glucose. 

Nathan Entner and Michael DoudorofF, /. Biol. Chem. 196 
853 (1952). 

71 Malonic Acid, C;5H404, colorless plates, m.p. 135°. 

HOOC— CH.— COOH 

PenicilliuTn funiculosum, P. islandicum Sopp, other 
fungi 

D-Mannitol was isolated from the same culture. 
Takeo Yamamoto, J. Pharm. Soc. Japan 75 761 (1955). 

72 Tartronic Acid, C3H4O5, colorless crystals, m.p. 163° (dec). 

HOOC— CH— COOH 

OH 

Acetobacter acetosum, Gluconoacetobacter liquefaciens 
The first organism also produced 2-keto-D-gluconic acid 
and 5-keto-D-gluconic acid. The second organism also 
produced acetaldehyde, formic acid, acetic acid, 5-keto- 
gluconic acid, glycolic acids, other reducing acids, rubigi- 
nol, rubiginic acid and 3,5-dihydroxy-l,4-pyrone. 

D. Kulka, A. N. Hall and T. K. Walker, Nature 167 905 
(1951). 



Pfizer Handbook of Microbial Metabolites 58 

Ko Aida, Toshio Kojima and Toshinobu Asai, /. Gen. and 
Appl. Microbiol. 1 18 (1955). 

73 yg-Nitropropionic Acid, C3H-,04N, colorless crystals, m.p. 65°. 

O2N— CH2CH2COOH 

Aspergillus flavus, A. oryzae 

Milton T. Bush, Oscar Touster and Jean Early Brockman, 
J. Biol. Chevr. 188 685 (1951). 

Seiji Nakamura and Chuji Shimoda, /. Agr. Chem. Soc. 
Japan 28 909 (1954). 

H. Raistrick and A. Stossl, Biochem. J. 68 647 (1958). 

See addendum for reference on biosynthesis. 

74 Propionic Acid, CgHeOo, colorless liquid with sharp odor, b.p. 

140.5°. 

CH3CH2COOH 

Amanita muscaria L., Propionibacteria, Clostridium 
propionicum 

Julius Zellner, Monatsh. 26 727 (1905). 

Kenneth V. Thimann, "The Life of Bacteria," The Macmil- 
lan Company, New York, 1955, pp. 429-440. 

75 L(+)-Lactic Acid (d-Lactic Acid, Sarcolactic Acid), C^H^O.,, 

colorless crystals, m.p. 52.8°, [a],/' +3.33° (c 5.022 in 
water), hygroscopic, polymerizes. 

CH3CHCOOH 

OH 

Lactobacilli, Rhizopus species, etc. 

Yields of 90% or better have been reported. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. I, Ruse H. Vaughn, Acetic acid-vinegar, chap. 
17, pp. 498-535; H. H. Shopmeyer, Lactic acid, chap. 12, 
pp. 391-419. 

76 l(— )-Glyceric Acid, C:^Hf;04, unstable, usually isolated as a salt. 

Ca salt (dihydrate), m.p. 138°, [aW +13.3° (c 4.5 in 
water). 

COOH 

HCOH 

I 
CH2OH 



59 Aliphatic Acids and Glycolipides 

We have observed (by paper chromatographic compari- 
son with an authentic sample on several solvent systems) 
the production of this acid by a wide variety of fungi. It 
is always accompanied by gluconic acid. 

n 2-Phosphoglyceric Acid, C;iH707P. 

COOH 

I 
HC— OPO3H2 

CH2OH 

Yeast 

O. Meyerhof and W. Kiessling, Biochem. Z. 276 239 (1935). 

78 Fumaric Acid, C4H4O4, colorless crystals, m.p. 290° (subl.) 
(dec). 

HOOC H 

\ / 

c=c 

/ \ 

H COOH 

Rhizopus species, also Mucor, Cunnirighamella and Cir- 
cinella species, Aspergillus and Penicillium species, Bole- 
tus spp., Fusaria, etc. 

Yields are about 59 Sr ■ 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. I, Ruse H. Vaughn, Acetic acid-vinegar, chap. 
17, pp. 498-535; Jackson W. Foster, Fumaric acid, chap. 15, 
pp. 470-487. 

79 i-frflrjs-Ethylene Oxide x./S-Dicarboxylic Acid (Epoxy succinic 
Acid), C4H4O-, colorless crystals, m.p. 185° (dec.) [<xW^ 
-117° (c 1 in water). 

HOOC H 



H O COOH 

Aspergillus fumigatus, Monilia formosa, Penicillium 
viniferum 

Yields greater than 20 g. per liter have been obtained. 
Andrew J. Moyer, U. S. Patent 2,674,561 (1950). 



Pfizer Handbook of Microbial Metabolites 60 

80 Succinic Acid, C4H(j04, colorless prisms, m.p. 185-187°. 

HOOC— CH2— CH2— COOH 

Mucor stolonifer, Aspergillus terreus, Ustilina vulgaris, 
Penicillium aurantio-virens, Fusarium oxysporum, lichens, 
etc. 

Occurrence is wide, but yields are generally rather low. 

Ve. S. Butkevich and M. V. Fedorov, Biochem. Z. 219 103 
(1930). 

Jackson W. Foster, "Chemical Activities of Fungi," Aca- 
demic Press Inc., New York, N. Y., 1949, p. 373. 

81 Z-Malic Acid, C4H6O5, colorless crystals, m.p. 99°, [ajn'" -1.43° 

(c 21.65 in water). 

HOOC— CH—CH>— COOH 

OH 

White aspergilli, clasterosporium spp., many other 
fungi. 

Yields are high in some cases. 

Reinhold Schreyer, Biochem. Z. 240 295 (1931). 

John L. Yuill, Chem. Ind. 55 155 (1936). 

82 L(+)-Tartaric Acid, C4H6O6, colorless powder or crystals, m.p. 

168-170° (dec), [aW +11.98° (c 20 in water). 

COOH 

I 
HCOH 

HCOH 

I 
COOH 

Gibberella saubinetii, Acetobacter suboxydans 
Citric and acetic acids were produced also. 
Lyle E. Hessler and Ross A. Gortner, /. Biol. Chem. 119 193 
(1937). 

Jonas Kamlet, U. S. Patent 2,314,831 (1943). 

83 Itaconic Acid, C-^Ht-O^, colorless crystals, m.p. 162-164°. 

CH2=C— COOH 

1 

CH2— COOH 

Aspergillus terreus, Ustilago zeae, Helicobasidium 
monpa, other fungi 



6 1 Aliphatic Acids and Glycolipides 

Jasper H. Kane, Alexander C. Finlay and Philip F. Amann, 
U. S. Patent 2.385.283 (1945). 

Leland A. Undeikofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. I. Lewis B. Lockwood, Itaconic acid, chap. 16, 
pp. 488-498. 

Yields are high in the case of A. terreus. Ustilago zeae 
is reported to produce 15 g. per liter as well as some 
dianthrone and glycolipides. 

R. H. Raskins, J. A. Thorn and B. Boothroyd, Can. J. Micro- 
biol. 1 749 (1955). 

84 fra??s-Ghitaconic Acid, C-,H,;04, colorless needles, m.p. 138°. 

COOH 
CH 

CH 

I 
CHo 

COOH 

Aspergillus niger (on Z-xylose) 

Shinichiro Baba and Kinichiro Sakaguchi, Bull. Agr. Chem. 
Soc. (Japan) 18 93 (1942). 

85 a-Ketoglutaric Acid, C-HfiO.,, colorless crystals, m.p. 115-116°. 

O 

II 
HOOC— C— CH,— CHo— COOH 

Pseudomonas fiuorescens 

Harold J. Koepsell, Frank H. Stodola and Eugene S. Sharpe, 
U. S. Patent 2,724,680 (1955). 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
N. Y., 1954 Vol. II, Lewis B. Lockwood, Ketogenic fermenta- 
tion processes, chap. 1, pp. 18-19. 

86 Dimethylpyruvic Acid, C^H^Og, leaflets, m.p. ~24°, b.p. 76-78°. 

CHs O 

\ II 

CH— C— COOH 

/ 
CHa 



Pfizer Handbook of Microbial Metabolites 62 

Aspergillus spp., Piricularia oryzae ( biotin-deficient 
medium) 

K. Ramachandran and V. Radha, Current Sci. (India) 24 
50 (1955). 

Hirohiko Katsuki, /. Am. Chem. Soc. 77 4686 (1955). 

87 Otlier Keto-Acids: 

Many of the transitory a-keto-acids present in cultures 
of microorganisms can be isolated by means of intercep- 
tors such as 2,4-dinitrophenylhydrazine. One recent pa- 
per reported the following acids identified principally in 
lactic and propionic bacteria cultures: 

Glyoxylic Acid p-Hydroxyphenylpyruvic 
Pyruvic Acid Acid 

a-Ketoisovaleric Acid Hydroxypyruvic Acid 

a-Ketoisocaproic Acid Oxalacetic Acid 

a-Ketocaproic Acid a-Ketoglutaric Acid 

Matti Kreula and Artturi I. Virtanen, Acta. Chem. Scand. 
11 1431 (1957). 

88 Glutaric Acid, C,r,Hs04, colorless needles, m.p. Q?"". 

COOH 

I 
CH2 

CH2 

I 

CH2 

COOH 

Aspergillus niger (on Z-xylose) 

Shinichiro Baba and Kinlchiro Sakaguchi, Bull. Agr. Chem. 
Soc. (Japan) 18 93 (1942). 

89 Ita tartaric Acid, C^HgOfi, occurs as a gummy equilibrium mix- 

ture of lactone and free acid. Characterized as the methyl 
ester derivative. 

COOH 

HO— C— CH,OH 

I 
CHo 

I 
COOH 

Aspergillus terreus mutant 

Frank H. Stodola, M. Friedkin, Andrew J. Moyer and 
Robert D. Coghill, J. Biol. Chem. 161 739 (1945). 



63 Aliphatic Acids and Glycolipides 

90 a-Methylbutyric Acid, C-.HjoOo, colorless crystals, m.p. 176°, 

[a],r^ +17.6°. 

CHaCH-CHCOOH 

CH3 

Peuicillium notatum 

Donald J. Cram and Max Tishler, /. Am. Chem. Soc. 70 
4238 (1948). 

91 a,/i-DihydroxyisovaIeric Acid, C,-,H,o04, colorless syrup, [ccW^ 

-12.4° (c 2 in dilute HCl, pH 1 ) and + 10° (c 2 in water, 
pH 5.5-6.5). Forms crystalline quinine salt. 

CH3 

\ 

C — CH— COOH 

CH3 OH OH 

A valine precursor isolated from a Neurospora crassa 
mutant 

John R. Sjolander, Karl Folkers, Edward A. Adelberg and 
E. L. Tatum, ;. Am. Chem. Soc. 76 1085 (1954). 

92 cis-Aconitic Acid, C,jH,;0,;, colorless crystals, m.p. 125°. 

HC— COOH 

II 

C— COOH 

I 
CH2— COOH 

Aspergillus niger 

This acid presumably is present to some extent in all 
organisms with the citric acid cycle. 

Kinichiro Sakaguchi and Shinichiro Baba, Bull. Agr. Chem. 
Soc. (Japan) 18 95 (1942). (Not isolated) 

93 a«o-Isocitric Acid (Lactone), C,.H„Oe, m.p. 140-141° [aW"" 

+ 42.3° (c 4.83 in water). 

O 

/C. 



COOH CHo 

I " ^ 

J 1/ 

c — c 

H COOH 



Pfizer Handbook of Microbial Metabolites 64 

Penicillium purpiirogeniim Stoll var. rubrisclerotium 
Thorn. 

Yields greater than 20% of the glucose substrate sup- 
plied have been reported. Probably the isomer normal to 
the mammalian citric acid cycle also occurs in some 
microorganisms, but it has not been reported to accumu- 
late. 

Teruhiko Beppu, Shigeo Abe and Kinichiro Sakaguchi, Bull. 
Agr. Chem. Soc. (Japan) 21 263 (1957). 

94 tra ns-/3-Methylglutaconic Acid, C,jHs;04, colorless crystals, m.p. 

131-134°. HOOC— CH=C— CH,— COOH 

CH3 

Ustilago sphaerogena 

This substance is a component of ferrichrome A pig- 
ment,* in which its monohydroxamate is complexed with 
iron. 

Thomas Emery and J. B. Neilands. (In press) 

* See addendum. 

95 Citric Acid, CgHsO- (occurs as monohydrate), colorless crystals 

or white powder, m.p. (monohydrate) ~100°, (anhy- 
drous) 153°. CH,— COOH 

HO— C— COOH 
I 
CH2— COOH 

Wide variety of fungi, e.g., Aspergillus niger. 

Yields are high. 

Leland AT Underkofler and Richard J. Hlckey, "Industrial Fer- 
mentations," Chemical Publishing Co., Inc., New York, N. Y., 
1954 Vol. I; Marvin J. Johnson, The citric acid fermentation, 
chap. 13, pp. 420-445. 

96 Mevalonic Acid Lactone (Hiochic Acid, /^-Hydroxy-yg-methyl- 

S-valerolactone), C,5Hi„0;{, colorless, hygroscopic crystals, 
m.p. 27°, b.p. 90° (0.3 mm.). (Synthetic racemate. ) 

CH3 

CHo I CH,. 
I OH I 
CH, C-O 



65 Aliphatic Acids and Glycolipides 

Yeasts (Isolated from Distillers' Dried Solubles). 

Donald E. Wolf, Carl H. Hoffman. Paul E. Aldrich, Helen R. 
Skeggs, Lemuel D. Wright and Karl Folkers, J. Am. Chem. Soc. 
78 4499 (1956). 

Helen R. Skeggs, Lemuel D. Wright, Emlen L. Cresson, 
Gloria D. E. MacRae, Carl H. Hoffman, Donald E. Wolf and 
Karl Folkers, /. Bad. 72 519 (1956). 

Carl H. Hoffman, Arthur F. Wagner, Andrew N. Wilson, 
Edward Walton, Clifford H. Shunk, Donald E. Wolf, Fred- 
erick W. Holly and Karl Folkers, /. Am. Chem. Soc. 79 2316 
(1957). 

Clifford H. Shunk, Bruce O. Linn, Jesse W. Huff, James L. 
Gilfillan, Helen R. Skeggs and Karl Folkers, ibid. 79 3294 
(1957). 

97 x.^-Dihydroxy/S-methylvaleric Acid, C6H10O4, colorless syrup, 

[a]i,"' +3° (c 2.3 in water containing 1 equiv. of Ca(0H)2) 
and -16.7° (c 2.3 in dilute HCl, pH 1 ). Forms crystalline 
quinine salt. 

CH3 

CH3CH2— C— CH— COOH 

! ! 

OH OH 

A precursor of isoleucine isolated from a Neiirospora 
crassa mutant. 

John R. Sjolander, Karl Folkers, Edward A. Adelberg and 
E. L. Tatum, J. Am. Chem. Soc. 76 1085 (1954). 

98 2-Phospho-4-hydroxy-4-carboxyadipic Acid, C7H11O11P. 

OPO3H2 

HC— COOH 

I 
CHo 

HO— C— COOH 

CH,— COOH 

Escherichia coli 

W. W. Umbreit, J. Bacteriol. 66 74 (1953). 

99 Lipoic Acid (6,8-Thioctic Acid), C8H14O0S2, pale yellow crystals, 

m.p. 47°, [7W +10.4°. 

CH2— CH2— CH— (CH,)4— COOH 

I I 

s s 

Yeast, E. coli mutant 



Pfizer Handbook of Microbial Metabolites 66 

Lester J. Reed, Quentin F. Soper, Geo. H. F. Schnakenberg, 
Stanley F. Kern, Harold Boaz and I. C. Gunsalus, /. Am. Chem. 
Soc. 74 2383 (1952); Lester J. Reed, I. C. Gunsalus, G. H. F. 
Schnakenberg, Quentin F. Soper, Harold E. Boaz, Stanley F. 
Kern and Thomas V. Parke, ibid. 75 1267 (1953). (Isolation) 

Edward Walton, Arthur F. Wagner, Louis H. Peterson, Fred- 
erick W. Holly and Karl Folkers, ibid. 76 4748 (1954); Edward 
Walton, Arthur F. Wagner, Frank W. Bachelor, Louis H. 
Peterson, Frederick W. Holly and Karl Folkers, ibid. 77 5144 
(1955). (Synthesis) 

100 2-Decene-l,10-dioic Acid, CioHigO^, colorless crystals, m.p. 172°, 

HOOC— CH=CH— (CH..)6— COOH 

Penicillium notatum 

Donald J. Cram and Max Tishler, /. Am. Chem.. Soc. 70 
4238 (1948). (Isolation) 

101 10-Undecynoic Acid, CnHigOs, colorless crystals, m.p. 39°. 

HC=C— (CHoJs— COOH 

Rhodotorula glutinis var. lusitanica 

Undecenoic acid was isolated from the same culture. 

Nagueira Prista, Anais. fac. farm. Porto 14 19 (1954). 

102 10-Undecenoic Acid (10-Undecylenic Acid), Ci,Hj„0^., colorless 

crystals, m.p. 24°, n,,'-* 1.4464. 

CH2=CH(CHo)8COOH 

Rhodotorula glutinis var. lusitanica 

Nogueira Prista, Anais. fac. farm. Porto 14 19 (1954). 

103 Myristic Acid, Ci4HoyOj, colorless soft leaflets, m.p. 54°. 

CH3{CH,),oCOOH 
Widely distributed, especially as its triglyceride. 

104 D-^-Hydroxymyristic Acid, C14H2SO.S, colorless crystals, m.p. 73°, 

[a]u'' -16° (c 2.0 in chloroform). 

CH,3{CH>),oCHCH,COOH 

OH 

Escherichia coli 

Obtained together with lauric, myristic and palmitic 
acids from an acid hydrolysate of the phospholipide frac- 
tion. 



67 



Aliphatic Acids and Glycolipides 



Miyoshi Ikawa, J. B. Koepfli, S. G. Mudd and Carl Niemann, 
]. Am. Chem. Soc. 75 1035 (1953). 

105 Mineoluteic Acid, CnjH^c.O;, colorless needles, m.p. 171°, [a]-,4Gi^'' 
+ 108.1° (c 1.07 in acetone) 

-c=o 



CH;,(CH,)9— CH 

HOOC— C— OH 
CH 



COOH 

Penicillium minioluteum Dierckx 
Spiculisporic acid is produced in the same culture. 

John H. Birkinshaw and Harold Raistrick, Biochem. J. 28 
828 (1934). 

106 Palmitoleic Acid (Physetolic Acid, 9-Hexadecenoic Acid), 

CigHsoO^' colorless crystals, m.p. 30-33°. 

CH3(CHo)5CH=CH(CH.)7COOH 

Yeast, Corynebacterium diphtheriae, Streptococcus 
spp., Penicillium lilacinum occurs widely. 

E. Chargaff, Z. phijsiol. Chem. 218 223 (1933). 

Klaus Hofmann and Fred Tausig, /. Biol. Chem. 213 415 
(1955). 

J. Singh, Sudha Shah and T. K. Walker, Biochem. J. 62 222 
(1956). 

107 Pyolipic Acid, CigHaoOy, colorless, viscous oil. 



-CH— CH.— COOH 



HCOH 
HCOH 
HOCH 
CH 



(CH,)6 
CHs 



CHs 

Pseudomonas pyocyanea 
The yield was 1-2 g. per liter. 

Sune Bergstrom, Hugo Theorell and Hans Davide, Arch. 
Biochem. 10 165 (1946). 



Pfizer Handbook of Microbial Metabolites 68 

108 Palmitic Acid, CieHgoOo, soft white crystals, m.p. 62.5°. 

CHslCHoluCOOH 
Widely distributed, especially as esters. 

109 Spiculisporic Acid, CiyHo^O^., colorless crystals, m.p. 145°, [3!]546i 

-14.76° (in alcohol). 

CH3(CHo)a— CH— COOH 

O C— COOH 

/ 

o=c 

\ 

CH2— CH2 

Penicillium spiculisporum Lehman, P. crateriforme 
Gilman and Abbott and P. minioluteum Dierckx 

P. W. Clutterbuck, H. Raistrick and M. L. Pintoul, Trans. 
Roy. Soc. (London) B220 301 (1931). (Isolation and struc- 
ture) 

Albert E. Oxford and Harold Raistrick, Biochem. J. 28 1321 
(1934). (Isolations) 

no Roccellic Acid, C^jH-i.204, colorless crystals, m.p. 131°, [alo^® 
+ 16.80°. 

CH3(CHo)„— CH— COOH 

CH— COOH 

CH3 

Roccella tinctoria (L.), R. viontagnei Bel., etc., also 
Lecanora species 

Yields 1-4%. Erythrin and i-erythritol also were pres- 
ent. 

G. Kennedy, J. Breen, J. Keane and T. J. Nolan, Sci. Proc. 
Roy. Dublin Soc. 21 557 (1937). 

in cis-Vaccenic Acid, Ci,sH;i402, soft white platelets, m.p. 43°. 

CH3(CH,),CH=CH(CH2)9COOH 

Lactobacillus arabinosus, L. casei, Agrobacterium 
tumefaciens. Streptococcus spp. 

Klaus Hofmann, Robert A. Lucas and Sylvan M. Sax, /. Biol. 
Chem. 195 473 (1952). 

Klaus Hofmann and Fred Tausig, ibid. 213 425 (1955). 



69 Aliphatic Acids and Glycolipides 

112 Lactarinic Acid ( 5-Ketostearic Acid), CisH340.{, colorless plates, 

m.p. 87°. 

O 

CH3(CH,),o— C— (CHolsCOOH 

Lactarius rufus Scopol. 

A. K. Schneider and M. A. Spielman, /. Biol. Chem. 142 345 

(1942). 

113 Stearic Acid, CisHj^^Oo, colorless leaflets m.p. 69°. 

CH3(CHo)i6COOH 
Widely distributed. 

114 Lactobacillic Acid (Phytomonic Acid), CigHggOs, colorless crys- 

tals, m.p. 33.6-35°. 

CHslCHola— CH CH— (CHzls— COOH 

\ / 
CH2 

Lactobacillus arabinosus, L. casei, Agrobacterium 
(Phytomonas) tumefaciens 

Klaus Hofmann, Otto Jucker, William R. Miller, Alfred C. 
Young, Jr. and Fred Tausig, J. Am. Chem. Soc. 76 1799 (1954). 

Klaus Hofmann, Gino J. Marco and George A. Jeffrey, ibid. 
80 5717(1958). (Structure) 

115 Tuberculostearic Acid (MO-Methyloctadecanoic Acid), Ci^Hj^sO^, 

colorless oil, m.p. 12.8-13.4°, n,r' 1.4514, [oiW' -0.045°. 

CHslCHo);— CH— (CH-Js— COOH 

I 
CH3 

Mycobacterium tuberculosis var. hominis 

Franklin S. Prout, James Cason and A. W. Ingersoll, /. Am. 

Chem. Soc. 70 298 (1948). (Synthesis) 

116 Alternaric Acid, CoiHgoOs, colorless crystals, m.p. 138°. 

O 
CH3 OH OH CH2 O I 

CH3— CH2— CH— CH— C— CH=CH— CH,— C— CHo— CHo— C— CH CHo 



COOH 



C CH 

O ^ CH3 



Alternaria solani Ell. and Mart., Jones and Grout 



Pfizer Handbook of Microbial Metabolites 70 

John Frederick Grove, /. Chem. Soc, 4059 (1952). (Isola- 
tion) 

J. R. Bartels-Keith and John Frederick Grove, Proc. Chem. 
Soc, 398 (1959). (Structure) 

117 Rangiformic Acid, Cm,H;jsO,;, colorless needles, m.p. 106°, [a]i,-* 

+ 16.2°. 

CH3(CH2),3— CH— COOH 1 

CH — COOH j>Monomethyl ester 

CH2— COOHj 

Cladonia rangiformis Hoffm., C. viitis Sandst. 
Masaru Aoki, J. Pharm. Soc. Japan 66A 52 (1946). 

118 Caperatic Acid, C2iH3g07, colorless leaflets, m.p. 132°, [aW° 

-3.85°. 

CH3(CH2)i3— CH— COOH 

I (one carboxyl group 

HO — C — COOH exists as the methyl 

j ester) 

CHj— COOH 

Parmelia caperata (L.), Nephromopsis stracheyi, f. 
ectocarpisma Hue. 

Protocetraric acid also was present. 

Michizo Asano, Yukio Kameda and Osamu Tamemasa, /. 
Pharm. Soc. Japan 61 203 (1944). 

119 Ungulinic Acid, C^.^H.^sOt;, colorless microcrystalline needles, 

m.p. 78-80°. 

Tentative structure of hydrate (ordinarily a y-lactone) : 

Ri— CH— COOH I Ri=C,6H3.3, R2=R3=H. 

R2— C— COOH II R2=Ci6H33, Ri=R3=H. 

R3— C— COOH Mi R3=Ci6H33, R,=R2=H. 

I 
OH 

Polyporus betulinus 

J. H. Birkinshaw, E. N. Morgan and W. P. K. Findlay, 
Biochem. J. 50 509 (1952). 

Sidonie Marcus, ibid. 50 516 (1952). 



7 1 Aliphatic Acids and Glycolipides 

120 Ajraricic Acid (Agaricin, Laricic Acid, Agaric Acid) C2i.H4„07, 

colorless microcrystalline powder, m.p. 142° (dec), [^tlu^" 
-9° (in dilute NaOH solution). 

CH3(CH.)i5— CH— COOH 

HO— C— COOH 

CH2— COOH 

Polyporus officinalis (=Fomes officinalis, Fomes laricis) 
A yield of 189^ of the weight of the fruiting body has 
been reported. 

H. Thomas and J. Vogelsang, Ann. 357 145 (1907). 

121 Ventosic Acid, C00H44O,., white amorphous powder, m.p. 183°. A 

tetrahydroxybehenic acid. 

Haematomma ventosum, other lichens 
Thamnolic acid was isolated from the same source. 
Yngve Johannes Solberg, Acta Chem. Scand. 11 1477 

(1957). 

122 Tetracosanoic Acid (Lignoceric Acid), C04H4SO0, colorless plates, 

m.p. 87.5°. 

CH3(CH,),,COOH 

Mycobacterium tuberculosis, Phycomyces blakesleeanus, 
Penicillium chrysogenum 

Robert L. Peck and R. J. Anderson, /. Biol. Chem. 140 89 
(1941). 

Karl Bernhard and Hans Albrecht, Helv. Chim. Acta 31 977 
(1948). 

Yoshiro Abe, Proc. Fac. Erig. Keiogijuku Univ. 2 15 (1949). 
(Chem. Abstr. 47 49491) 

123 Pentacosanoic Acid, Co-.H.-.oOs, colorless crystals, m.p. 84°. 

CH3(CHo)o3COOH 

Mycobacterium tuberculosis var. homiriis 
A. Aebi, J. Asselineau and E. Lederer, Bull. soc. chim. biol. 
35 661 (1953). 

124 Hexacosanoic Acid (Phthioic Acid, Cerotic Acid, Cerinic Acid), 

CogHgoOo, colorless crystals, m.p. 88°. 

CH3 (CH..)24COOH 
Mycobacterium tuberculosis, Phycomyces blakesleeanus 



Pfizer Handbook of Microbial Metabolites 72 

Obtained together with palmitic, tuberculostearic and 
mycoceranic acids. 

R. J. Anderson, J. Biol. Chem. 83 505-519 (1929). 

Karl Bernhard and Hans Albrecht, Helv. Chim. Acta 31 977 
(1948). 

Jean Asselineau, Compt. rend. 237 1804 (1953). 

1 25 Mycolipenic Acid ( ( + )-2,4L,6L-Trimethyltetracos-2-enoic Acid ) , 

CotH^-^.Oo, low melting solid [a]i,-" +7.9° (c 25.2 in ether), 
n^"' 1.'4600. 

CH3(CH2)i7— CH— CH,— CH— CH=C— COOH 
CH3 CH3 CH3 

Mycobacterium tuberculosis var. hominis 

J. D. Chanley and N. Polgar, /. Chem. Soc, 1003 (1954). 

(Isolation) 

D. J. Millin and N. Polgar, Proc. Chem. Soc, 122 (1957). 

( Synthesis ) 

126 Csv-Phthienoic Acid (trans-2,4-Dimethyl-13-n-amyl-2-eicosenoic 

Acid), CoyH^oOo, soft white crystals, m.p. 26° and 39° 
( polymorphic ),"[ a] d'' +17.8° ±0.2°, n,,^' 1.4666. 
Tentative structure : 

CHACHoU 

\ 

CH— (CH,)8— CH— CH=C— COOH 

/ I 1 

CH3(CH2)4 CH3 CH3 

The author emphasizes the difference of this compound 

from mycolipenic acid. 

Mycobacterium tuberculosis var. hoviinis 

James Cason, Hans-Ruedi Urscheler and C. Freeman Allen, 

J. Org. Chem. 22 1284 (1957). (Structure) and earher papers 

in this series. 

127 Ustilagic Acids. 

The corn smut fungus produces a group of related 
glycolipides. As originally isolated, the properties of the 
partially purified mixture were given as : CayH^o-eeOiy, color- 
less, needle-like crystals, m.p. 144-147°, [x]rr' +7° (c 1.0 



73 Aliphatic Acids and Glycolipides 

in pyridine). Two of the component structures have been 
characterized as shown: 



H 


OH 




H 


CH2OH 


<^\ 




/ 


\ " 


/ 


OR 


OH 


" U 




Y 


H 




\ 


H 


-o^ 


\o/ 


/\ 


N°" 


H 


4 



CH2OH H OH 

R = — OOC— CH— (CH2)io— CH-CHoOH (Ustilic Acid A) 

OH 

and 
R = — OOC— CH— (CH2)i2— CH— CHoOH (Ustilic Acid B) 

OH OH 

Ustilago zeae, other Ustilaginales spp. 

Yields of 12-33% of the glucose supplied were reported. 

R. H. Haskins and J. A. Thorn, Can. J. Botany 29 585 
(1951). 

R. U. Lemieux, J. A. Thorn, Carol Brice and R. H. Haskins, 
Can. J. Chem. 29 409 (1951). (Isolation) 

R. U. Lemieux, ibid. 29 415 (1951). 

R. U. Lemieux, J. A. Thorn and H. F. Bauer, ibid. 31 1054 
(1953). 

128 Bongkrekic Acid, C29H40OY, unstable, resinous, [ajo" +165° (c 
2.0inNaHCd3). 

The stabler hydrogenated compound, C29H54O7, was 
given the following partial structure. 

C2H5(±CH2) 

HOOCl I 

(C17H33 ± 2CH2)— CH2— C— CH2— CH— CH-COOH 

HOOCj I I , ^ 1 

OCH3 V 

CH3,H 

Pseudomonas cocovenenans (on a special copra-con- 
taining medium) 

Bongkrekic acid is a toxin and has antibiotic properties. 

D. H. Nugteren and W. Berends, Rec. trav. chim. 76 13 
(1957). 



Pfizer Handbook of Microbial Metabolites 



74 



129 Mycoceranic Acid (Mycocerosic Acid), C.^He^O^, white solid, 

m.p. 30°, [a],.-' -6.9° (c 16.8 in ether). " 

CH3(CH2)2iCH— CK2— CH— CHo— CH~COOH 
CH3 CH3 CH3 

Mycobacterium tuberculosis 

Occurs esterified with phthiocerol. 

J. D. Chanley and N. Polgar, /. Chem. Soc, 1003, 1011 

(1954). 

130 Glycolipide from Pseiidomonas aeruginosa, C;{^H,5,|Oi4 (Monohy- 

drate), colorless rectangular platelets, m.p. 86°, [aju —84° 
(c 3.0 in chloroform). 
Probable structure: 



H 

I 
C — 

HCOH 

HCOH O 

I 
HOCH 

CH 



C O— CH— CH,— COO— CH— CH,— COOH 



HCOH (CH.) 

1 i 

— HC CH3 

i 
HOCH 

CH 



(CH,)fi 
CH3 



CH3 



CH3 



Pseudomonas aeruginosa (three different strains) 
F. G. Jarvis and M. J. Johnson, J. Am. Chem. Soc. 71 4124 
(1949). (Isolation) 

131 Corynomycolenic Acid, Co^Hij^O;^, colorless oil, ni/'' 1.4758. 
Methyl ester: [a],-,4(ii"' +9.0 ±0.3°. 

COOH 
CH3(CH,)6CH=CH(CH,)7CHCH(CH,),3CH3 
OH 

Corynebacterium diphtheriae 

J. Pudles and E. Lederer, Biochim. et Biophys. Acta 11 163 

(1953). 



75 Aliphatic Acids and Glycolipides 

132 Corynomycolic Acid, Ch2H,5403, colorless crystals, m.p. 70°, [ajn 

7.5°. 

COOH 

CH3(CH2)i4— CH— CH— (CHo),3CH3 

I 
OH 

Corynebacterhim diphtheriae, C. ovis 

E. Lederer, J. Pudles, S. Barbezat and J. J. Trillat, Bull. soc. 
chim. France 93 (1952). 

Anne Diara and Julia Pudles, Bull. soc. chim. biol. 41 481 
(1959). 

133 Fungal Cerebrins 

A. C42H85O5N 

CHslCHslisCH CH CH CH2OH 

OH OH NH— C— CH(CHo)2iCH3 
O OH 

B. C42H85O6N 

CH3(CH2)i3CH CH CH CHoOH 

OH OH NH— C— CH— CH— (CHskCHs 

II I I 
O OH OH 

PenicilliuTn spp., yeasts 

Takeshi Oda, /. Pharm. Soc. Japan 72 136 (1952). (Isola- 
tion); idem., ibid. 72 142 (1952). (Structure) 

A. H. Cook, "The Chemistry and Biology of Yeasts," A. A. 
Eddy, Aspects of the chemical composition of yeast, Academic 
Press, Inc., New York, N. Y., 1958, p. 203. 

134 Yeast Cerebrin, C44HsoO-,N, colorless crystals, m.p. 87-89°, [a]D 

+31°. 

Tentative structure: 

CH3(CH2),3CH— CH— CH— CH.OH 

! I I 

OH OH NH— C— CH— (CHo)o3CH3 

O OH 
Yeasts 



Pfizer Handbook of Microbial Metabolites 



76 



Fritz Reindel, A. Weichmann, S. Picard, Karl Luber and 
Paul Turula, Ann. 544 116 (1940). 

A. H. Cook, "The Chemistry and Biology of Yeasts," A. A. 
Eddy, Aspects of the chetnical composition of yeast. Academic 
Press, Inc., New York, N. Y., 1958, p. 203. 

135 Lecithins and Cephalins 

The lecithins and cephalins are widely occurring phos- 
pholipides. They are generally oily or partially crystalline 
materials with mixed fatty acids. Lecithin and Cephalin 
Structures (R = various fatty acids). 



a-Lecithin 



CH2— O— R 



CH— O— R 

oe 

CH2— O— P— O— CH2— CH2— N(CH3)3 

II ■ ■ ■ 

O Choline 



© 



/3-Lecithin 
CH2— O— R 

OG 

I 
CH— O— P— O— CH2— CH2— N(CH3)3 

II 
O 

CH2— O— R 



© 



The cephalins are similar except that the choline residue 
is replaced by ethanolamine. 

Yeast, Aspergillus sydoivi, etc. 

F. M. Strong and W. H. Peterson, J. Am. Chem. Soc. 56 952 
(1934). 

D. W. Woolley, F. M. Strong, W. H. Peterson and E. A. Prill, 
ibid. 57 2589 (1935). 

L. F. Salisbury and R. J. Anderson, /. Biol. Chem. 112 541 
(1936). 

136 Dipalmitoleyl-a-lecithin, C40H76O8NP, semi-solid material, [ajn 
+ 6.6°. 

CH2— O— CO— (CHolv— CH=CH— (CH,),— CH3 



CH— O— CO— (CH.JT— CH=CH— (CH,)5— CH3 

O 

II 
CH2— O— P— O— CHo— CHo— N(CH3)3 

I © 

oo 



Yeast 



77 



Aliphatic Acids and Glycolipides 



Donald J. Hanahan and Michael E. Jayko, J. Am. Chem. 
Soc. 74 5070 (1952). (Isolation) 

137 Corynine (Corynodic Acid), C.-2Hn,404, colorless crystals, m.p. 

70°. 

CHa— CH— (CHj);— CH— CH— CH— CH— CH— (CHaliT— CH— (CHJn— CHs 

OH CHs OH CH3 CH.3 COOH CH3 

Corynebacteriiim diphtheriae 

Obtained from the saponification of the phospholipide 
fraction. 

Hideo Takahashi, J. Pharm. Soc. Japan 68 292 (1948). 

138 A Mycolic Acid, CS4H174O4 (^SCH^), colorless microcrystals, 

m.p. 56-58°, [all, +2° (c 2.446 in chloroform). 

OH OH 

CH3— (CH,)„— CH— CH— CH— CH— CH— COOH m + n ~ 28 
(CHoJa R C24H49 R ■ ' C24H49 

CH3 

Mycobactermm tuberculosis human Canetti strain 
This acid was isolated by chromatography from a sa- 
ponification of the chloroform soluble wax. 

Jean Asselineau, Bull. soc. chim. France 135 (1960). 

139 Cord Factor, CisoHaoeOj^ ±10 CH2, nearly colorless wax, m.p. 

43-45°, [a],, +40 ±5° (c 1.37 in chloroform). 

OH 

CH2O— CO— CH— CH— C6oHi2o(OH) 
C24H49 H OH 

H H 




CO— CH— CH— CeuHi-olOH) 

I 
C24H49 



Pfizer Handbook of Microbial Metabolites 78 

Mycobacterium tuberculosis (six different virulent hu- 
man and bovine strains as well as the BCG strain). 

Hydrolysis yields 1 mole of trehalose and 2 moles of 
mycolic acid. 

H. Noll, H. Bloch, J. Asselineau and E. Lederer, Biochim. et 
Biophys. Acta 20 299 (1956). 



Tetronic Acids and Other Lactones 
and Lactams 



This chapter includes derivatives of tetronic acid as well as 
some related lactones. Ascorbic acid is included in this section 
because it is structurally similar to the tetronic acids, although 
it might equally well have been placed with the sugar acids. 

The tetronic acids appear to be condensation products of two 
simple molecules. Ehrensvard and his collaborators have ob- 
tained experimental confirmation of this in two cases.' By 
labeled acetate studies on carlosic and carolic acids, they have 
shown the B portions of the molecules as indicated below to be 



HO 




HO A| B C— CH2— CH2— CHa 



O HOOC— CHo ' O 

Tetronic Acid Carlosic Acid 



B C— CH2— CH2— CH2— OH 




O 
Carolic Acid 

^ Gosta Ehrensvard, "Chemical Society Symposia," Special Publica- 
tion No. 12, The Chemical Society, London, 1958, p. 14. 



Pfizer Handbook of Microbial Metabolites 80 

composed of three acetate units, while the A part is probably 
derived from another source related to carbohydrate biosynthe- 
sis. It would seem as if in the case of carlosic acid the A portion 
were derived from oxaloacetic acid, and in carolic acid from 
lactic or pyruvic acids. 

Inspecting other structures it appears (formally at least) that 
in zymonic acid, isolated by Stodola from many yeasts, the A 
portion could be from tartronate. 

O 

I / il 

HO— C=^=C— CH3 HO— C=^C— C— CH3 

I ! I I i I 

HOOC— CH I C=0 CH \ C=0 

y CH3— CH2— CH z 

I " 

CH3 

Zymonic Acid Tenuazonic Acid 

HOOC— C=4=C— CH3 

I I 

CH3-(CH2)i2— CH ! C=0 

Lichesterinic Acid 
There are other possibilities in this case, however. Tenuazonic 
acid, a lactam similar to the tetronic acids, must surely be de- 
rived from isoleucine and acetoacetate.* Lichesterinic acid 
apparently is the result of a condensation between pyruvate and 
3-oxypalmitate. Nephromopsic acid, which sometimes is found 
with lichesterinic acid, may be a reduction product. 

I 
HOOC— CH CH— CH3 HO— C=[^CH 

CH3— (CH2)i2— CH C=0 CH3— CH i C=-0 

\o/^ ^Y 

Nephromopsic Acid 7-Methyltetronic 

Acid 

OH 

HOOC— CH— C— CH2COOH 

I I 
CH3— (CH2)i2— CH2 C=0 

I 
OH 

Caperatic Acid 

It is interesting to note the co-occurrence of nephromopsic acid 
and caperatic acid, the former being (apparently) a condensa- 
* See addendum. 



8i 



Tetronic Acids and Other Lactones and Lactams 



don product of a C^r, fatty acid and pyruvate while the latter 
seems to be the condensation product of a Ck, fatty acid with 
oxaloacetate. Many other such apparent biosynthetic origins 
can be detected. 

The biosynthesis of penicillic acid has been studied.- At first 
glance this would appear to be derived from acetate and 
dimethylpyruvate, ^-methylglutaconate or a similar unit. It 
was found that 2-C''*-mevalonic acid lactone was not incorporated 
into the penicillic acid molecule when added to the growth me- 
dium of Penicillium cyclophim Westling. However, CH;^C'^OOH 
was incorporated with equal labeling at the sites shown: 



HO 



CH3O— C 
CH3 



=CH 



CH3 



CH2 



X 



C— C 



c=o 



o o 

H 



CHo 



/ 



C— C— C=CH— COOH 



OCH3 



Penicillic Acid 



With a relationship to the terpene biosynthetic route ruled out 
and a similarity to the valine biogenetic pathway also unlikely, 
the authors suggested a precursor of the orsellinic acid type, 
perhaps the 4-methyl ether :t 



CH3 



COOH 



OH 



c 










1 

*CO 

c 


C— CO 


?"' COOH 

1* / 


CO ^ 
CO 

/ 

c 




HO OH 

Orsellinic Acid 






CH3 


- 




CH3 

1 



lo] 


*c 

\/\ * 

C CH— COOH 
*C COOH 

c 


-CO,> 


*C 

C CH2 


CH30 


* *C COOH 

CH3O c 
H 




H 


_J 




Penicillic Acid 



- A. J. Birch, G. E. Blance and Herchel Smith, J. Chem. Soc, 4582 
(1958). 

t See addendum. 



Pfizer Handbook of Microbial Metabolites 



82 



A somewhat similar aromatic ring cleavage has been proposed^ 
in the biosynthesis of patulin. 

It is likely that the biosynthetic origins of the two recently re- 
ported streptomycete antibiotics, acetomycin and 3-carboxy-2,4- 
pentadienal lactol (PA-147) are mutually related. 



CH3 

I 

CH- 

CH 



CH3 

-C— CO— CH3 

I 

c=o 



CH3CO— o o 

Acetomycin 



CH= 

I 
CH 

/ \ / 
HO O 



C— CH=CH2 

c=o 

CH= 



=C— CH=CH2 



O 

3-Carboxy-2,4-pentadienal Lactol 



CH 



c=o 



HO 



The biosynthesis of ascorbic acid in Aspergillus niger is 
known to involve the following stages:* 



000 

II , II II 

:ko c — c — c — 

— — 0= — c 

_ o^ o-^ II c 



COOH CH2OH CH2OH CHoOH 

D-Glucuronic L-Gulono- 2-Keto-L- L-Ascorbic 

Acid lactone gulonolactone Acid 

The glucuronic acid probably quite generally can arise from 
glucose by a hexose interconversion of the type discussed earlier 
in the section on sugars. In muscle tissue it may also come 
from myoinositol. 

•■' J. D. Bu'Lock and A. J. Ryan, Proc. Chem. Soc, 222 (1958). 
*K. Sivarama Sastry and P. S. Sarma, Nature 179 44 (1957). 



83 



Tetronic Acids and Other Lactones and Lactams 



MO y-Methyltetronic Acid, Cr.HfiOg, colorless crystals, m.p. 115°, 



[a].-.«, -21° (c 0.526 in water). 
HO 



CH3 



^o-^X 



Penicillium charlesii G. Smith, P. felliitamnn 

The yield of this and the following tetronic acids from 
P. charlesii totaled l^% of the glucose consumed. 

Percival Walter Clutterbuck, Harold Raistrick and Fritz 
Reutter, Biochem. J. 29 1300 (1935). 

V. C. Vora, /. Set. Ind. Research (India) 13B 504 (1954). 

141 3-Carboxy-2,4-pentadienal Lactol (PA-147), CgHeOg, viscous oil 
which polymerizes on standing at room temperature, [oi]n 
±2° (c2in CHCI3). 



CH=CH2 



CH=CH2 



HC— =C 

C C 

O HO O 




HO O 



Streptomyces sp. 

Hans Els, B. A. Sobin and W. D. Celmer, /. Am. Chem. 
Soc. 80 878 (1958). 

142 Zymonic Acid, CgHeO.-, isolated as the stable methyl ester, b.p. 
118-123° (1 mm.), n,r" 1.4640. 



HO— C= 
HOOC— HC 



<— CH3 

I 

c=o 



Trichosporon capitatum, Hansenula subpelliculosa, 
Kloeckera brevis, Sporobolomyces salmonicolor, Crypto- 
coccus laurentii, Debaryomyces hansenii, Nematospora 
coryli, Torula mellis 



Pfizer Handbook of Microbial Metabolites 84 

Frank H. Stodola, Odette L. Shotwell and Lewis B. Lock- 
wood, /. Am. Chem. Soc. 74 5415 (1952). 

Frank H. Stodola, "Chemical Transformations of Micro- 
organisms," Squibb Lectures on Chemistry of Microbial Prod- 
ucts, John Wiley and Sons, New York, N. Y., 1958, pp. 97- 
102. 

143 Ascorbic Acid (Vitamin C), CgHsOo, colorless crystals, m.p. 
190-192°, [a]ir' +48° (c 1 in methanol). 

HO— C C— OH 

I I 

CH,— CH— CH C=0 

OH OH \^/ 



Serratia marcescens (on xylose), Aspergillus niger 

(Up to 140 mg. per liter yields have been reported from 
A. niger.) 

M. Geiger-Huber and H. Galli, Helv. Chim. Acta 28 248 
(1945). 

Adelheid Galh, Ber. schweiz. botan. Ges. 56 113 (1946). 

J. M. Van Lanen and F. W. Tanner, Jr., Vitamins arid Hor- 
mones 6 163 (1948). 

144 Penicillic Acid, CSH10O4, colorless crystals, m.p. 87° (anhy- 
drous), 64° (hydrate). 



CH3 CH3OV CH3 



C 



CH 






C— C— C=CH— COOH 



HO CH2 OCH3 



Penicillium cyclopium Westling, P. piiberidum Bainier, 
P. thomii, P. baarnense, Aspergillus ochraceus 

John H. Birkinshaw, Albert E. Oxford and Harold Raistrick, 
Biochem. J. 30 394 (1936). (Structure) 

O. F. Black and C. L. Alsberg, 17. S. Dept. Agr., Bur. Plant 
Ind. Bull. No. 199 (1910); Carl L. Alsberg and Otis F. Black, 
Bur. Plant Ind. Bull. No. 270 (1913). (Isolation) 

R. A. Raphael, J. Chem. Sac, 805 (1947). (Synthesis of 
dihydropenicillic acid ) 

E. O. Karow, H. B. Woodruff and J. W. Foster, Arch. 
Biochem. 5 279 (1944). (Isolations) 



85 



Tetronic Acids and Other Lactones and Lactams 



145 Dehydrocarolic Acid, CgHsO^, colorless fine platelets, polymerizes 
above 80°. 



CH. 

I 
O 




CH2 

I 

c 



CH2 o 

Penicillium cinerascens Biourge 

Carlosic acid, spinulosin and gliotoxin also were pro- 
duced. 

A. Bracken and H. Raistrick, Biochem. J. 41 569 (1947). 

146 Carolic Acid, C9Hig04, colorless needles, m.p. 132° [a]-,nn +84° 
(c 0.50 in water). 



CH, 



CH2 

I 
O 



CH, 



\ 



CH3 o 

p. charlesii G. Smith 

Percival W. Clutterbuck, Walter N. Haworth, Harold Rai- 
strick, Geo. Smith and Maurice Stacey, Biochem. J. 28 94 
(1934). 

147 Carolinic Acid, CyHioOg, colorless prisms, m.p. 123° (dec), 
W]:a6i +60° (c 0.34 in water). 



HO 



C— CH2- CH2— COOH 



CH3 o 



Penicillium charlesii G. Smith 



Pfizer Handbook of Microbial Metabolites 86 

L. J. Haynes, J. R. Plimmer, and (in part) A. H. Stanners, 
/. Chem. Soc, 4661 (1956). (Synthesis) 

Percival W. Clutterbuck, Walter N. Haworth, Harold Rai- 
strick, Geo. Smith and Maurice Stacey, Biochem. }. 28 94 

(1934). 

148 Garlic Acid, CioHioO,j, colorless needles, m.p. 176° (dec.) [a:]546i 
-160° (c 0.28 in water). 

CH2 




o=c 

^ CH, 

P. charlesii G. Smith 

Percival W. Clutterbuck, Walter N. Haworth, Harold Rai- 
strick, Geo. Smith and Maurice Stacey, Biochem. J. 28 94 
(1934). (Isolation) 

149 Carlosic Acid, CjoHi^Oc, colorless needles, m.p. 181°, [a].:546] 
-160° (c 0.21 in water). 

O 

HO C— CH2— CH2— CH3 



HOOC— CH2 O 

P. charlesii G. Smith 

Percival W. Clutterbuck, Walter N. Haworth, Harold Rai- 
strick, Geo. Smith and Maurice Stacey, Biochem. J. 28 94 
(1934). (Isolation) 

150 Acetomycin, Ci„Hi40-,, colorless rods, m.p. 115° (subl. 70°), [ajn 
-167° (in ethanol). 

CH3 CH3 

I I 

HC C— CO— CH3 

I I 

CH C 

CH3— CO— o '-' o 

Streptomyces ramulosus n. sp. 



8? 



Tetronic Acids and Other Lactones and Lactams 



The yield was about 1 g. per hter. 

L. Ettlinger, E. Gaumann, R. Hiitter, W. Keller-Schierlein, 
F. Kradolfer, L. Neipp, V. Prelog and H. Zahner, Helv. Chim. 
Acta 41 216 (1958). (Isolation) 

151 Tenuazonic Acid, Ci^Hj-jO^N, straw-colored gum, b.p. 117° 
(0.035 mm.), [a]54Gi'" -136 ±5° (c 0.2 in chloroform). 



HO 



O 

II 

C— CHa 



CH 



CHa— CHj— CH 



\ 



CHa 



Alternaria tenuis Auct. 

Tenuazonic acid is one of several compounds isolated 
from culture filtrates of this fungus. The other substances 
(structures still unknown) were: Altenuic acids I, II and 
III, altenusin, dehydroaltenusin and altertenuol. Altema- 
riol and its methyl ether were isolated from the mycelium. 

T. Rosett, R. H. Sankhala, C. E. Stickings, M. E. U. Taylor 
and R. Thomas, Biochem. J. 67 390 (1957). (Isolation) 

C. E. Stickings, ibid. 72 332 (1959). (Structure) 

152 Terrestric Acid, C11H14O4, colorless crystals, m.p. 89°, [a]546i^° 
+ 61.1° (c 0.53 in water). 



CH2 



CH3— CH,— CH 
O 



CH2 
C 



\ 



CHj O 



Penicillium terrestre Jensen 

John Howard Birkinshaw and Harold Raistrick, Biochem. J. 
30 2194 (1936). 



Pfizer Handbook of Microbial Metabolites 88 

153 Viridicatic Acid (Ethylcarlosic Acid), CisHieOe, colorless plate- 

lets, m.p. 174.5°, [a]n46i'° -105° (c 1.0 in ethanol). 

HO CO— CH2— CH2— CH2— CH,— CHa 

\ / 

C c 

I I 

CH C 

HOOC— CH2 ^ O 

Penicillium viridicatum Westling 

J. H. Birkinshaw and M. S. Samant, Biochem. J. 74 369 
(1960). 

154 Nephrosterinic Acid, C17H28O4, colorless leaflets, m.p. 96°, [<xW° 

+ 10.81°. 

HOOC CH2 

\ / 

CH C 

I I 

CH C 

/ \o/\ 

CH3(CH2)io O 

Nephromopsis endocrocea Asahina (=Cetraria en- 
docrocea (Asahina) Sato) 

Nephrosteranic acid, endocrocin and caperin were also 
present. 

Yasuhiko Asahina, Masaiti Yanagita and Y. Sakurai, Ber. 
70B227 (1937). 

155 Nephrosteranic Acid, C17H30O4, colorless plates, m.p. 95°. 

HOOC CH3 

CH CH 

I 1 

CH C 

CH3(CH2)io ^ O 

Nephromopsis endocrocea Asahina 

Yasuhiko Asahina, Masaiti Yanagita and Y. Sakurai, Ber. 
70B 227 (1937). 

156 Z-Lichesterinic Acid, C19H32O4, colorless needles, m.p. 124°, [cc]v^^ 

-32.66°. 

HOOC CH3 

\ / 

c— c 

I I 

CH C 

CH3(CH2)i2 " O 



Sg Tetronic Acids and Other Lactones and Lactams 

Cetraria islandica f. tenuifolia, Nephromopsis stracheyi 
f. ectocarpisma Hue. 

Yasuhiko Asahina and Masaiti Yasue, Ber. 70B 1053 (1937). 

Yukio Kameda, /. Pharm. Soc. Japan 61 266 (1941). 
(German abstract) 

157 d-Protolichesterinic Acid, C19H32O4, colorless leaflets, m.p. 107.5°, 

[2W +12.1°. 

HOOC CH2 

\ X 

CH C 

I I 

CH C 

CH3(CH2)l2 O 

Cetraria islandica Ach., Parmelia sinodensis Asahina, 
Cladonia papillaria (Ehrh.) Hoffm. 

Yasuhiko Asahina, /. Japan. Botan. 18 489 (1942). 

The Z-isomer, m.p. 107.5°, [a]D'^^ -12.7°, has been iso- 
lated from Cetraria crispa Nyl. (=C. tenuifolia Howe). 

Y. Asahina and M. Asano, J. Pharm. Soc. Japan No. 539, 1 
(1927). 

Eugene E. van Tamelen and Shirley Rosenberg Bach, /. Am. 
Chem. Soc. 80 3079 (1958). (Synthesis) 

158 f-aZio-Protolichesterinic Acid, C19H32O4, colorless plates, m.p. 

107°, [ocW^ -102°. 

Cetraria islandica Ach. var. orientalis Asahina 
Yasuhiko Asahina and Masaiti Yasue, Ber. 70B 1053 (1937). 

159 Nephromopsic Acid, C19H34O4, colorless leaflets, m.p. 137°. 

HOOC CH3 

\ y 

CH CH 

I i 

CH C 

CH3(CHo)i2 '-' O 

Nephromopsis stracheyi f. ectocarpisma Hue. 
Occurs with usnic acid, Michesterinic acid, Z-proto- 
lichesterinic acid and caperatic acid. 

Michizo Asano and Tiaki Azumi, Ber. 68B 995 (1935). 



Carotenes and Carotenoids 



Carotene pigments are widely distributed throughout nature, 
and many microorganism pigments are carotenoid. Their iso- 
lation and characterization are often comphcated by the co-oc- 
currence of closely related compounds, and in some cases by 
poor stability. Many identifications have been made on the 
basis of ultraviolet absorption spectra alone. 

For these reasons, and because of dupHcations in nomencla- 
ture, the literature dealing with microorganism carotenoids is 
confused. The situation has been reviewed by T. W. Goodwin,^ 
and to augment the entries in this section some pertinent tables 
and references from this book have been incorporated as an 
appendix. 

Carotenoids occur in both photosynthetic and non-photosyn- 
thetic microorganisms, and their functions are not established 
clearly. In fungi they may stimulate photokinetic responses 
such as phototropic bending. In sarcina and staphylococcus 
species there may be some protection of the cell from ultraviolet 
light. In photosynthetic genera it has been suggested that 
carotenoids may -serve as blue-light energy absorbers, as oxygen 
carriers and in the prevention of chlorophyll-catalyzed photo- 
oxidations. 

The work that has been done on carotene biogenesis in micro- 
organisms has been well summarized.- ^ It has been found* ^ 

^ T. W. Goodwin, "Comparative Biochemistry of Carotenoids," 
Chemical Publishing Co., Inc., New York, N. Y., 1954. 

- G. E. W. Wolstenholme and Maeve O'Connor, "CIBA Foundation 
Symposium on the Biosynthesis of Terpenes and Sterols," E. C. Grob, 
The biosynthesis of carotenoids by microorganisms, Little, Brown 
and Co., Boston, Mass., 1959, pp. 267-278. 

3 T. W. Goodwin, ibid., pp. 279-294. 



91 



Carotenes and Carotenoids 



that Mucor hiemalis uses acetate for the production of ^-caro- 
tene. The product derived from C"-labeled acetate has been 
partially degraded, and the following partial distribution pattern 
demonstrated : 




i3-carotene 
o Carbon atom from the methyl group of acetate 
• Carbon atom from the carboxyl group of acetate 

Mevalonic acid is an effective carotene precursor in at least 
certain microorganisms. ''■ '' In this connection it is noteworthy 
that in Phyconiyces hlakesleeanus and in Mucor hiemalis the 
production of sterols and carotenoids always runs proportion- 
ally.* The scheme shown below has been proposed for the 
mode of condensation. - 



HOOC- 



OH 



\ 



OH 




COOH 



Leucine has been known for many years to have ketogenic 
and carotenogenic properties to a greater extent than other 
amino acids. The discovery of mevalonic acid facilitated an 



*E. C. Grob and R. Butler, Helv. Chim. Acta 39 1975 (1956). 
■^E. C. Grob, Chimia 10 73 (1956). 

6 G. D. Braithwaite and T. W. Goodwin, Biochem. J. 66 31p (1957). 
^E. C. Grob, Chimia 11 338 (1957). 

* E. C. Grob, M. Bein and W. H. Schopfer, Bull. soc. chim. biol. 
33 1236 (1951). 



Pfizer Handbook of Microbial Metabolites 



92 



explanation of this effect, and this interesting work has been 
reviewed.^' ^° 

Some of the relationships thought to exist are : 



CH, 



CH3 



\ 



o-Ketoglutarate 



CH— CHr- CH— COOH <- 



Glufamate 
CH3 



CoA— SH 



NH2 

Leucine 
CO2 



transaminase, 
pyridoxal CH3 

phosphate 



CH— CH2— C— COOH 



a-Ketoisocaproic Acid 



CH3 



o-ketoacylde- 
hydrogenase, 
thiamin 
pyrophosphate 



CH3 



\ 
< 



CH— CHo- C— S— CoA 



Isovaleryl 
Coenzyme A 



acylde- 
hydrogenase 



flavin 
flavin — H2 



ADP ATP 

O \ biotin f CH3 O 

II v CO2 y \ II 

HOOC— CH2— C=CH— C— S— CoA <-^^^ — ^— ^ C=CH— C— S— CoA 



CH3 

/3-Methylglutaconyl CoA 



H2O 



/3-methyl- / 

glutaconyl CH3 
carboxylase 

/3,/3-Dimethylacrylyl CoA 
(SenecioyI CoA) 



/S-^methyl- 
/^ glutaconase 



CH3 



HOOC— CH2—C—CH2—C—S— CoA 

I 
OH 

^-Hydroxy-/3-methyl 
glutaryl-CoA 

" G. E. W. Wolstenholme and Maeve O'Connor, "CIBA Foundation 
Symposium on the Biosynthesis of Terpenes and Sterols," M. J. 
Coon, F. P. Kupiecki, E. E. Dekker, M. J. Schlesinger and Alice del 
Campillo, The enzymic synthesis of branched-chain acids. Little, 
Brown and Co., Boston, Mass., 1959, pp. 62-74. 

^" Idem., ibid., Harry Rudney, The biosynthesis of P-hydroxy-fi- 
methylglutaryl coenzyme A and its conversion to mevalonic acid, 
pp. 75-94. 



93 



Carotenes and Carotenoids 



Carotenes 





Sterols, Triterpenes, etc. 



cofactors 



CH3 

HOOC— CH2— C— CH2— CH2OH 

I 
OH 

Mevalonic Acid 

± TPNH 
CH3 

HOOC— CH2— C— CH2— CHO 

I 
OH 

Mevaldic Acid 

Y 

± TPNH, DPNH 
CH3 O 

-> HOOC— CH2— C— CH2— C— S— CoA 



condensing 
enzyme 



OH 

/3-Hydroxy-/3-methyl- 

glutaryl-CoA (HMG-CoA) 



HMG 

cleavage 

enzyme 




CH3— C— S— CoA 
Acetyl-CoA 



CH3— C— CHo— C— OH 
Acetoacetic Acid 

succinyl CoA 

transferase 

(ATP activation) 



CH3— C— CH2— C— S— CoA + CoA— SH 
Acetoacetyl CoA 



Fatty Acids 

The precursors of the carotenes are colorless, more reduced 
compounds. These substances then are dehydrogenated in a 
stepwise fashion, a process which requires light and oxygen. 



Pfizer Handbook of Microbial Metabolites 



94 



Oxygen-containing carotenoids appear at an early stage in the 
biosynthetic scheme. Based on the order of appearance in cul- 
tures of Neurospora crassa, Grob has proposed* the following 
partial pathway of carotenoid formation: 




7-Carotene 

Lycopersene has not been isolated from a natural source, but 
this colorless polyene has been synthesized and seems to be a 
logical early member of this sequence. 

* See addendum. 



95 



Carotenes and Carotenoids 



160 Azafrin (Escobedin), C27H;{s04, orange crystals, m.p. 213°, 



h]iu 



-75° (c 0.28 in alcohol), U.V. 428, 458 m^i in 



chloroform. 




COOH 



161 



Mycobacterium phlei 

Mary A. Ingraham and Harry Steenbock, Biochem. J. 29 
2553 (1935). 

Richard Kuhn, Alfred Winterstein and Hubert Roth, Ber. 
64A 333 (1931). 

Torularhodin (May = Lusomycin), C:^7H4s02, red needles, m.p. 
202° (vac.) (dec), U.V. 554, 515, (483) m/A in chloro- 
form. 




COOH 



Rhodotorula rubra, R. sanniei 

The yield from R. sanniei was 2900 y per gram of dry 
cells. Also obtained were torulene (143 y per gram) and 
^-carotene (10 y per gram) and traces of y-carotene and 
lycopene. 

Edgar Lederer, Bull. soc. chim. biol. 20 611 (1938). 

Claude Fromageot and Joue Leon Tchang, Arch. Mikrobiol. 
9 424 (1938). 

L. Nogueira Prista, Congr. Luso-Espan. farm. 2 274 (1952). 
(Chem. Abstr. 48 13807a) 

R. Entschel and P. Karrer, Helv. Chim. Acta 42 466 (1959). 

162 Astacin (3,4,3',4'-Tetraoxo-/3-carotene), C4,jH4s04, violet, metal- 
loid needles, m.p. 240-243°, U.V. 500 m^x in carbon disul- 
fide. 




^^^ 




Mycobacterium laticola 



Pfizer Handbook of Microbial Metabolites 



96 



H. F. Haas and L. D. Bushnell, /. Bacterial. 48 219 (1944). 
(Isolation) 

R. Kuhn, E. Lederer and A. Deutsch, Hoppe-Seylers Z. 220 
229 (1933). 

R. Kuhn and E. Lederer, Ber. 66 448 (1933). 

163 Canthaxanthin (4,4'-Dioxo-;8-carotene) C40H52O2, red crystals, 
m.p. 218°, U.V. 480 nifx, in benzene. 




Cantharellus cinnaharinus, Cory neb acterium michi- 
ganense 

Francis Haxo, Botan. Gaz. 112 228 (1950). (Isolation) 

S. Saperstein and M. P. Starr, Biochem. J. 57 273 (1954). 

F. J. Petracek and L. Zechmeister, Arch. Biochem. and 
Biophys. 61 137 (1956). (Structure) 

C. K. Warren and B. C. L. Weedon, J. Chem. Soc, 3986 
(1958). (Synthesis) 

164 a-Carotene, C40H56, deep purple prisms, m.p. 187.5° (vac), [aW^ 
+ 385° (c 0.08 in benzene), U.V. 446, 473 m^^ in light 
petroleum ether. 




Dacromyces stillatus, Neurospora crassa (mutants), 
Mycobacterium phlei, Phycomyces blakesleeanus, Rho- 
dotorula rubra, Gymno sporangium juniperi-virginianae, 
Puccinia coronifera, Aleuria aurantia, Cantharellus ci- 
barius, Coleosporium senecionis, Penicillium sclerotiorum 

Edgar Lederer, Bull. soc. chim. hiol 20 611 (1938). 

Harry Willstaedt, Svensk. Kern. Tidskr. 49 318 (1937). 

B. L. Smits and W. J. Peterson, Science 96 210 (1942). 

J. Bonner, A. Sandoval, W. Tang and L. Zechmeister, Arch. 
Biochem. 10 113 (1946). 

T. W. Goodwin, Biochem. J. 53 538 (1953). 



97 



Carotenes and Carotenoids 



165 ^-Carotene, C4,|H.r,fi dark violet prisms from benzene-methanol, 
red leaflets from petroleum ether, m.p. 183° (vac.), U.V. 
425, 450, 476 rufi in light petroleum ether. 




^^^^^/'V^^^^^^/'^ 




Phycomyces blakesleeanus, Neurospora crassa, Rho- 
dotorula rubra, R. sanniei, R. glutinis, Sporobolomyces 
roseus, S. salmonicolor, Cantharellus cibarius, C. cinna- 
barinus, Allomyces javanicus, Coleosporium senecionis, 
Mitrula paludosa, Penicillium sclerotiorum, Fremella mes- 
enterica, Puccinia coronifera, Pilobolus bleinii, Gymno- 
sporangium juniperi-virginianae , Dacromyces stillatus, 
Aleuria aurantia, Cryptococcus laurentii, C. luteolus, Mo- 
nilia sitophila, Corynebacterium michiganense (mutants), 
Mycobacterium phlei, Sarcina aurantiaca 

For references see: 

T. W. Goodwin, Ann. Rev. Biochem. 24 497 (1955). 

Idem., "Carotenoids," Chemical Publishing Co., Inc., New 
York, N. Y. 1954, p. 108 etc. 

166 y-Carotene, C40H56, fine deep red crystals with a blue luster from 



benzene-methanol, m.p. 
in petroleum ether. 



177.5°, U.V. 493, 462, 437 nifjL 




Allomyces arbuscula, A. javanicus, A. macrocygna, 
A. moniliformis, Puccinia coronifera, Phycomyces blakes- 
leeanus, Neurospora crassa, Cantharellus cibarius, Coleo- 
sporum senecionis, Dacromyces stillatus, Gymnosporan- 
gium juniperi-virginianae, Cryptococcus laurentii, C. lute- 
olus, Mycobacterium phlei, Chlorobium spp. Penicillium 
sclerotiorum 

For references see: 

T. W. Goodwin, Ann. Rev. Biochem. 24 497 (1955). 



Pfizer Handbook of Microbial Metabolites 98 

Idem., "Carotenoids," Chemical Publishing Co., Inc., New 
York, N. Y. 1954, p. 108 etc. 

J. Bonner, A. Sandoval, W. Tang and L. Zechmeister, Arch. 
Biochem. 10 113 (1946). 

167 8-Carotene, C40H56, fine orange to red needles, m.p. 140.5°, U.V. 
488, 456, 430, 280 m^u in isooctane. 
Proposed structure: 



Cantharellus cibarius, Neurospora crassa (mutants), 
Staphylococcus aureus 

Harry Willstaedt, Svensk Kem. Tidskr. 49 318 (1937). 

Ben Sobin and Grant L. Stahly, /. Bacterial. 44 265 (1942). 

J. W. Porter and M. M. Murphy, Arch. Biochem. and 
Biophys. 32 21 (1951). (Isolation) 

Francis Haxo, Biol. Bull. 103 268 (1952). 

168 Lycopene ( Solanorubin, Rhodopurpurene ) C4oH,-g, brownish red 
to carmine crystals, m.p. 174°, U.V. 446, 474, 506 m^^ in 
petroleum ether. 



Phycomyces Make slee anus, certain Cantharellus spp., 
Neurospora^ crassa. Micrococcus tetragenus (pink type), 
Anthurus aserioformis, Allomyces javanicus, Rhodotorula 
glutinis, R. rubra, R. sanniei, Corynebacterium michiga- 
nense, C. diphtheriae, Mycobacterium phlei. Staphylococ- 
cus aureus, Coleosporium senecionis, Sarcina aurantiaca 

Harry Willstaedt, Svensk. Kem. Tidskr. 49 318 (1937). 

Francis Haxo, Arch. Biochem. 20 400 (1949). 

P. Karrer, C. H. Eugster and E. Tobler, Helv. Chim. Acta 
33 1349 (1950). (Synthesis) 

T. W. Goodwin, Ann. Rev. Biochem. 24 497 (1955). 

Synnove Liaaen Jensen, Germaine Cohen-Bazire, T. O. M. 
Nakayama and R. Y. Stanier, Biochim. et Biophys. Acta 29 
477 (1958). 



99 



Carotenes and Carotenoids 



169 



Rhodopin, C4„H,:^60, violet-red needles, m.p. 168° (171°), U.V. 
440, 470, 501 m^ in light petroleum. 




Polystigma rubrum 

Edgar Lederer, Bull. soc. chim. biol. 20 611 (1938). 

Synnove Liaaen Jensen, Acta Chem. Scand. 13 842 (1959). 
(Structure) 

Paul Karrer and Ulrich Solmssen, Helv. Chim. Acta 18 25, 
1306 (1935); 21 454 (1938). 

170 Rubixanthin (3-Hydroxy-y-carotene), C40H56O, coppery red nee- 
dles, m.p. 160°, U.V. 432, 462, 494 nifj, in hexane. 



^^^ 





171 



Staphylococcus aureus, Coleosporium senecionis. Mi- 
crococcus tetragenus 

E. Lederer, Bull. soc. chim. biol. 20 611 (1938). 

Ben Sobin and Grant L. Stahly, J. Bacteriol. 44 265 (1942). 

H. A. Reimann and C. M. Eklund, J. Bact. 42 605 (1941). 

Richard Kuhn and Chrlstoph Grundmann, Ber. 67 339 
(1934). 

Cryptoxanthin (Cryptoxanthol, 3- or 4-Oxy-/?-carotene, C40H56O) 
deep red prisms, m.p. 169° (vac), optically inactive, U.V. 
425s, 450, 480 m/x in hexane. 





^^^ 



Mycobacterium phlei, Dacromyces stillatus, Vibrio 
adaptatus, Pseudomonas xanthochrus, P. aestumarina, 
Rocella montagnei 



Pfizer Handbook of Microbial Metabolites lOO 

Richard Kuhn and Christoph Grundmann, Ber. 66 174 
(1933). 

Mary A. Ingraham and Harry Steenbock, Biochem. J. 29 
2553 (1935). 

F. P. Zscheile, J. W. White, B. W. Beadle and J. R. Roach, 
Plant Physiol. 17 331 (1942). 

T. R. Seshadry and S. S. Subramanian, Proc. Indian Acad. 
Sci. 30A (1949). 

172 Lycophyll (3,3'-Dihydroxylycopene), C40H56O2, purple crystals, 
m.p. 179°, U.V. 444, 473, 504 m^ in petroleum ether. 




HO— 




Rho do spirillum ruhrum, Chromatium spp. 
M. S. Barber, L. M. Jackson and B. C. L. Weedon, Proc. 
Chem. Soc, 96 (1959). (Structure) 

L. Zechmeister and L. V. Cholnoky, Ber. 69B 422 (1936). 

173 Zeaxanthin (Zeaxanthol), C40H56O2, yellow crystals, m.p. 207° 
(215°), optically inactive, U.V. 451, 476 m/x in petroleum 
ether. 

/OH 

A 



M' ' ' ' 

HO^ ' 

Mycobacterium phlei, Dacromyces stillatus, Staphylo- 
coccus aureus, Pseudomonas xanthochrus, P. aestumar- 
ina. Vibrio adaptatus 

Erwln ChargafF and Joseph Dieryck, Naturwissenschaften 
20 872 (1932). 

Mary A. Ingraham and Harry Steenbock, Biochem. J. 29 
2553 (1935). 

Walter Steuer, Zentr. Bakteriol. Parasitenk. 167 210 (1956). 

T. W. Goodwin, Biochem. J. 53 538 (1953). 



lOI 



Carotenes and Carotenoids 



174 Lutein (Xanthophyll, Luteol), C40H56O2, yellow prisms, m.p. 
190°, [alcd'" +165° (c 0.7 in benzene), U.V. 420, 446.5, 
476 m^u, in petroleum ether. 




Mycobacterium phlei, Staphylococcus aureus, Sarcina 
hitea, Micrococcus lysodeikticus 

Erwin Chargaff, Compt. rend. 197 946 (1933). 

Mary A. Ingraham and Harry Steenbock, Biochem. J. 29 
2553 (1935). 

Tatsuo Ohta, J. Pharm. Soc. Japan 71 1319 (1951). (Isola- 
tion) 

A. R. Gilby and A. V. Few, Nature 182 55 (1958). 

175 Neurosporene (6,7,6',7'-Tetrahydrolycopene), C40H60, yellow-or- 
ange or yellow-brown crystals, m.p. 124°, U.V. 414, 438.5, 
469 rrifj. in petroleum ether. 




Neurospora crassa, Rhodotorula rubra, etc. 

Neurosporene and hydroxylated neurosporenes are 
probable intermediates in the biogenesis of other carote- 
noids occurring in microorganisms. 

J. Bonner, A. Sandoval, W. Tang and L. Zechmeister, Arch. 
Biochem. 10 113 (1946). 

Francis Haxo, ibid. 20 400 (1949). 

L. Zechmeister and B. Kenneth Koe, /. Am. Chem. Soc. 76 
2923 (1954). 

Synnove Liaaen Jensen, Germaine Cohen-Bazire, T. O. M. 
Nakayama and R. Y. Stanler, Biochim. et Biophys. Acta 29 477 
(1958). 



Pfizer Handbook of Microbial Metabolites 



I02 



176 7;-Carotene, C4,,H,j4, probably has not been entirely purified, U.V. 
376 (380), 396 (404), 418 (424), 450. 




Phycomyces blakesleeanus, Neurospora crassa (mu- 
tants), Dacromyces stillatus 

H. A. Nash and F. P. Zscheile, Arch. Biochem. 7 305 
(1945). 

T. W. Goodwin, "Carotenoids," Chemical Publishing Co., 
Inc., New York, N. Y. 1954, p. 108, etc. 

G. MacKinney, C. O. Chichester and Patricia S. Wong, 
Arch. Biochem. and Biophys. 53 480 (1954). 

177 Phytoene (7,8,ll,12,12',ll',8',7'-Octahydrolycopene), C40H64, 
colorless, viscous oil with a strong fluorescence in ultra- 
violet light, U.V. 275s, 283, 295s in isooctane. 




^^^/-^ 



Mycobacterium phlei, Rhodopseudomonas spheroides 
(mutant), Rho do spirillum rubrum 

J. W. Porter and F. P. Zscheile, Arch. Biochem. and Biophys. 
10 537 (1946). 

W. J. Rabourn and F. W. Quackenbush, Arch. Biochem. and 
Biophijs. 61 111 (1956). (Structure) 

T. W. Goodwin, and Malini Jamikorn, Biochem. J. 62 269 
(1956). 

178 Phytofluene (5,6,7,8,9,10,10',9',8',7',6',5'-Dodecahydrolycopene), 
C4„Hfis, colorless, viscous oil with a strong fluorescence in 
ultraviolet light, U.V. 332, 347, 367 m^u, in petroleum 
ether. 




Neurospora crassa, N. sitophila, Mycobacterium phlei, 
Phycomyces blakesleeanus, etc. 



I03 



Carotenes and Carotenoids 



Phytofluene probably occurs widely among microorgan- 
isms. It is a probable precursor of many of the carotene 
pigments. 

L. Zechmeister and F. Haxo, Arch. Biochem. 11 539 (1946). 
(Isolation from neurospora) 

L. Zechmeister, Experientia 10 1 (1954). (Structure) 

179 P-481,C4iH-,sO, U.V. 455, 482, 514 m^ in petroleum ether. 
Tentative structure: 




V^^^%^^/^^/^==^ /^ 



OCH3 



Rho do spirillum rubriim, Chromatium spp. 
M. S. Barber, L. M. Jackson and B. C. L. Weedon, Proc. 
Chem. Soc, 96 (1959). (Structure) 

Synnove Liaaen Jensen, Acta Chem. Scand. 12 1698 (1958). 

180 Hydroxy-P-481 (May = Rhodovibrin), C41H5SO., U.V. 455, 482, 
515 m/x in petroleum ether. 
Tentative structure: 




^^-"^^-^^ 



OH 



Rho do spirillum rubrum, Chromatium, spp. 
M. S. Barber, L. M. Jackson and B. C. L. Weedon, Proc, 
Chem. Soc, 96 (1959). 

Synnove Liaaen Jensen, Acta Chem. Scand. 12 1698 (1958). 

181 Hydroxyspirilloxanthin (May = Bacteriopurpurin, Bacterioeryth- 
rin) C4iH-„sOo, U.V. 489, 523 m^x in petroleum ether. 
Tentative structure: 




^^^^.^5^/^^^ 



HO 

\l 



OCH3 

Rhodospirillum rubrum, Chromatium spp. 



Pfizer Handbook of Microbial Metabolites 1 04 

M. S. Barber, L. M. Jackson and B. C. L. Weedon, Proc. 
Chem. Soc, 96 (1959). 

182 Pigment R ( Spheroidenone ) , C41H58O2, red crystals, m.p. 155.5- 
158°, U.V. 460 (455), 482 (4"87), 513 (516.5) rrifi in 
light petroleum. 
CH3O 

I I 





Rhodopseudomonas spheroides, other purple bacteria 
C. B. Van Niel, Antonie Van Leeuwenhoek J. Microbiol. 
Serol Jubilee Vol. Albert J. Kluyver 12 156 (1947). (Isola- 
tion) 

T. W. Goodwin, D. G. Land and M. E. Sissins, Biochem. J. 
64 486 ( 1 956 ) . ( Structure ) 

183 Pigment Y, C41H60O, yellow unstable crystals, m.p. 116-135° 
(dec). Stable in solution. U.V. 426.5, 452 (454), 484 
(486) nifj. in petroleum ether. 
CH3O I 




^^^/^/^==^^^^^^^ 



Rhodopseudomonas spheroides, other purple bacteria 

A hydroxylated pigment Y was produced in the same 
fermentation, but could not be crystallized. 

C. B. Van Niel, Antonie Van Leeuwenhoek J. Microbiol. 
Serol. Jubilee Vol. Albert J. Kluyver 12 156 (1947). (Isola- 
tion ) 

T. W. Goodwin, D. G. Land and M. E. Sissins, Biochem. J. 
64 486(1956). (Structure) 

Synnove Liaaen Jensen, Acta Chem. Scand. 12 1698 (1958). 

184 Spirilloxanthin (Rhodoviolascin), C42H6„02, violet spindle-form 
crystals, m.p. 218°, U.V. 464, 491, 524 m^x in petroleum 
ether. 

CH3O 

\l 




^/^=^-/W^'^/^^^/^^/^^/^-^ 



1 05 Carotenes and Carotenoids 

Rhodospirillum rubrum, other purple bacteria, Neuro- 
spora crassa (mutants), Chromatium spp. 

P. Karrer and U. Solmssen, Helv. Chim. Acta 18 1306 
(1935). 

C. B. Van Niel and James H. C. Smith, Arch. Mikrobiol. 6 
219 (1935). (Isolation) 

A. Polgar, C. B. Van Niel and L. Zeehmeister, Arch. 
Biochem. 5 243 (1944). 

Synnove Liaaen Jensen, Germaine Cohen-Bazire, T. O. M. 
Nakayama and R. Y. Stanier, Biochim. et Biophys. Acta 29 
477 (1958). (Synthesis) 

M. S. Barber, L. M. Jackson and B. C. L. Weedon, Proc. 
Chem. Soc, 96 (1959). 

185 Torulene, C42H60O2, dark red crystals, m.p. 185°, U.V. 460, 486, 
519 m^ in petroleum ether. 

Tentative structure: 

OCH3 



CH30 



Rhodotorula rubra 

Occurs together with y8-carotene, torularhodin and an 
unstable, uncharacterized carotene. 

Edgar Lederer, Bull. soc. chim. biol. 20 611 (1938). 

J. Bonner, A. Sandoval, W. Tang and L. Zeehmeister, Arch. 
Biochem. 10 113 (1946). 

186 Sarcinaxanthin, yellow crystals, m.p. 149°, U.V. 415, 440, 469 
m/x in petroleum ether. 

About 3.4 mg. of this mono-hydroxy xanthophyll were 
obtained from 385 g. of dried Sarcina lutea cells. It is 
also produced by Flavobacterium marinotypicum and by 
Staphylococcus citreus. 

A closely related hydrocarbon, sarcinene, occurs in all 
these species as well as in Flavobacterium sulfureum. 

Yoshiharu Takeda and Tatuo Ota, Z. physiol. Chem. 268 1 
(1941). (Isolation) 

Doris P. Courington and T. W. Goodwin, J. Bacterial. 70 
568 (1955). 

Tatsuo Ohta, Toshio Miyazaki and Teruo Minomiya, Chem. 
Pharm. Bull. 7 254 (1959). 





Pfizer Handbook of Microbial Metabolites io6 

187 Neurosporaxanthin, dark grayish purple leaflets, m.p. 192° 

(vac), U.V. 472 m^ in hexane (486 m^ in benzene). 

An uncharacterized carotenoid which gives yellow so- 
lutions and a red color adsorbed on sucrose. 

Neiirospora crassa 

Marko Zalokar, Arch. Biochem. and Biophys. 70 568 
(1957). (Isolation) 

188 Leprotene (Leprotin), coppery red needles, m.p. 197°, U.V. 429, 

452, 479 rriyu in petroleum ether. 

The principal carotene of Mycobacterium phlei and 
other mycobacteria. It contains no ionone rings and does 
not function as a provitamin A. 

Yoshiharu Takeda and Tatsuo Ohta, J. Biochem. Japan 36 
535 (1944). (Isolation) 

Tatsuo Ohta, /. Pharm. Soc. Japan 71 462 (1951). 

189 Mycoxanthin, U.V. 385, 406, 430 m^ in petroleum ether. 

A new yellow carotenoid with a relatively short chromo- 

phore. 

Mycobacterium phlei, M. marianurn, M. battaglini 
Aldo Gaudiano, Atti. accad. nazl. Lincei, Rend., Classe set. 

fis., mat. e nat. 21 308 (1956). (Chem. Abstr. 51 8876 f) 

(Isolation) 



Polyenes and Polyynes, 

Excluding Polyene Macrolides 



The polyenes of this section somewhat resemble crocetin, 
bixin and the carotenes in their long systems of conjugated 
double bonds with the resultant color and other physical prop- 
erties, but they lack the isoprenoid structure. 

The acetylenic compounds often occur in low yields and in 
complex mixtures. While generally colorless, they are conspic- 
uous by their strong and characteristic ultraviolet absorption 
spectra. Many of them are unstable. 

From the examples reported to date it seems that basidiomy- 
cetes are the principal producers of such metabolites among 
microorganisms, although such substances occur widely in 
higher plants. That lower fungi are capable of forming poly- 
enes is demonstrated, however, by the side-chains of metabolites 
classified elsewhere, for example fumagillin, sorbicillin and 
auroglaucin : 



C— (CH=CH)o— CHs 




C— (CH=CH)4— COOH 

II 
O 



Fumagillin Sorbicillin 



Pfizer Handbook of Microbial Metabolites 1 08 



CH3(CH=CH) 




OH 

Auroglaucin 



It is likely that both polyenes and polyynes are acetate-de- 
rived. It has been demonstrated^ that nemotinic acid with 11 
carbon atoms is formed from 6 moles of an acetic acid deriva- 
tive, with head to tail Hnkage and elimination of the terminal 
methyl group. 

HC^C— C=C— CH=C=CH— CH— CHj— CH2— COOH 

OH 

Nemotinic Acid 

CH3— C=C— C=C— CH=C=CH— CH— CH2— CHo— COOH 

OH 
Odyssic Acid 

Odyssic acid was presumed to be formed similarly, but with 
terminal methyl group retention. 

In the examples available the acetylenic acids with an odd 
number of carbon atoms terminate in an acetylenic bond. This 
seems to indicate elimination of the terminal methyl group by 
oxidation and decarboxylation. It is interesting to note that the 
reverse process has been demonstrated in the conversion of 
propynoic acid to acetylenedicarboxylic acid by a soil isolate.^ 

CO2 + HC^C— COOH -^ HOOC— C=C— COOH 

The xyloside of nemotinic acid also has been isolated.^ When 
isolated from a culture grown on glucose with l-C"-labeled 
acetic acid added to the medium, labeling is found in the poly- 
acetylenes but not in the xylose moiety. When isolated from 

1 J. D. Bu'Lock and H. Gregory, Biochem. J. 72 322 (1959). 

2 Akira Hanaoka, Tokuya Harada and Takeo Takizawa, /. Agr. 
Chem. Soc. Japan 26 151 (1952). 

3 J. D. Bu'Lock and H. Gregory, Experientia 15 420 (1959). 



log Polyenes and Polyynes, Excluding Polyene Macrolides 
HC=C— C^C— CH=C=CH— CH— CH2— CH2— COOH 




a culture grown on ethanol with l-C^*-labeled acetic acid added 
to the medium, labeling was found in the xylose as well as in 
the acetylenic acid. It was assumed that in the latter case, 
where the molecule was synthesized entirely from C2 units, the 
xylose was produced by way of intermediates closely related to 
glucose. Glucose itself acted as the xylose precursor, then, in 
the first experiment. A closer analysis of the labeling pattern 
of the xylose moiety led to the suggestion that the pentose was 
formed from glucose by way of glucuronic acid followed by 
decarboxylation. 

Many of the acetylenic acids have antibiotic properties. 
A review of polyacetylenes was published recently.* 
About a dozen more compounds of this type are listed in the 
addendum. 

190 Agrocybin, C8H5O0N, unstable compound white crystals, darken- 

ing in air, m.p. 130-140° (dec. explosively), U.V. 216, 
224, 269, 286, 304, 325 m^u in 95% ethanol. 

HOCH.— C=C— C^C— C=C— CONH2 

Agrocyhe dura 

Marjorie Anchel, J. Am. Chem. Soc. 74 1588 (1952). 

J. D. Bu'Lock, E. R. H. Jones, G. H. Mansfield, J. W. Thomp- 
son and M. C. Whiting, Chem. and Ind., 990 (1954). (Struc- 
ture) 

P. J. Ashworth, E. R. H. Jones, G. H. Mansfield, K. Schlogl, 
J. M. Thompson, M. C. Whiting, /. Chem. Soc, 950 (1958). 
(Synthesis) 

191 Diatretyne 1, CsHjOgN, unstable crystals, m.p. 198° (dec. ex- 

plosively), U.V. 223, 260, 275, 290, 309 m^ m 95% etha- 
nol. 

HOOC— CH=CH— C^C— C=C— CONH2 
and 

4E. R. H. Jones, Proc. Chem. Soc, 199-211 (1960). 



Pfizer Handbook of Microbial Metabolites no 

192 Diatretyne 2 (Nudic Acid B), C^HgOoN, short colorless needles, 

m.p. 179^ (dec), U.V. 228, 238, 268, 283, 299, 322 iti/a 
in 957c ethanol. 

HOOC— CH=CH— C=C— C=C— C=N 

Clitocybe diatreta 

Marjorie Anchel, J. Am. Chem. Soc. 74 1588 (1952). 

Idem., ibid. 75 4621 (1953). 

Idem. Science 121 607 (1955). (Structure) 

P. J. Ashworth, E. R. H. Jones, G. H. Mansfield, K. Schlogl, 
J. M. Thompson and M. C. Whiting, J. Chem. Soc, 950 (1958). 
(Synthesis) 

193 trans-Non-2-ene-4,6,8-triyn-l-al, C9H4O, colorless needles, which 

rapidly decompose in light at room temperature. U.V. 
210.5 (220), 228, 240, 257, 271, 287, 306, 327 m/j, in 
ethanol. 

HC=C— C=C— C=C— CH=CH— CHO 

Coprinus quadrifidus 

Six related compounds occurred in the same culture. 
E. R. H. Jones and J. S. Stephenson, /. Chem. Soc, 2197 
(1959). 

194 trans-Non-2-ene-4,6,8-triyn-l-ol, CgH^O, colorless crystals, decom- 

posing at ordinary conditions, U.V. 233, 243, 255, 283, 
300, 320 m,x in hexane. 

HC=C— C^C— C=C— CH=CH— CHoOH 

Coprinus quadrifidus 

E. R. H. Jones and J. S. Stephenson, /. Chem. Soc, 2197 
(1959). 

195 (2d,3d)-Nona-4,6,8-triyn-l,2,3-triol, CjHsO.j, colorless crystals 

(dec.) ~40°, [a],. +6° (c 0.82 in ethanol), U.V. 208, 254, 
269.5, 286.5, 305 m^^ in ethanol. 

HC=C— C=C— C^C— CHCHCH:OH 

I I 
OH OH 

Coprinus quadrifidus 

E. R. H. Jones and J. S. Stephenson, J. Chem. Soc, 2197 
(1959). 



Ill Polyenes and Polyynes, Excluding Polyene Macrolides 

196 Biformin, highly unstable crystals. 

Probably a straight-chain, nine carbon atom glycol, con- 
taining two acetylenic and two ethylenic bonds in con- 
jugation. 

Polyporus bifonnis 

A similar substance, biforminic acid, occurred in the 
same culture. 

William J. Robblns, Frederick Kavanagh and Annette 
Hervey, Proc. Nat. Acad. Sci. 33 176 (1947). 

Marjorie Anchel and Marvin P. Cohen, /. Biol. Chem. 208 
319 (1954). 

197 tra?2S-Dec-2-ene-4,6,8-triyn-l-al, CjoHuO, pale yellow needles, m.p. 

108°, U.V. (225) (234.5), 245.5, 258 (272), 288, 306, 
326, 350, m^ in hexane. 

CH3— C^C— C=C— C=C— CH=CH— CHO 

Pleurotus ulmarius 

J. N. Gardner, E. R. H. Jones, P. R. Leeming and J. S. 
Stephenson, J. Chem. Soc, 691 (1960). 

198 Diatretyne-3 (traris-10-Hydroxydec-2-ene-4,6,8-triynoic Acid), 

Ci„H«;03^ nearly colorless rods from ethyl acetate, rapidly 
becoming coated with blue-green polymer, U.V. 253, 280, 
297, 316, 339 m^. 

HOCH2— C=C— C^C— C^C— CH=CH— COOH 

Clitocybe diatreta 

The author noted the similarity to the antibiotic prin- 
ciple of the royal jelly of bees: 

trans 
HOCH,CH2CH2CH,CHoCHoCH2— CH=CH— COOH 

Helen Flon and Marjorie Anchel, Arch. Biochem. and 
Biophys. 78 111 (1958). 

Marjorie Anchel, Arch. Biochem. and Biophys. 85 569 
(1959). 

199 Deca-trarzs-2,tra?is-8-diene-4,6-diyne-l,10-dioic Acid, Ci„Hg04, 

amorphous powder, m.p. (dec.) ^200°, U.V. 216 (258), 
267, 296, 315, 338 m^^ in ethanol. 

HOOC— CH=CH— C^C— C=C— CH=CH— COOH 

Polyporus anthracophilus 



Pfizer Handbook of Microbial Metabolites 112 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, J. Chem. 
Soc, 1607 (1957). 

200 Marasin [(-)-Nona-3,4-diene-6,8-diyne-l-ol], CioHgO, unstable 

oily substance, polymerized spontaneously, [<x]d^^ about 
-325° (c 0.2). 

HC^C— C=C— CH=C=CH— CH2— CH2— OH 

Marasmius ramealis 

Gerd Benz, Arkiv for Kemi 14 305 (1959). 

201 trans-Dec-2-ene-4,6,8-triyn-l,10-diol, CjoHsO^, colorless needles, 

(dec.) 138°, U.V. 205, 212, 231, 243.5, 259, 279, 290.5, 
309.5, 330.5 m^ in ethanol. 

HOCH2— C^C— C=C— C=C— CH=CH— CH2OH 

Coprinus quadrifidus 

E. R. H. Jones and J. S. Stephenson, /. Chem. Soc, 2197 
(1959). 

202 trans,trans-Matricaria Acid, CioHgOa, colorless plates, m.p. 175° 

(dec), U.V. 245, 256, 310, 3"29 rri/x in ethanol. 

CH3— CH=CH— C=C— C=C— CH=CH— COOH 

Polyporiis anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, J. Chem. 
Soc, 1607 (1957). 

203 trans, trans-Matricarianol, CiyHi„0, colorless needles, m.p. 

105.5°, U.V. 217.5, 231.5, 237, 247, 261, 276, 293, 312 
m/x in ethanol. 

CH3— jCH=CH— C=C— C=C— CH=CH— CH2OH 

Polyporus anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, /. Chem. 
Soc, 1607 (1957). 

204 Deca-czs-24rans-8-diene-4,6-diyn-l-ol, CioHjoO, m.p. <20°, U.V. 

213.5, 230, 237.5, 246.5, 261.5, 276.5, 293.5, 312.5 m^x. 

CH3— CH=CH— C=C— C=C— CH=CH— CH2OH 

Polyporus guttalatus 

J. N. Gardner, E. R. H. Jones, P. R. Leeming and J. S. 
Stephenson, /. Chem. Soc, 691 (1960). 



113 Polyenes and Polyynes, Excluding Polyene Macrolides 

205 10-Hydroxydec-trans-2-ene-4,6-diynoic Acid, C^^H^oO^, colorless 

plates, m.p. 154.5°, U.V. 215, 222 (243) (225), 270, 
285, 303 m;^ in ethanol. 

HOCH2— CH.— CH,— C=C— C^C— CH=CH— COOH 

Polyporus anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, /. Chem. 
Soc, 1607 (1957). 

206 Dimethyl Octa-trans-2,trans-6-dien-4-yne-I,8-dioate, CioHi„04, col- 

orless plates, m.p. 117-119.5°, U.V. (205), 214 (240) 
(278), 292, 307 m/x in ethanol. 

CH3OOC— CH=CH— C=C— CH=CH— COOCH3 

Polyporus anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, /. Chem. 
Soc, 1607 (1957). 

207 Undec-3,5,6-triene-8,10-diynoic Acid, CnHgOa. 

HC^C— C=C— CH=C=CH— CH=CH— CH2— COOH 

Drosophila semivestita 

Marjorie Anchel, Science 126 1229 (1957). 

208 Nemotin, CnHgOo, unstable except in solutions, [ajn^^ +380° (c 

0.3 in ether), U.V. 207, 236, 248, 262, 276 m^x in water. 

HC=C— C=C— CH=C=CH— CH— CH2— CH2— C=0 

I o ' 

and 

209 Nemotinic Acid, CnHioOg, unstable except in solutions, [ajn" 

+320° (c 0.2 in ether), U.V. 208, 237, 249, 263, 277 m^x 
in water. 

CH^C— C=C— CH=C=CH— CH— CH2— CH2— COOH 

I 
OH 

Porta corticola, P. tenuis and another unidentified ba- 
sidiomycete 

Yields of mixed acetylenes from one of the fungi were : 



Pfizer Handbook of Microbial Metabolites 



114 



TABLE I 



Compound 


Concentration in 

the medium 

(mg. per liter) 


Per cent of total 




110 

14 

34 

5 


67.5 




8.5 




21 




3 







J. D. Bu'Lock, E. R. H. Jones and P. R. Leeming, /. Chem. 
Soc, 4270 (1955). (Structure) 

210 Methyl trarzs-10-Hydroxydec-2-ene-4,6,8-triyn-l-oate, CuHgOa, 

needles (dec. -115°), U.V. 245, 256.5, 283, 301, 320.5, 
343.5 rti/x in carbon tetrachloride. 

HOCH,— C^C— C=C— C^C— CH=CH— COOCH3 

Pleurotus ulmarius, Merulius lachrymans 
J. N. Gardner, E. R. H. Jones, P. R. Leeming and J. S. 
Stephenson, J. Chem. Soc, 691 (1960). 

211 trans, trans-Matricaria Ester, CnHj„02, colorless needles, m.p. 

62°, U.V. (234), 246, 258 (296), 314, 333 m^x in ethanol. 

CH3— CH=CH— C^C— C=C— CH=CH— COOCH3 

Polyporus anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, J. Chem. 
Soc, 1607 (1957). 

212 Methyl 10-Hydroxydec-trans-2-ene-4,6-diynoate, CnHioO.^, nearly 

colorless oil, U.V. 215, 223 (243), 258, 273, 287, 305 m^ 
in ethanol. 

HOCHo— CHo— CH.— C=C— C=C— CH=CH— COOCH3 

Polyporus anthracophilus, Merulius lachrymans 
J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, J. Chem. 
Soc, 1607 (1957). 

213 Odyssin, Ci2H,„02, unstable except in solutions, [a]i,'° +360° (c 

0.2 in ethanol), U.V. 210, 237.5, 250, 264, 280 m^. 



CH3 



and 



-C=C— C=C— CH=C=CH— CH—CH,— CH2— C=0 

I O ' 



115 Polyenes and Polyynes, Excluding Polyene Macrolides 

214 Odyssic Acid, Ci2H,oO;i, unstable except in solutions, [aln^" +300° 

(c 0.25 in etiianol), U.V. 211, 238, 250.5, 265, 280.5 m^x. 

CHs— C=C— C=C— CH=C=CH— CH— CH,— CH.— COOH 

OH 

Poria corticola, P. tenuis 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, /. Chem. 
Soc, 1607 (1957). 

215 Dimethyl Deca-2,4,6-triyne-l,10-dioate, Ci2Hi„04, colorless nee- 

dles, m.p. 45°, U.V. 209, 217, 226", 257, 272, 288, 307, 
329 m^ in carbon tetrachloride. 

CH3OOC— C=C— C=C— C=C— CH,— CHo— COOCH3 

Merulius lachrymans 

J. N. Gardner, E. R. H. Jones, P. R. Leeming and J. S. 
Stephenson, /. Chem. Soc, 691 (1960). 

216 Dimethyl Deca-trans-2,trar2S-8-diene-4,6-diyne-l,10-dioate, 

Ci2H,„04, colorless plates, m.p. 104.5-107.5°, U.V. 216, 

269, 298, 317, 339 m^ in ethanol. 

CH3OOC— CH-=CH— C=C— C^C— CH=CH— COOCH3 

Polyporus anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, J. Chem. 
Soc, 1607 (1957). 

217 Dimethyl Dec-trans-2-ene-4,6-diyne-l,10-dioate, C12H12O4, color- 

less crystals, m.p. 56.5-58°, U.V. 214.5, 223 (243) (255), 

270, 285, 303 m,x in ethanol. 

CH3OOC— CH,— CH,— C^C— C=C— CH=CH— COOCH3 

Polyporus anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, J. Chem. 
Soc, 1607 (1957). 

218 Mycomycin, C13H10O0, colorless needles, m.p. 75° (dec. explo- 

sively), [a]rr' -130° (c 0.4 in ethanol), U.V. 256, 267, 
281 rufx in diethyl ether. 

HC=C— C^C— CH=C=CH— CH=CH— CH=CH— CH2— COOH 

Nocardia acidophilus 

Walter D. Celmer and I. A. Solomons, /. Am. Chem. Soc 74 
1870, 3838 (1952). (Structure) 

Edwin A. Johnson and Kenneth L. Burdon, /. Bacteriol. 54 
281 (1947). 



Pfizer Handbook of Microbial Metabolites 1 1 6 

219 Corticrocin, C14H14O4, orange-red, amorphous powder or yellow 
needles and prisms, m.p. subl. 270°, m. 317° (sealed 
tube), U.V. 374, 393, 416 m^a in ethanol. 




Corticeum croceumBies. (= Corticium sulfureum (Fr. ) 
Fr.) 

Yields of about 4% of the mycorrhizal weight have 
been reported. 

Holger Erdtman, Acta Chem. Scand. 2 209 (1948). 
(Isolation and Structure) 

B. L. Shaw and M. C. Whiting, J. Chem. Soc, 3217 (1954). 
(Synthesis) 

B. C. L. Weedon, ibid. 4168 (1954). (Synthesis) 

220 Nemotinic Acid Xyloside, CjsHisO^, [a],."' +237° (c 0.1 ethanol). 
HC=C— C^C— CH=C=CH— CH— CHo— CHo— COOH 

/ o^ 

OH >HO 



HO 



OH 



Basidiomycete B-841 

J. D. Bu'Lock and H. Gregory, Experientia 15 420 (1959). 

22 1 Deca-trans-2-trarzs-8-diene-4,6-diynyl Deca-trans-2, trans-8-diene- 
4,6-diynoate, CooHigOo, colorless crystals, m.p. 124-126°, 
U.V. 213 (233"), 238.5, 246, 259, 277.5, 295, 314, 335 
mfx in ethanol. 

CH3— CH=CH— C=C— C^C— CH=CH— COO— CH2— CH=CH— C=C— C^C— 

CH=CH— CH3 

Polyporus anthracophilus 

J. D. Bu'Lock, E. R. H. Jones and W. B. Turner, J. Chem. 
Soc, 1607 (1957). 



1 1 7 Polyenes and Polyynes, Excluding Polyene Macrolides 

222 Methyl 10-(Deca-fra72S-2,frflns-8-diene-4,6-diyn-l-oyloxy)-dec-trans- 

2-ene-4,6-diynoate, CoiHi^O,, colorless plates, m.p. 91-93°, 
U.V. 223, 246.5, 259", 287, 305, 334 m^u in ethanol. 

CHs— CH=CH— C=C— C=C— CH^CH— COO— CH2— CH2— CHo— C=C— C=C 

— CH=CH— COOCH3 

Polyporus anthracophilus 

J. D. BuLock, E. R. H. Jones and W. B. Turner, 7. Chem. 
Soc, 1607 (1957). 

223 Cortisalin, C^iHoiiO;^, violet-red needles, m.p. dec. >290°, U.V. 

(318),"345 (420), 443 (462) m^a in pyridine. 

HO-Q- 



Corticium salicinum Fries 

A yield of 2.6 g. of crude material was obtained from 
222 g. of fungal fruiting body. 

Jarl Gripenberg, Acta Chem. Scand. 6 580 (1952). 

D. Marshall and M. C. Whiting, /. Chem. Soc, 537 (1957). 
( Synthesis ) 

224 Limocrocin, CosH^^OqN^, a yellow actinomycete pigment. Dark 
red crystals from AcOH m.p. 316° (dec). Dimethyl ester 
of perhydo-deriv., fine, colorless needles, m.p. 146-147°. 
Partial structure: 



O 

\ 
HO— C\^v^^^?\^::r\^:;\^^;;\^~v^^:^v^ ^NHfCsHjONlCOOH 



C 



A demethylcrocetin derivative with the CsHjON probably a 
heterobicycHc residue. Eq. wt. 225 (232). 

Streptomyces limosus (Glycine-glycerol substrate) 
Hans Brockmann and Hans-Ulrich May, Chem. Ber. 88 419 
(1955). 

Hans Brockmann and Gerhard Grothe, Chem. Ber. 86 1110 
(1953). 



Macrocyclic Lactones (Macrolides) 



The macrolide (macrocyclic lactone) antibiotics are an in- 
teresting new class of compounds elaborated by members of the 
order actinomycetales and particularly by the genus strepto- 
myces. The lactone moieties of these molecules resemble the 
partially oxidized and alkylated aliphatic acids characteristic 
of the related mycobacterium genus. A partial listing accord- 
ing to Bergey's Manual of the members of the order actinomy- 
cetales is shown below to clarify these relationships. 

Order. . . Acfinomycefales*^ 



Families. . . Actino- 

mycetaceae 



Strepto- 
mycefaceae 



Myco- Actino- 

bacteriaceae planaceae 



Genera... Nocardia Sfreptomyces* Mycobacterium Actinoplanes 

Actinomyces* Micromonospora Mycococcus Streptosporan- 

Thermoactino- gium 
myces 

A resemblance- to the steroid glycosides, for example strophan- 
thin and oleandrin shown below, also has been noted. ^ 




OH 

Strophanthin 

* In the vernacular usage streptomycete-streptomyces and actino- 
mycete may indicate either order or genus, perhaps more commonly 
the order. 

1 R. B. Woodward, Festschr. Arthur Stoll, 524 (1957). 



119 



Macrocyclic Lactones (Macrolides) 




OCOCH3 



In this regard it is striking that the sugar L-oleandrose occurs 
in both oleandrin and in the macroHde oleandomycin. 

The macrolide antibiotics are most effective against gram- 
positive bacteria. In the introduction to the section on steroids 
and terpenoids, it was mentioned that no true steroids have 
ever been detected conclusively in bacteria. It was noted also 
that certain investigators exploring the utilization of mevalonic 
acid by gram-positive bacteria (especially lactobacilli) found 
that partially oxidized aliphatic substances with more than 15 
carbon atoms were produced.- While these products were not 
thoroughly characterized, the properties as described were rem- 
iniscent of the lactone portions of the macrolides. It also has 
been mentioned elsewhere that the general chemical structure 
and metabolism of the actinomycetales seem to be more closely 
related to that of the bacteria than to that of the fungi, which 
they resemble superficially. From these premises, it is tempting 
to speculate that the macrolide antibiotics may interfere in some 
way with a primitive kind of hormonal or steroid metabolism 
in gram-positive bacteria. In this connection it should be 
noted, however, that the sugar portions of most of the known 
macrolide antibiotics are essential to their antibacterial activity. 
Tylosin and lankamycin may be exceptions. 

Several of the many macrocyclic lactones which have been 
isolated from streptomycete cultures have been well character- 
ized structurally. Complete structures have been reported for 
picromycin, methymycin, neomethymycin, erythromycin, eryth- 
romycin B, erythromycin C, carbomycin (Magnamycin), carbo- 
mycin B, oleandomycin and pimaricin. A considerable amount 
of information has been reported concerning the structures of 
narbomycin, the foromacidins (spiramycins) and the pentaenes 
lagosin and fiUpin.* 

The few cases available for comparison fall into a general 
pattern. This involves the lactone of a long chain aliphatic 

- E. Kodlcek, Abstracts of the Gordon Conference on Vitamins 
and Metabolism, 1958. 
* See addendum. 



Pfizer Handbook of Microbial Metabolites 120 

acid, quite evidently acetate-derived, in conjugation with one or 
more sugar-like moieties. These sugars are uncommon ones, 
and one of them is usually an amino-sugar, desosamine being 
particularly prevalent so far. Several of the incompletely char- 
acterized macrolides, especially those of the polyene type, have 
been reported to contain no nitrogen, however. Among these 
are lagosin, fungichromin, A-246, miamycin and filipin. One 
macrolide, celesticetin,* contains sulfur. Lankamycin also con- 
tains no nitrogen. 

Of all the macroHdes the biosynthesis of erythromycin has 
been investigated most thoroughly. One of the questions to be 
answered was whether the erythronolide moiety is derived from 
acetate or from propionate. A labeling and degradation study 
wdth C^*-containing precursors has shown that propionate or its 
biological equivalent is the true precursor.^ Propionate-C-1 was 
incorporated only into the "methylene" carbon atoms, while 
propionate C-2 was incorporated largely into the tertiary carbon 
atoms and not at all into the carbon-bound methyl groups. Ad- 
ditional evidence against the acetate hypothesis was the fact 
that C^Mabeled formate or C^*-methyl methionine did not label 
the terminal three carbon atom subunit of erythronolide. 

A previous study* had shown that C"-l-labeled propionate 
caused labehng of erythronolide, but not of the sugars desosa- 
mine and cladinose. The reverse was true when the labeled 
precursor was C'*-methyl methionine.^ 

Other evidence which has been published suggests or is con- 
sistent with derivation of erythronolide from propionate." 

A notice has been pubhshed that a labehng study on the 
biogenesis of erythromycin is in progress with the use of pro- 
pionic acid-l-C"-H^' 

It remains to be seen whether or not some of the less highly 

^ John W. Corcoran, Toshi Kaneda and John C. Butte, /. Biol. 
Chem. 235 pc29 (1960). 

* Z. Vanek, J. Majer, A. Babicky, J. Liebster, K. Veres and L. 
Dolezilova, Abstr. IVth Intern. Congr. Biochem., Vienna, 1958; cf. 
Angew. Chem. 71 40 (1959). 

^ Z. Vanek, J. Majer, J. Liebster, K. Veres and L. Dolezilova, 
Symposium on Antibiotics, Prague, 1959. 

^ V. Musilek and V. Sevcik, Naturwissenschaften 45 86 215 (1958); 
idem.. Symposium on Antibiotics, Prague, 1959. 

^ H. Grisebach, H. Achenbach and U. C. Grisebach, Naturwissen- 
schaften 47 206 (1960). 

* See entry 923 for non-macrolide structure. 



121 



Macrocyclic Lactones (Macrolides) 



branched lactones are derived in whole or in part from acetate. 

It is obvious that in each case many modifications of the mac- 
rolide moiety have occurred from the simplest intermediate 
ring which could be envisaged. These include complete or 
partial reduction of carbonyl groups, dehydration of the corre- 
sponding secondary alcohols, epoxidation or reduction of carbon- 
carbon double bonds, oxidation of tertiary carbon atoms, cleav- 
age of epoxides to glycols, etc. Yet, despite the confusing detail, 
the fundamental pattern of oxidation and reduction remains 
apparent, just as it does in many of the metaboUtes of the myco- 
bacteria and corynebacteria. 

It wall be interesting to see how much of the information con- 
cerning the biogenesis of the macrolides can be transposed to 
metabolites of the mycobacteria and corynebacteria and vice 
versa. 

In the cases of picromycin, methymycin, erythromycin, nar- 
bomycin and oleandomycin it is possible to follow the course of 
alternate oxidation throughout the lactone rings wdth remark- 
able regularity, the hypothetical intermediate being, apparently, 
a single continuous chain, unbranched except for the methyl 
groups. In the cases of carbomycin and pimaricin, anomalies 
occur. These could be explained by a junction of shorter 
chains, perhaps as shown below, in a manner similar to the for- 
mation of corynomycolic acid by the coupling of 2 moles of pal- 
mitic acid: 





Pimaricin 



OCOCH3 
Lactone Portion of Carbomycin 



Another suggestion has been made in the case of carbomycin,^ 
namely that a protocarbomycin may occur which later rear- 
ranges by a glycol-aldehyde shift: 



Pfizer Handbook of Microbial Metabolites 



122 




C— O— H 



^ C-O-H 



I 

c=o 

1 

H 



CH3 O OCOCH3 

Proposed Precursor of the Proposed Glycol-aldehyde 

Lactone Portion of Corbomycin Rearrangement at 7,8-Positions 

Such a precursor is the more plausible because it would have an 
1 8-membered carbon atom chain and a C-19 carbon skeleton, 
the same as that of the known tuberculostearic acid, even in- 
cluding the stereochemistry of the branching methyl group. 

Streptomyces species produce many antifungal antibiotics 
which have in common chains of conjugated olefinic bonds. By 
means of the ultraviolet absorption spectra it is possible to clas- 
sify them according to the length of the conjugated chain. 
Generally these substances are rather intractable with low solu- 
bilities and indefinite melting points. 

A structure has been proposed for pimaricin, a tetraene. 
Whether or not this structure proves to be entirely correct, there 
is evidence from several sources that at least certain of these 
substances are macrocyclic lactones. 

So many of these compounds have been reported lately that 
any listing is likely to be incomplete. The following table must 
include most of them, however, grouped by number of conju- 
gated olefinic bonds. 



TABLE I 



Tetraene 


Hexaene 


Pentaene 


Heptaene 


Nystatin (Fungicidin) 


Fradicin 


Eurocidin 


Amphotericin B 


Rimocidin 


Flavacid 


Fungichromatin 


Candidin 


Pimaricin 


Mediocidin 


Fungichromin 


Candicidin 


Amphotericin A 


Endomycin B (Helixin B) 


Filipin 


Candimycin 


Protocidin 




PA- 153 


Ayfactin 


Chromin 




Pentamycin 


Ascosin 


Antimycoin 






Trichomycin 


Sistomycosin 






PA- 150 


Endomycin A (Helixin A) 






Antibiotic 1968 


Etruscomycin 








PA- 166 








Tennecetin 








Flavofungin 









123 



Polyene Macrolides 



Various other substances, e.g. the etamycin, valinomycin and 
actinomycin types of antibiotics, could be classed as macrolides 
since they all contain large rings in which lactone groups par- 
ticipate. 

a. POLYENE MACROLIDES 

225 Flavofungin, C^„H4s09 Dihydrate. 

A polyene macrolide containing 7 acetylatable hydroxyl 
groups, 5 hydrogenatable carbon-carbon double bonds of 
which at least 4 are conjugated, contains no alicyclic ring, 
has at least 2 and probably 3 C — CH^. Ozonolysis indi- 
cates a CH;^(C,jHii) group. The most important struc- 
tural elements are: 



— CH=C- 



<=CH- 



CH3 CH3 



-(CH=CH)3— C=0, CH3(C6H„) 
O 



CloHi3(OH)7 



Shown to be distinct from pimaricin, nystatin, amphoteri- 
cin-B, fungichromin, lagosin, filipin and fumagillin. 

A streptomycete 

R. Bognar, Angew. Chem. 72 139 (1960). 

226 Pimaricin ( Myprozine ) , C34H49O14N, colorless crystals, m.p. 
200° (dec), U.V. 279, 290, 303, 318 m^ in methanol. 
Proposed structure: 

CH2OH 

I 
OH O O CHOH 




Streptomyces natalensis n. sp. 

A. P. Struyk, I. Hoette, G. Drost, J. M. Waisvisz, T. Van Eek 
and J. C. Hoogerheide, "Antibiotics Annual 1957-1958," Med- 
ical Encyclopedia, Inc., New York, p. 878. 

James B. Patrick, Richard P. Williams and John S. Webb, 
J. Am. Chem. Soc. 80 6689 (1958). (Structure) 



Pfizer Handbook of Microbial Metabolites 1 24 

227 PA-166, C35H53O14N (proposed), colorless powder, m.p. gradual 

dec. up to 260°, [a]n'' +275° (c 0.2 in pyridine). 

An amphoteric tetraene. U.V. maxima: 291, 304, 319 
in aqueous methanol. Positive ninhydrin, 2,4-DNPH and 
Fehling's tests. Three C-methyl groups. 

Streptomyces n. sp. 

B. K. Koe, F. W. Tanner, Jr., K. V. Rao, B. A. Sobin and 
W. D. Celmer, "Antibiotics Annual 1957-1958," Medical En- 
cyclopedia, Inc., New York, p. 897. 

228 Etruscomycin, CagH^TOi^N, white crystals, [a]D^° +296° (c 1 in 

pyridine ) . 

A tetraene antibiotic. I.R. peaks at: 2.91, 3.38, 5.83, 
6.30, 9.44, 9.55, 11.85;x. U.V. peaks at: 290, 300, 316 m^x. 

Streptomyces lucensis n. sp. 

F. Arcamone, C. Bertazzoli, G. Canevazzi, A. DiMarco, M. 
Ghione and A. Grein, Giorn. Microbiol. 4 119 (1957). 

229 Lagosin (Antibiotic A-246), C41H66-70O14. m.p. ^235° (dec), 

[a]ir" -160° (c 0.2 in methanol). 

An antifungal pentaene macrolide antibiotic with the 
following partial structure : * 

n— CsHnCHOH 

I 

O— CO— CH— c— c— c— c— c— c— c— c— c— c— 

I 

CH3— CH— CH— (CH=CH)4— CH=C CH C— 



> C6H30-34O9 



OH CH3 OH OH 

Streptomyces sp. 

M. L. Dhar, V. Thaller and M. C. Whiting, Proc. Chem. Soc, 
148 (1958)._ 

M. L. Dhar, V. Thaller, M. C. Whiting, Ragnar Ryhage, 
Stina Stalberg-Stenhagen and Einar Stenhagen, ibid., 154 
(1959). (Structure) 

S. Ball, Christine J. Bessell and Aileen Mortimer, J. Gen. 
Microbiol. 17 96 (1957). (Isolation) 

230 Nystatin (Fungicidin, Mycostatin) C46H77O19N (tentative), yel- 
low powder, m.p. dec. above 160°, but no definite m.p., 
[aln'^ +10° (in glacial acetic acid). 

An amphoteric tetraene. U.V. maxima at: 280, 291, 
304, 318 m/A. Contains a mycosamine moiety: 

NH2 
HO I OH 



O: 



HO CH3 

* See addendum. 



125 Polyene Macrolides 

Streptomyces noursei 

Elizabeth L. Hazen and Rachel Brown, Proc. Soc. Expl. 
Biol. Med. 76 93 (1951). 

James D. Dutcher, Gerald Boyack and Sidney Fox, "Anti- 
biotics Annual 1953-1954," Medical Encyclopedia, Inc., New 
York, p. 191. 

David R. Walter, James D. Dutcher and O. Wintersteiner, 
;. Am. Chem. Soc. 79 5076 (1957). (Structure) 

231 Rimocidin (Sulfate heptahydrate), large fragile plates, m.p. 

^151° (dec.), [aW (sulfate) +75° (c 1 in methanol). 

An amphoteric tetraene. U.V. maxima at: 279, 291, 
304, 318 m/x. Analysis (hydrated sulfate): C 57.65, H 
7.82, N 1.81, S 2.03. 

Streptomyces rimosus 

J. W. Davlsson, F. W. Tanner, Jr., A. C. Finlay and I. A. 
Solomons, Antibiotics and Chemotherapy 1 289 (1951). 

232 Protocidin, m.p. dec. from 120°. 

A polyene antifungal agent. U.V. maxima 277, 290, 
303 and 318 mpi. Reduces KMn04. Green Fehling. Neg- 
ative biuret, Sakaguchi, Molisch, ninhydrin, anthrone, 
FeCL. 

Streptomyces sp. 

The yield was about 100 mg. per liter. 

Jean Marie Sakimoto, J. Antibiotics (Japan) lOA 128 
(1957). 

233 Amphotericin-A, m.p. gradual dec. above 153°, [(x]o^^"^ +32° (in 

acid dimethylformamide). 

An amphoteric tetraene. U.V. maxima: 291, 305, 320 
mix. Analysis: C 60.32, H 8.39, N 1.72. 

Streptomyces sp. 

J. Vandeputte, J. L. Wachtel and E. T. Stiller, "Antibiotics 
Annual 1955-1956," Medical Encyclopedia, Inc., New York, 
p. 587. 

234 Sistomycosin, bufp or light yellow microcrystals, m.p. --'230° 

(browning from 130°). 

A neutral tetraene. U.V. maxima: 218, 292.5, 306, 
320.5 m^ in aqueous solution. Positive Benedict and 
Molisch tests. 

Streptomyces viridosporus n. sp. 

J. Ehrlich, M. Knudsen and Q. Bartz, Canadian Patent 
514,894 (1955). 

235 Endomycin A (Helixin A), yellow-brown powder. 

An acidic tetraene. U.V. maxima at 292, 301, 319 mix. 
Streptom.yces hygroscopicus (S. endus) 
A yield of 11.7 g. of mixed endomycins from about 15 
liters of broth has been reported. 



Pfizer Handbook of Microbial Metabolites 126 

L. C. Vining and W. A. Taber, Can. ]. Chem. 35 1461 
(1957). 

David Gottlieb, P. K. Bhattacharyya, H. E. Carter and H. W. 
Anderson, Phytopathology 41 393 (1951). (Isolation) 

Curt Leben, G. J. Stessel and G. W. Keitt, Mycologia 44 159 
(1952). 

R. R. Smeby, Curt Leben, G. W. Keitt and F. M. Strong, 
Phytopathology 42 506 (1952). 

236 Tennecetin, yellow amorphous powder. 

A tetraene antibiotic. U.V. absorption peaks at 288, 
300-302, and 315-318 m^. 

Streptomyces chattanoogensis 

James Burns and D. Frank Holtman, Antibiotics and Chem- 
otherapy 9 398 (1959). 

237 Antimycoin, organic acid, U.V. maxima: 291, 304—305, 318 

mjx in ethanol. Similar to fungicidin. (A tetraene) 
Streptomyces aureus Waksman and Curtis 
Carl P. Schaffner, Irwin D. Steinman, Robert S. Safferman 

and Hubert Lechevalier, "Antibiotics Annual 1957-1958," 

Medical Encyclopedia, Inc., New York, pp. 5869-5873. 

Frederick Raubitscheck, Robert F. Acker and Selman A. 

Waksman Antibiotics and Chemotherapy 2 179 (1952). 

238 Filipin, C:^2H-,40io, fine yellow needles, m.p. 195-205° (dec.) (s. 

147°), [a],," -148.3° (c 0.89 in methanol). 

A neutral pentaene. U.V. maxima at 322, 338, 355 m^. 
Contains 7-8 acetylatable non-vicinal hydroxyl groups and 
3-4 C— CH. groups. 

Possible partial structure : * 

CH3 OH OH 

-t I I I 

CH3— C— C— C=C— (C=C)4— c— c — c 

OH I C 

o I 

o=c— C— C— C— C— C— C— C— C— OH 

n— CsHu— C— OH 

Streptomyces fdipinensis n. sp. 

Geo. B. Whitfield, Thomas D. Brock, Alfred Ammann, 
David Gottlieb and Herbert E. Carter, ]. Am. Chem. Soc. 77 
4799 (1955). (Isolation) 

Alfred Ammann, David Gottlieb, Thomas D. Brock, Her- 

* See addendum. 



127 Polyene Macrolides 

bert E. Carter and George B. Whitfield, Phytopathology 45 559 
(1955). 

Belig Berkoz and Carl Djerassi, Proc. Chem. Soc, 316 
(1959). (Structure) 

239 Fungichromin, C;{.-,H,i,,0,;!, pale yellow crystals, m.p. 205-210°. 

A pentaene. U.V. maxima: 322.5, 338.5, 356.5 m^. 
The following moiety has been obtained by alkaline hy- 
drolysis followed by periodate oxidation: 

OHC— C=CH(CH=CH)4— CHO 

CH:, 

Streptomyces cellidosae 

A similar substance, fungichromatin, occurred in the 
same culture. 

Alfred A. Tytell, Frank J. McCarthy, W. P. Fisher, Wil- 
liam A. Balhofer and Jesse Charney, "Antibiotics Annual 
1954-1955," Medical Encyclopedia, Inc., New York, p. 716. 

Arthur C. Cope and Herbert E. Johnson, /. Am. Chem. Soc. 
80 1504 (1958). 

240 PA-153, C:i7H,;,Oi4N (proposed), colorless powder, m.p. gradual 

dec. up to 260° ( triethylamine salt dec. 126-129°), [x]^-^ 
-f398° (c 0.2 in pyridine). 

An amphoteric pentaene. U.V. maxima: 303, 317, 
332, 349 in aqueous methanol. Positive ninhydrin, 2,4- 
DNPH and Fehlings tests. Three C-methyl groups. 

Streptomyces n. sp. 

B. K. Koe, F. W. Tanner, Jr., K. V. Rao, B. A. Sobin and 
W. D. Celmer, "Antibiotics Annual 1957-1958," Medical En- 
cyclopedia, Inc., New York, p. 897. 

241 Pentamycin, pale yellow needles, m.p. 237° (dec). 

An antifungal pentaene antibiotic resembling filipin in 
some properties. U.V. maxima at: 322, 338, 356 m^. 
Contains only C, H, O. 

About 60 g. of fairly pure material were obtained from 
100 liters of culture (mycelium). 

Streptomyces penticus 

Sumio Umezawa and Yoshiaki Tanaka, /. Antibiotics (Ja- 
pan) llA 26 (1958). 

242 Eurocidin. 

A pentaene. U.V. maxima: 318, 333, 351 m^x. 



Pfizer Handbook of Microbial Metabolites 128 

Streptomyces alboreticuli n. sp. 

Yashiro Okami, Ryazo Utahara, Shashiro Nakamura and 
Hamao Umezawa, J. Antibiotics (Japan) 7A 98 (1954). 

Ryozo Utahara, Yashiro Okami, Shashiro Nakamura and 
Hamao Umezawa, ibid. 7A 120 (1954). 

243 Fradicin, C3„H3404N4, pale greenish yellow crystals, m.p. dark- 

ens without melting 180-300°, [alo'' +65° (c 1 dioxane). 

Weakly basic hexaene. U.V. maxima: 290-295. Two 
methoxyls. 

Streptomyces fradiae 

E. Augustus Swart, Antonio H. Romano and Selman A. 
Waksman, Proc. Soc. Exptl. Biol. Med. 73 376 (1950). 

Richard J. Hickey and Phil Harter Hidy, Science 113 361 
(1951). 

244 Flavacid, pale yellow microcrystalline powder, m.p. 102-105° 

(dec). 

A weakly acidic hexaene. U.V. maxima: 340, 360, 
380 mix. A tetraene with peaks at 293, 306 and 324 is 
also present. 

A streptomycete resembling S. fiaviis 

Isao Takahashi, /. Antibiotics (Japan) 6A 117 (1953). 

L. C. Vining and W. A. Taber, Can. J. Chem. 35 1461 
(1957). 

245 Mediocidin, yellow amorphous powder. 

A hexaene. U.V. maxima: 340, 357, 378 m/x. A 
tetraene, probably identical with that in the flavacid com- 
plex, is also present. U.V. maxima: 293, 306, 324. 

Streptomyces mediocidicus, n. sp. 

Ryazo Utahara, Yoshiro Okami, Shashiro Nakamura and 
Hamao Umezawa, J. Antibiotics (Japan) 7A 120 (1954). 

L. C. Vining and W. A. Taber, Can. J. Chem. 35 1461 
(1957). 

246 Endomycin B (Helixin B), yellow-brown powder. 

An acidic hexaene. For U.V. spectrum see first refer- 
ence below. 

Streptomyces hygroscopicus (S. endus) 

L. C. Vining and W. A. Taber, Can. J. Chem. 35 1461 
(1957). 

David Gottlieb, P. K. Bhattacharyya, H. E. Carter and H. W. 
Anderson, Phytopathology 41 393 (1951). (Isolation) 

Curt Leben, G. J. Stessel and G. W. Keitt, Mycologia 44 159 
(1952). 

R. R. Smeby, Curt Leben, G. W. Keitt and F. M. Strong, 
Phytopathology 42 506 (1952). 



129 Polyene Macrolides 

247 Helixins. 

A complex of three or four compounds. Helixin B is 
identical with endomycin B. 

Streptomyces sp. 

Curt Leben, G. J. Stessel and G. W. Keitt, Mycologia 44 159 
(1952). 

248 Amphotericin-B, doHy^OooN (tentative) deep yellow prisms or 

needles from dimethylformamide, m.p: gradual dec. above 
170°, [ajp +333° (in acid dimethylformamide). 

An amphoteric heptaene, U.V. maxima at: 364, 383, 
408 m^. Contains a mycosamine moiety: 



HO V"' OH 


HO CH3 



Streptomyces nodosiis 

J. Vandeputte, J. L. Wachtel and E. T. Stiller, "Antibiotics 
Annual 1955-1956," Medical Encyclopedia, Inc., New York, 
p. 587. (Isolation) 

David R. Walters, James D. Dutcher and O. Wintersteiner, 
J. Am. Chem. Soc. 79 5076 (1957). (Structure) 

249 Zaomycin, m.p. 242-246° (dec). 

An amphoteric antibiotic said to resemble amphotericin. 
Positive ninhydrin, Millon, biuret, FeClo tests. Negative 
Fehling and Liebermann reactions. 

Streptomycin zaomyceticus 

Yorio Hinuma, J. Antibiotics (Japan) 7A 134 (1954). 

250 PA-150, Cr,4H820i8N2 (proposed), yellow powder, m.p. gradual 

dec. up to 260°, [ajrr' +294° (c 0.2 in pyridine). 

An amphoteric heptaene. U.V. maxima: 340, 358, 
377, 397 m^ in aqueous methanol. Positive 2,4-DNPH 
and Fehlings tests. Four C-methyl groups. 

Streptomyces n. sp. 

B. K. Koe, F. W. Tanner, Jr., K. V. Rao, B. A. Sobin and 
W. D. Celmer, "Antibiotics Annual 1957-1958," Medical En- 
cyclopedia, Inc., New York, p. 897. 

251 Trichomycin, yellow powder, m.p. 155° (dec). 

A heptaene. U.V. maxima: 286, 346, 364, 384, 405 
rufx. May be a mixture of two heptaenes. 
Streptomyces hachijoensis n. sp. 



Pfizer Handbook of Microbial Metabolites 130 

Seigo Hosoya, Nobuhiko Komatsu, Momoe Soeda and Yoko 
Sonoda, Japan J. Exptl. Med. 22 505 (1952). 

Seigo Hosoya, Nobuhiko Komatsu, Momoe Soeda, Tatsuro 
Yuwaguchi and Yoko Sonoda, 7. Antibiotics (Japan) 5 564 
(1952). 

252 Candidin, yellow powder. 

Acidic heptaene. U.V. maxima: (Na salt) 234, 282, 
345, 360, 383, 405 m^i in aqueous solution. The free acid 
lacks the 345 peak. Contains nitrogen and gives positive 
ketone tests. 

Streptomyces viridofiavus 

Willard A. Taber, Leo C. Vining and Selman A. Waksman, 
Antibiotics and Chemotherapy 4 455 (1954). 

Leo C. Vining, Willard A. Taber and Francis J. Gregory, 
"Antibiotics Annual 1954— 1955, ' Medical Encyclopedia, Inc., 
New York, p. 980. 

Candicidins. 

Heptaenes. U.V. maxima: 

253 Candicidin A: 360, 380, 403 m/x. 

254 Candicidin B: 362, 381, 404 m^. 

255 Candicidin C: 358, 379, 402 m/x. 

Streptomyces griseus, other Streptomyces spp. 
Hubert A. Lechevalier, R. F. Acker, C. T. Corke, C. M. 
Haenseler and S. A. Waksman, Mycologia 45 155 (1953). 

256 Ascosin, yellow-orange powder. 

A weakly acidic heptaene. U.V. maxima: 234, 288, 
340, 357, 376, 398 m/x in methanol. 

Streptomyces canescus 

Richard J. Hickey, Cyril J. Corum, Phil H. Hidy, I. Ray 
Cohen, Urs F. B. Nager and Eleonore Kropp, Antibiotics and 
Chemotherapy 2 472 (1952). 

Isadore R. Cohen, U. S. Patent 2,723,216, (1955). 



h. OTHER MACROLIDES 

257 Nitrosporin, C^oH^eOfiNo, colorless crystals, m.p. 130-140° 
(dec). Crystals brown on exposure to air. 

A basic substance, apparently a macrolide. 

Streptomyces nitrosporeus 

Hamao Umezawa and Tomio Takeuchi, /. Antibiotics (Ja- 
pan) 5 270 (1952). 



131 



Other Macrolides 



258 Celesticetin I, amphoteric, crystaUine and dextrorotatory, 

Co4H3^.4„Ot,N2S (suggested empirical formula). Oxalate 
and Salicylate water soluble. Oxalate m.p. 149-154°; 
Salicylate m.p. 139° (tabular monoclinic crystals). 

Erythromycin-hke. (See entry 923, however) 

Positive tests — FeCla, Molisch, Ekkert 

White ppt. — Bro water, Millon's Reagent, HgCL 

Negative tests — AgNOg, PbAc, Benedict, ninhydrin, io- 
doform, nitroprusside (becomes + after standing several 
days in 6 N hydrochloric acid) 

No immediate reaction with Br^ — CCI4. 

Streptomyces caelestis 

Herman Hoeksema, Glen F. Crum, William H. DeVries, 
Antibiotics Annual 2 837-841 (1954-1955). (Isolation and 
purification ) 

259 Amaromycin, C^.-.H^^OyN (proposed), colorless prisms, m.p. 

164.5°, [a]n'' +6.19° (c 1 in ethanol). 

Basic substance, analysis: C 63.66, H 8.73, N 3.0. 
Negative FeCl-^, biuret, ninhydrin, Sakaguchi, Schiff. 
Positive Tollens, Fehlings. Precipitated by Reinecke's 
salt. Probably a macrolide. 

Streptomyces flavochromogenes 

Toju Hata, Yashimoto Sano, Hideo Tatsuta, Ryazo Suga- 
wara, Akihiro Matsumae and Kokichi Kanamori, /. Antibiotics 
(Japan) 8A 9 (1955). 



260 



261 



PA-133 A, C2r,H430eN, colorless amorphous solid, [a]n'' +39.6° 
(c 0.5 in methanol). 

A macrolide antibiotic. 

Streptoviyces sp. 

K. Murai, B. A. Sobin, W. D. Celmer and F. W. Tanner, 
Antibiotics and Chemotherapy 9 485 (1959). 

Methymycin, C05H43O7N, colorless prisms 
1 Q70 rnn'io\ [1 23 



197° (203°), [a],,-' +61' 



, needles, m.p. 195— 
(in methanol). 




Desosamine 



Pfizer Handbook of Microbial Metabolites 



132 



A streptomycete 

Carl Djerassi and John A. Zderic, J. Am. Chem. Soc. 78 
2907 (1956). (Structure) 

Milton N. Donin, Joseph Pagano, James D. Dutcher and 
Clara M. McKee, "Antibiotics Annual 1953-1954," Medical 
Encyclopedia, Inc., New York, p. 179. (Isolation) 

262 Neomethymycin, Co,-,H4..07N, colorless crystals, m.p. 156°, [aln^^ 
+93° (in chloroform). 



XH3 

-P. 

OH N— CH3 



CH3 
CH3 



CH3 
O 



CH3 



Desosamine 



OH 



CH3 CH3 

Same streptomycete which produces Methymycin 
Carl Djerassi and O. Halpern, J. Am. Chem. Soc. 79 2022 
(1957). (Structure) 

J. Vandeputte, unpublished. (Isolation) 

263 Picromycin, C25H4:^07N, colorless crystals, m.p. 169.5°, [alo^" 
-33.5° (c 2.07 in chloroform). 



(D 



Picrocin 
esosamine) 



CH3 



CH3— N 
CH3 



CH3 

— o' 



CH3 



CHa 
O 



OH 



/ 



HO CH3 



CH3 



Streptomyces felleus n. sp. 

Hans Brockmann and Rudolf Oster, Chem. Ber. 90 605 
(1957). (Partial structure) 

R. Anliker and K. Gubler, Helv. Chim. Acta 40 119 (1957). 
(Structure) 

Hans Brockmann and Willfried Henkel, Chem,. Ber. 84 284 
(1951). (Isolation) 

Ibid., Naturwissenschaften 37 138 (1950). (Isolation) 



133 Other Macrolides 

264 PA-133-B, Cor.Hj-OioN, colorless crystals, m.p. 99.8-101°, [ajn'' 

+ 22.5° (c 0.5 in methanol). 

A macrolide antibiotic. 

Streptoviyces sp. 

K. Mural, B. A. Sobin, W. D. Celmer and F. W. Tanner, 
Antibiotics and Chemotherapy 9 485 (1959). 

265 Griseomycin (Lomycln) (Hydrochloride) C25H4g08NCl, white 

powder, m.p. 76-80° (dec), [a]n'' +32° (c 1 in chloro- 
form). 

Precipitated by Reinecke salt, bromine water, picric 
acid. Thought to be a macrolide. 

Streptomyces griseolus 

P. J. Van Dijck, H. P. Van de Voorde and P. DeSomer, Anti- 
biotics and Chemotherapy 3 1243 (1953). 

Ibid. Belgian Patent 522,647 (1954). 

266 Proactinomycin A, C27H47OSN (proposed), colorless crystals, 

m.p. 168°. 

267 Proactinomycin B, Co^H4c,OsN (proposed), colorless crystals, 

m.p. 83-87°. 

268 Proactinomycin C, C24H41O6N (proposed), amorphous. 

Basic substances, precipitated by Reineckes salt, picric 
or flavianic acids, etc. Probably macrolides. 

Nocardia gardneri 

A. D. Gardner and E. Chain, Brit. J. Exptl. Path. 23 123 
(1942). 

R. Q. Marston, ibid. 30 398 (1949). (Isolation) 

Antimycins (Antipiriculins)* 

269 Antimycin A^, C28H40O9N2, colorless crystals, m.p. 149-150°, 

[ix]t>'^' +76 (c 1 in chloroform). 

270 Antimycin Aoa, Cor,H3609N2, colorless crystals, m.p. 143-149°, 

271 Antimycin A^^ (may be isomeric with A2a), colorless crystals, 

m.p. 168°. 

272 Antimycin A3 (Blastmycin), C20H3BO9N2, colorless crystals, m.p. 

170.5-171.5°, [x]rr'= +64.3° (c 1 in chloroform). 

* The antimycins might also be classified as depslpeptldes (pepto- 
hdes). 



Pfizer Handbook of Microbial Metabolites 134 



273 Antimycin A4, oily. 



CHj 



O 



O CH3 

NH OH \ / 

/ CH CH 

OHC ^ V r^ ^^ R = n— CeHia in Ai. 

CH3 ^ '\ R = n — C4H9 in A3. 

O 

At least seven streptomyces species produce antimy- 
cins, including S. kitazawaensis Harada et Tanaka nov. 
sp. and S. blastmyceticus. The former organism also pro- 
duces carzinocidin. Blastmycin is identical with anti- 
mycin A;(. Virosin is probably a mixture of antimycin 
components. Certain antimycin-producing cultures also 
contain actinomycin B. 

Wen-chik Liu and F. M. Strong, J. Am. Chem. Soc. 81 4387 
(1959). 

Wen-chik Liu, E. E. Van Tamelen and F. M. Strong, ibid. 
82 1652 (1960). (Degradations, etc.) 

F. M. Strong, J. P. Dickie, M. E. Loomans, E. E. Van 
Tamelen and R. S. Dewey, ibid. 82 1513 (1960). (Structure) 

Bryant R. Dunshee, Curt Leben, G. W. Keitt and F. M. 
Strong, ibid. 71 2436 (1949). (Isolation) 

Yoshio Sakagami, Setsuo Takeuchi, Hiroshi Yonehara, 
Heiichi Sakai and Matso Takashima, J. Antibiotics (Japan) 
9A 1 (1956). 

Kiyoshi Nakayama, Fukusaburo Okamoto and Yujiro 
Harada, ibid. 9A 63 (1956). 

Yujiro Harada, Keizo Uzu and Masaru Asai, ibid. IIA 32 
(1958). 

Hiroshi Yonehara and Setsuo Takeuchi, ibid. IIA 122 
(1958). (Proposed structure) 

Kiyoshi Watanabe, Tsutomo Tanaka, Keiko Fukuhara, 
Norisama Miyairi, Hiroshi Yonehara and Hamao Umezawa, 
ibid. lOA 39 (1957). 

F. M. Strong, "Topics in Microbial Chemistry" (Squibb 
Lectures on the Chemistry of Microbial Products), John Wiley 
and Sons, Inc., New York, 1956, pp. 1-44. (A review to that 
date) 



135 



Other Macrolides 



274 



Naibomycin, C^-sH^oOyN, colorless crystals, 
[ai- +68.5° (c 1.35 in chloroform). 



m.p. 113.5-115°, 





f CH3 CH3 




Desosamine 


1 .OH 




CH3 





CH3 1 


II CH 


CH3 


CH3 



1 


CHo 

1 






CH3 CH3 



Streptomyces narboensis n. sp. 

R. Corbaz, L. Ettlinger, E. Gaumann, W. Keller-Schierlein, 
F. Kradolfer, E. Kyburz, L. Neipp, V. Prelog, P. Reusser, and 
H. Zahner. Helv. Chim. Acta 38 935 (1955). 

R. Anliker, D. D. Dvornik, K. Gubler, H. Heusser and V. 
Prelog, ibid. 39 1785 (1956). 

V. Prelog, A. M. Gold, G. Talbot and A. Zamojskl. (To be 
published) 

275 Leucomycin, C:i3,;^s;H54.p,jO,,_,;{N, colorless crystals, m.p. 124- 
125.5°, [air,-" -67.1° (c 1 in ethanol). 

Leucomycin appears to be a macrolide antibiotic* 

Streptomyces kitasatoensis n. sp. 

Toju Hata, Yoshimoto Sano, Natsuo Ohki, Yasuhiku Yoko- 
yama, Akihiro Matsumae and Shinya Ito, /. Antibiotics (Ja- 
pan) 6A 87 (1953). 

Yoshimoto Sano, Tadashi Hoshi and Toju Hata, ibid. 7A 
88 (1954). 

Yoshimoto Sano, ibid. 7A 93 (1954). 



* See addendum. 



Pfizer Handbook of Microbial Metabolites 



136 



276 Oleandomycin (PA-105), Ca^.H^iOjoN, colorless prisms, m.p. 110' 
(dec), [a]i)"^ —65° (c 1 in methanol). 



Oleandrose 





f CH3 CH3 1 




\ / 




N OCH3 _ 




1 OH 1 OH 


Desosamine- 


Xy Xy 




CH3 CH3 


CH3 1 CH3 


CH3 ^^ 
CH2 .^ OH \ 


^o^CVx ° 




1. '.. CH3 



CH3 CHa 

Streptomyces antibioticus 

B. A. Sobin, A. R. English and W. D. Celmer, 'Antibiotics 
Annual 1954-1955," Medical Encyclopedia, Inc., New York, 
p. 827. 

W. D. Celmer, H. Els and K. Murai, "Antibiotics Annual 
1957-1958," Medical Encyclopedia, Inc., New York, p. 476. 

Hans Els, Walter D. Celmer and Kotaro Murai, J. Am. 
Chem. Soc. 80 3777 (1958). 

W. D. Celmer, "Antibiotics Annual 1958-1959," Medical 
Encyclopedia, Inc., New York, p. 277. (Biochemical correla- 
tions ) 

F. A. Hochstein, H. Els, W. D. Celmer, B. L. Shapiro and 
R. B. Woodward, /. Am. Chem. Soc. 82 3225 (1960). (Struc- 
ture) 

277 Erythromycin C, C;^,;H,;-,0]3N, white needles, m.p. 121-125°. 

Erythromycin C differs from erythromycin only in the 
neutral sugar moiety, so that the following partial struc- 
ture can be written: 



Desosamine 




O O— C7H13O3 



OH CH3 

OH O O 
OH 




CH3 CH3 CH3 



137 



Other Macrolides 



Streptoviyces erythreus 

Paul F. Wiley, Richard Gale, C. W. Pettinga and Koert 
Gerzon, /. Am. Chetn. Soc. 79 6074 (1957). (Structure and 
isolation) 

278 Erytliromycin B, CinHcyOj.^N, colorless crystals, m.p. 198°, [alo"^ 
-78° (c 2 in ethanol). 



Desosamine ' 




Cladinose 



CHs Cms CHs 

Streptomyces erythreus 

Paul F. Wiley, Max V. Sigal, Jr., Allidene Weaver, Rosema- 
rie Monahan and Koert Gerzon, /. Am. Chem. Soc. 79 6070 
(1957). (Structure) 

C. W. Pettinga, W. M. Stark and F. R. Van Abeele, ibid. 76 
569 (1954). (Isolation) 

279 Erythromycin (Ilotycin, Erythrocin), CojHgyOiaN, white needles, 
m.p. 136-140°, [aW -78° (c 1.99 in alcohol). 



Desosamine" 




Cladinose 



Erythronolide 



Streptomyces erythreus 

R. K. Clark, Jr. Antibiotics and Chemotherapy 3 663 
(1953). (Isolation) 

Paul F. Wiley, Koert Gerzon, Edwin H. Flynn, Max V. 
Sigal, Jr., Allidene Weaver, U. Carol Quarck, Robert R. Chau- 



Pfizer Handbook of Microbial Metabolites 



138 



vette and Rosemarie Monahan, J. Am. Chem. Soc. 79 6062 

(1957). (Structure) 

280 PA-108, C,.isH6:hOi4N, colorless solid, m.p. 121-123°, [^W -36.8° 

(c 1 in chloroform). 

A macrolide antibiotic. 

Streptoviyces sp. 

K. Murai, B. A. Sobin, W. D. Celmer and F. W. Tanner, 
Antibiotics and Chemotherapy, 9 485 (1959). 

281 PA-148, C3sH6r,Oi-,N, colorless amorphous solid, m.p. 115-118°, 

[a]v-^ -69.3° (c 0.5 in methanol). 

A macrolide antibiotic. 

Streptomyces sp. 

K. Murai, B. A. Sobin, W. D. Celmer and F. W. Tanner, 
Antibiotics and Chemotherapy, 9 485 (1959). 

282 Carbomycin B, C4.Hf570i-,N, colorless plates, m.p. 141-144° 

(dec), Hydro'chloride 164-166° (dec), [oc],,-' -35° (c 2.0 
in chloroform). 



CH3 




OCOCH2CH(CH3)2 
CH3 



Isovaleryl 
Mycarose 



CH 



Streptomyces halstedii 

F. A. Hochstein and Kotaro Murai, /. Am. Chem. Soc. 76 
5080 (1954). (Isolation) 

R. B. Woodward, Angexv. Chem. 69 50 (1957). (Struc- 
ture) 

283 Carbomycin (Magnamycin), C42H(;70,,jN, colorless laths, m.p. 
212-214° (dec), [aW -58.6° (c 1 in chloroform). 




Carimbose 

CHO CH 

J /O 



CH3 



OCOCH2CH(CH3)2 
CH3 

Isovaleryl 
Mycarose 




OH 



N(CH3)2 



OCOCH3 



Mycaminose 



139 Other Macrolides 

Streptomyces halstedii, S. alboreticuli 
R. B. Woodward, Angew. Chem. 69 50 (1957). (Structure) 
Richard L. Wagner, F. A. Hochstein, Kotaro Murai, N. Mes- 
sina and Peter B. Regna, J. Am. Chem. Soc. 75 4684 (1953). 
(Isolation) 

284 Tertiomycin A, C4oH490ieN, white needles, m.p. 215-217° (s. 

208") (dec.)" [a],/' -49° (c 1 in chloroform) [aW^ -47° 

(c 1.0 in ethanol). 

A macrolide antibiotic. Carbomycin produced also by 

S. alboreticuli. 

Streptomyces euroddicus, S. alboreticuli 

Teisuke Osato, Masahiro Ueda, Setsuko Fukuyama, Koki 

Yagishita, Yoshiro Okami and Hamao Umezawa, /. Antibiotics 

(Japan) 8A 105 (1955). 

285 Tertiomycin B, C4;5H7iOi7N (proposed), white needles, m.p. 

97°, [a]n" -56° (c 1 in ethanol). 

A macrolide antibiotic. 

Streptomyces euroddicus 

The same organism produces eurocidin, tertiomycin A 
and azomycin. 

Teisuke Osato, Koki Yagishita and Hamao Umezawa, /. An- 
tibiotics (Japan) 8A 161 (1955). 

Akira Miyoke, Hidesuke Iwasaki and Torao Tawewaka, J. 
Antibiotics (Japan) 12A 59 (1959). 

286 Foromacidin A (Spiramycin I): C4gH780i5N2, colorless powder, 

m.p. 134-138°, [a]„ -81° (c 0.34 in methanol). 

287 Foromacidin B (Spiramycin II): C47H;^nOi6No, colorless pow- 

der, m.p. 130-132°, [a]D -83° (c 0.82 in ethanol). 

288 Foromacidin C (Spiramycin III): C4,sHs20i6N2, colorless pow- 

der, m.p. 124-128°, [a]D -79° (c 1.19 in ethanol). 

289 Foromacidin D: Equiv. Wt. 452, colorless powder, m.p. 135— 

140°, [alo -75° (c 0.81 in ethanol). 

Two streptomycetes 

R. Corbaz, L. Ettlinger, E. Gaumann, W. Keller-Schierlein, 
F. Kradolfer, E. Kyburz, L. Neipp, V. Prelog, A. Wettstein and 
H. Zahner, Helv. Chim. Acta 39 304 (1956). 

The foromacidins (or spiramycins) are apparently 
macrolide antibiotics. On degradation they yield three 
sugars typical of this class. 



Pfizer Handbook of Microbial Metabolites 



140 



Spiramycins 


[1 
II 


C«H780,5N2 
C47H80O16N2 
C48H82O16N2 


HO j"' 


Neosp 


i 
ramycins 


1: 


C38H66O12N2 
C40H68O,3N2 + 
C41H70O13N2 


OH 

CHj OH 
My ca rose 
C7HMO4 

(CHaliN 


Foro( 


:idins 


{I 


CaoHsiOuN 
C32H53O12N + 
C33H55O12N 


CH3 OH 
CsHnOzN 



Raymond Paul and Serge TchelitchefF, Bull. soc. chim. 
France 442, 734 (1957). 
Idem., ibid., 150 (1960). 

290 Tylosin, C45H-9O17N, colorless crystals, m.p. 128-132°, [ajn'" 

—46° (c 2 in methanol). 

A macrolide antibiotic, containing the sugars mycarose 
and mycaminose. Also has an a, /?, y, 8-unsaturated car- 
bonyl system. 

Streptomyces fradiae 

R. L. Hamill, M. E. Haney, Martha C. Stamper and Paul 
Wiley, Abstr. Atlantic City Meeting, Am. Chem. Soc, Septem- 
ber, 1959. (To be published) 

J. M. McGuire, W. S. Boniece, W. A. Daily, C. E. Higgens, 
M. M. Hoehn, W. M. Stark, W. B. Sutton, J. Westhead and 
R. N. Wolfe- (To be published) 

291 Angolamycin, C49_5oHs7_9iOisN, colorless crystals, m.p. 165— 

168°, [aln'^ -64° (c 1.3 in chloroform). 

A macrolide antibiotic apparently similar to carbomy- 
cin, but with characteristic sugars. 

Streptoviyces eurythermus 

R. Corbaz, L. Ettlinger, E. Gaumann, W. Keller-Schierlein, 
L. Neipp, V. Prelog, P. Reusser and H. Zahner, Helv. Chim. 
Acta 38 1202 (1955). 



292 Miamycin, colorless crystals, m.p. 221° (dec), [a]i)^^ 
1.0 in 0.02 N hydrochloric acid). 



18° (c 



^41 Other Macrolides 



A macrolide antibiotic. Analysis: C 61.4, 6i;5, H 8.7 
8.6. Mol. wt. ~609. 

Streptomyces ambofaciens 

H. Schmitz, M. Misiek, B. Heinemann, J. Lein and I. R. 
Hooper, Antibiotics and Chemotherapy 7 37 (1957). 



8 



Alicyclic Compounds Other Than 
Terpenoids and Steroids 



This section contains non-terpenoid, non-steroid alicyclics of 
diverse biosynthetic origin. Many of these, especially the strep- 
tomycete products, were antibiotic isolates. 

Included here are some of the intermediates in the biosyn- 
thetic route from carbohydrates to aromatic amino acids and to 
certain other aromatic compounds. Part of this sequence, 
worked out largely by Tatum, Davis, Sprinson and collabora- 
tors,^' -' ^ is reproduced below in brief outline only since it has 
been widely reviewed and publicized. (P indicates phosphoryla- 
tion ) : 



CHoOP 

I 

c=o 

HO— C— H 

H— C— OH 

H— C— OH 

H— C— OH 

CH2OP 



Sedoheptulose 
1, /-Diphosphate 



COOH 

I 

C— O— P 

CH. 

Phosphoenol- 
pyruvic Acid 

+ 
HC=0 

HC—OH 

HC -OH 

CH2— O— P 

Erythrose 
Phosphate 



COOH 

c=o 

CH.2 
HO— C— H 
H— C— OH 
H— C—OH 

CH2-O— P 



2-Keto-3-deoxy- 

D-araboheptonic 

Acid 



^ Bernard D. Davis, Inlermediates in ammo acid biosynthesis. Ad- 
vances in Enzymology 16 247-312 (1955). (A review) 

^ Alton Meister, "Biochemistry of the Amino Acids," Academic 
Press, Inc., New York, 1957, pp. 346-349. 

■'' P. Pi. Srinivasan, Masayuki Katagiri and David B. Sprinson, 
}. Biol. Chem. 2.'M 713 (1959); P. R. Srinivasan and David B. Sprin- 
son, ibid. 234 716 (1959). 



143 



Alicylic Compounds (Non-terpenoid) 



COOH 


COOH" 


c=o 


c=o 

1 


CH, 


1 
CH, 

1 


HO— C— H 


HO— C— H 

. 1 


c=o 

1 


-^ 1 

H— C— OH 

1 


H— C— OH 

1 


1 

c=o 

1 


CH,— O— P 


1 

CHj _ 



Aldol-like 
conden- HO 
sation 




Dehydroquinic 
Acid 

i 




Quinic Acid 



COOH 



OH 



HO 



Dehydroshikimic 
Acid 




COOH 




r\/ 



COOH 



HO 



HO 

3,4-Dihydroxy benzoic 
Acid 



Prephenic 
Acid 

i 



COOH 



r\/ 



CHoCOCOOH 



NHo 

Anthranilic 
Acid 



Phenyipyruvic 
Acid 



Phenylalanine 

Tyrosine 

Tryptophan 



Homogentisic Acid 
p-Aminobenzoic Acid 
p-Hydroxybenzoic Acid 



Microorganisms were the principal tools in this work, es- 
pecially the mold Neiirospora crassa and the bacteria Esch- 
erichia coli and Aerobacter aerogenes mutated so that the bio- 
synthesis of aromatic amino acids was blocked at various points. 
These mutants accumulated intermediates in the sequence prior 
to the blocks, and these substances could then be isolated. Also 
when such mutants (auxotrophs) were supplied with the critical 
substance whose biosynthesis was blocked, the microorganisms 
were capable of completing the sequence to the aromatic acids. 

This route from carbohydrates to certain types of aromatic 
substances has been established as quite general in metabolism. 

Biosynthesis of the chlorinated cyclopentane, caldariomycin. 



Pfizer Handbook of Microbial Metabolites 



144 



has been studied.'' ^-Ketoadipic acid and S-chlorolevulinic acid 
were found to be intermediates. The sequence shown here, 
then, probably represents at least part of the biogenetic scheme 
for this metabolite. 



COOH 

I 
OH CH2 

I I 

o=c c=o 



CHo— CH2 
/3-Ketoadipic Acid 




5-Chlorolevulinic Acid 



Caidariomycin 

Palitantin appears to be an interesting example of an un- 
aromatized acetate derivative. Its origin is revealed by the 14- 
carbon atoms, the uneven-numbered side-chains and the pattern 
of oxidation and unsaturation. 

The cycloheximides also seem to be acetate derivatives, al- 
though apparently no study of their biosynthesis has been pub- 
lished. 

Without having made a detailed analysis of the experimental 
work it would seem that the proposed structures for the glau- 
conic acids are unique if not improbable. 

293 Caidariomycin,- CgHsOoClo, colorless needles, m.p. 121°, [a]r,46i^'' 
+59.2° (c 0.338 in water). 



CI 



CI 



HCOH 

I 
-CH2 



HOCH 
CH2— 

Caldariomyces fumago 

* Paul D. Shaw, Jonathon R. Beckwith and Lowell P. Hager, /. Biol. 
Chem. 234 2560 (1959). 



145 Alicylic Compounds (Non-terpenoid) 

Percival W. Clutterbuck, Sudhir L. Mukhopadhyay, Al- 
bert E. Oxford and Harold Raistrick, Biochem. }. 34 664 
(1940). 

294 Sarkomycin, CyHsO.^, oil ( dihydro-derivative ) , m.p. 99° with sub- 
limation, [a]i."'' +66.7° (in water). 

COOH 




Streptomyces erythrochromogenes 

A yield of about 5 g. from 2 liters of broth has been 
reported. 

Hamao Umezawa, Tadashi Yamamato, Tomio Takeushi, 
Teisuke Osato, Yashiro Okami, Seizaburo Yamaoka, Tomoharu 
Okuda, Kazuo Nitta, Koki Yagishita, Ryazo Utahara and 
Sumio Umezawa, Antibiotics and Chemotherapy 4 514 (1954). 
(Isolation) 

I. R. Hooper, L. C. Cheney, M. J. Cron, O. B. Fardlg, D. A. 
Johnson, D. L. Johnson, F. M. Palermiti, H. Schmitz and 
W. B. Wheatley, ibid. 5 585 (1955). (Structure) 

M. M. Shemyakin, L. A. Shchukina, E. I. Vinogradova, 
M. N. Kolosov, R. G. Vdovina, M. G. Karapetyan, V. Ya. 
Rodionov, G. A. Ravdel, Yu. B. Shvetsov, E. M. Bamdas, E. S. 
Chaman, K. M. Ermolaev and E. P. Semkin, Zhiir. Obschchei 
Khim. 27 742 (1957). (Synthesis of dihydrosarkomycin) 

295 Terrain, CsHioOs, m.p. 127°, [a]546i'° +185° (c 1 in water). 




Aspergillus terreus Thom, Penicillium raistrickii 
Harold Raistrick and Geo. Smith, Biochem. J. 29 606 

(1935). (Isolation) 

D. H. R. Barton and E. Miller, J. Chem. Soc, 1028 (1955). 

(Structure) 



Pfizer Handbook of Microbial Metabolites 146 

296 5-Dehydroshikimic Acid, CyHj^Or,, colorless prisms, m.p. 150- 
152°, [aju'^ -57° (in ethanol). 

COOH 



HO I OH 

O 

H 

Escherichia coli mutants 

Ivan I. Salamon and Bernard D. Davis, J. Am. Chem. Soc. 
75 5567 (1953). 

297 Shikimic Acid, C7H10O5, colorless crystals, m.p. 184°, [alc^" 
-246° (in water). 



COOH 




Escherichia coli 

Yields of about 0.5 g. per liter have been reported. 
P. R. Srinivasan, Harold T. Shigeura, Milton Sprescher, 
David B. Sprinson and Bernard D. Davis, /. Biol. Chem. 220 

477 (1956). 

298 5-Dehydroquinic Acid, C^HioOg, colorless crystals, m.p. 140- 
142°. 



COOH 




Escherichia coli 

Ulrich Weiss, Bernard D. Davis and Elizabeth S. Mingioli, 
J. Am. Chem. Soc. 75 5572 (1953). 



147 Alicylic Compounds (Non-terpenoid) 

299 Dihydroshikimic Acid, C^Hi^O-,, colorless prisms, m.p. 135°, 
[a],;--' -63° (c 10 in water). 



COOH 




Lactobacillus pastoriamis var. quinicus 
A 96% yield was reported. 

J. G. Carr, A. PoUard, G. C. Whiting and A. H. Williams, 
Biochem. J. 66 283 (1957). 

300 Cordycepic Acid, CyHjoOe, colorless needles, m.p. 168°, [aln''' 
+6.8° (in water). 



COOH 




Cordyceps siriensis (Berkeley) Saccardo 
The yield was 7% of the weight of the dried and de- 
fatted mycelium. 

R. Chatterjee, K. S. Srinivasan and P. C. Maiti, /. Am. 
Pharm. Assoc. 46 114 (1957). 

301 Prephenic Acid, CiyHioOe, unstable in aqueous solution, iso- 
lated as the barium salt. 



HOOC CH2COCOOH 




Mutants of Escherichia coli and Neurospora crassa 
Ulrich Weiss, Charles Gilvarg, Elizabeth S. Mingioli and 
Bernard D. Davis, Science 119 774 (1954). 



Pfizer Handbook of Microbial Metabolites 148 

302 Frequentin, C14H20O4, colorless needles, m.p. 128°, [aln^^ +68° 

(0.5 in chloroform). 

Probably similar to palitantin in structure. 

Penicillium freqiientans Westling, P. cyclopium 

P. J. Curtis, H. G. Hemming and W. K. Smith, Nature 167 

557(1951). 

303 Palitantin, C14H22O4, colorless needles, m.p. 163°, [a]546i"^ +4.4° 

(c 0.8 in chloroform). 

O 





HOCH2 II 


OH 


CH CH r 

- \ / \ z"^-^-- 

CH CH 


OH 



CH2 

/ \ 

CH3 CH2 

Penicillium palitans Westling, P. frequentans, P. cy- 
clopium 

John Howard Birkinshaw and Harold Raistrick, Biochem. J. 
30 801 (1936). 

P. J. Curtis, H. G. Hemming and W. K. Smith, Nature 167 
557 (1951). 

A. Bracken, Anna Pocker and H. Raistrick, Biochem. J. 57 
587 (1954). 

K. Bawden, B. Lythgoe and D. J. S. Marsden, /. Chem. Soc, 
1162 (1959). (Structure) 

304 B-73, CigHieOoNo, colorless plates, m.p. 275°, [aln"' +3.43° (c 

0.4 in dimethylformamide). 

Negative ferric chloride test, non-fluorescent under U.V. 
light, soluble in aqueous sodium hydroxide. 

Streptomyces albulus 

Non-antibiotic compound isolated from a broth contain- 
ing cyclpheximide, 4-acetoxycycloheximide, C-73, and 
fungicidin. 

K. Rao, Abstracts, 134th Meeting of the American Chemical 
Society, Chicago, September 1958. 

305 C-73, C15H1-O4N, pale yellow needles, m.p. 199°, [a]i,^^ +5.06° 

(c 0.4 in dimethylformamide). 

Green ferric chloride test, bright yellow fluorescence in 
U.V. light, soluble in sodium hydroxide solution. 

Streptomyces albulus 

This antibiotically inert compound was isolated from a 
culture containing cycloheximide and stereoisomers, 4- 
acetoxycycloheximide, fungicidin, E-73 and B-73. 



149 Alicylic Compounds (Non-terpenoid) 

K. Rao, Abstracts, 134th Meeting of the American Chemical 
Society, Chicago, September 1958. 

306 Actiphenol, C15H17O4N, colorless crystals, m.p. 199°. 




An actidione-producing streptomycete (ETH 7796). 
R. J. Highet and V. Prelog, Helv. Chim. Acta 42 1523 
(1959). 

307 Inactone, C15H21O4N, colorless needles, m.p. 116°, [cxW^ —55° 
(c 2 in water). 




Streptomyces griseus 

Raymond Paul and Serge Tchelitcheff, Bull. soc. chim. 
France 1316 (1955). 

308 Cycloheximide (Actidione, Naramycin A), C15H03O4N, colorless 
crystals, m.p. 119.5-121°, [aU''' -3.4° (c 9^47 in ethanol). 

O 

CH— CH,— ( NH 

H 

o 



Streptomyces griseus, S. noursei 

Byron E. Leach, Jared H. Ford and Alma J. Whiffen, /. Am. 
Chem. Soc. 69 474 (1947). 

Jared H. Ford and Byron E. Leach, ibid. 70 1223 (1948). 




Pfizer Handbook of Microbial Metabolites 150 

Edmund C. Kornfeld, Reuben G. Jones and Thomas V. 
Parke, ibid. 71 150 (1949). (Structure) 

Tomoharu Okuda, Chem. Pharm. Bull. (Japan) 7 659 
(1959). (Stereochemistry) 

309 Cycloheximide Diasterioisomer, Ci-,H^,304N, colorless rectangu- 

lar plates, m.p. 100-105°, [a]u'' +12°. 

The crystal form differed from that of cycloheximide, 
and a mixture with authentic cycloheximide melted at 
85-95°. 

Streptomyces albidus 

Cycloheximide, 4-acetoxycycloheximide, two antibioti- 
cally inert compounds B-73 and C-73 and fungicidin were 
isolated from the same culture. 

K. Rao. Abstracts. 134th Meeting of the American Chemical 
Society, Chicago, September 1958. 

310 Naramycin B, Ci-,H^:^04N, colorless plates, m.p. 109°, [ajn^'^ 

+50.2° (c 2.0 in methanol). 









OH 


/ 


CH- 


-CH2— / NH 




\ 








Streptomyces sp. 

A stereoisomer of cycloheximide. 

Tomoharu Okuda, Makato Suzuki, Yoshiyuki Egawa and 
Kokichi Ashino, Chem. Pharm. Bull. (Japan) 6 328 (1958). 
(Isolation) 

Tomoharu Okuda, ibid. 7 659 (1959). (Stereochemistry) 

311 Streptovitacin A, Ci5H2sOr,N, colorless crystals, m.p. 156-159°. 

OH 

O 

CH3 Y CH— CH2 /^ 

o 

Streptomyces griseus 

T. E. Eble, M. E. Bergy, C. M. Large, R. R. Herr and W. G. 
Jackson, "Antibiotics Annual 1958-1959," Medical Encyclope- 
dia, Inc., New York, p. 555. (Isolation) 




151 Alicylic Compounds (Non-terpenoid) 

Ross R. Herr, /. Am. Chem. Soc. 81 2595 (1959). (Struc- 
ture) 

312 Streptovitacin B, Ci5Ho;50-,N, colorless crystals, m.p. 124-128°. 

OH 



CH3 




II 


CH- 


-CH 







\ 




/ 




Y 


NH 


y^ 


\^ 






v 


A 


HO 


CHs 












Streptomyces griseiis 

T. E. Eble, M. E. Bergy, C. M. Large, R. R. Herr and W. G. 
Jackson, "Antibiotics Annual 1958-1959," Medical Encyclope- 
dia, Inc., New York, p. 555. (Isolation) 

Ross R. Herr, /. Am. Chem. Soc. 81 2595 (1959). (Struc- 
ture) 

313 Streptovitacin C^, Ci-,H^;^05N, colorless crystals, m.p. 91-96°. 

OH 

O I o 

HO j CH— CH2 X 

CH3 ° 

Streptomyces griseiis 

Ross R. Herr, /. Am. Chem. Soc. 81 2595 (1959). (Struc- 
ture) 

314 Streptovitacin D, Ci-.H^.-^O-.N, colorless crystals, m.p. 67-69°. 

A ring-hydroxylated cycloheximide of unknown struc- 
ture. 

Streptomyces griseus 

Ross R. Herr, J. Am. Chem. Soc. 81 2595 (1959). 

315 Streptimidone, Ci,;H2304N, colorless crystals, m.p. 72°. 

O 

CH3 CH3 O OH ^ 

I I II I r< 

CHi^CH— C=CH— CH— C— CH2— CH— CHo— < N 

H 

o 

A streptomycete 



Pfizer Handbook of Microbial Metabolites 152 

Roger P. Frohardt, Henry W. Dion, Zbigniew L. Jukabow- 
ski, Albert Ryder, James C. French and Quentin R. Bartz, 
/. Am. Chem. Soc. 81 5500 (1959). 

E. E. Van Tamelen and V. Haarstad, J. Am. Chem. Soc. 82 
2974 (1960). (Revised structure) 

316 3-[2-(3,5-Dimethyl-5-acetoxy-2-oxocyclohexyl)-2-hydroxyethyl] glu- 
tarimide (4-Acetoxycycloheximide, E-73), CiyHssOgN, col- 
orless crystals, m.p. 140°, [<x]d~^ —8.8° (c 1.0 in methanol.) 




Streptomyces alhulus 

Two diastereoisomers of cycloheximide were isolated 
from the same broth. Fungicidin and two unknown com- 
pounds also were isolated. 

Koppaka V. Rao and Walter P. Cullen, J. Am. Chem. Soc. 
82 1127 (1960). (Isolation) 

Koppaka V. Rao, ibid. 82 1129 (1960). (Structure) 

317 Glauconic Acids. 

Glauconic Acid I, CigHoyO^, colorless crystals, m.p. 
202°, optically inactive. 
Proposed structure: 

CH3 r ' CH2CH2CH3 




/ — V^"^ 

o o 

and 

Glauconic Acid II, CisHooOe, colorless crystals, m.p. 
186°, optically inactive. 



Proposed structure: 

3 
\ 



CH3 ^"' CH2CH2CH3 



/^\ 



O O ? 



/ \C"^ 

o o 



153 



Alicylic Compounds (Non-terpenoid) 



Penicillium glaucum, P. purpurogenum 
Nadine Wijkman, Ann. 485 61 (1931). (Isolation) 
Kurt Kraft, ibid. 530 20 (1937). (Structure) 
Matao Takashima, Akira Kitajima and Kenichi Otsuka, 
Nippon Nogei-kagaku Kaishi 29 25 (1955). (Isolation from 
P. purpurogenum) (Chem. Abstr. 52 20379d) 

318 Fumagillin (Amebacilin, Fumidil) C26H34O7, colorless or pale 
yellow crystals, m.p. 189-194° (dec), [aln"' -26.6° (c 
0.25 in methanol). 

CH2 



O CH3 




-CH— CH2— CH=C 



CH3 



CH3 



OCH3 



O 

I 

o=c- 

Aspergillus fumigatus Fres. 

J. Landquist, /. Chem. Soc, 4237 (1956). 

J. McNally and D. Tarbell, /. Am. Chem. Soc. 80 3676 
(1958). 

D. Chapman and D. Tarbell, ibid. 80 3679 (1958). 

A. Cross and D. Tarbell, ibid. 80 3682 (1958). 

R. Carman, D. D. Chapman, N. J. McCorkindale, D. S. 
TarbeU, F. H. L. Varino, R. L. West and D. L. Wilson, /. Am. 
Chem. Soc. 81 3151 (1959). 

D. S. TarbeU, R. M. Carman, D. D. Chapman, K. R. Huff- 
man and N. J. McCorkindale, J. Am. Chem. Soc. 82 1005 
(1960). (Structure) 

T. E. Eble and F. R. Hanson, Antibiotics and Chemotherapy 
1 54 (1951). (Isolation) 



9 




Terpenoids and Steroids 



Ergosterol is the principal fungal sterol. It was named for its 
occurrence in ergot, and it has been isolated from a wide variety 
of other fungi as well as from lichens. It has been reported to 
be the only sterol in certain molds, ^ but it is often accompanied 
by related compounds. It has been identified also in algae. 
Some yeasts produce several per cent of their dry cell weight in 
ergosterol. Yeasts which produce large quantities of fat do not 
necessarily produce a higher proportion of ergosterol. 

There have been few reports of the isolation or detection of 
sterols in bacteria, and there is doubt as to whether bacteria 
produce sterols. A critical historical review of this question has 
been published.- Mevalonic acid is an acetate-replacing factor 
in lactobacilli, and a labeling study ■ with paper chromatography 
and spectral work on the labeled non-saponifiable lipides showed 
the presence of non-steroid, hydroxylated and unsaturated com- 
pounds with more than 15 carbon atoms. It may be that sim- 
pler substances of this sort replace sterols in bacteria. An 
artificial requirement for vitamin D2 can be induced in some 
bacteria. The resulting inhibition of growth can be reversed by 
vitamins D2, D., or suprasterol, but not by 7-dehydroergosterol 
nor by cholesterol.'^ 

Yeasts and higher fungi produce squalene and Co- to C;^! com- 
pounds, some of which have been shown to be precursors of 
cholesterol in mammalian metabolism. Some higher fungi and 
many lichens produce triterpenes or close derivatives. 

Since the availability of isotopes, which permit the tracing of 
small quantities of material, much of the biosynthetic route to 

Joseph V. Fiore, Arch. Biochem. 16 161 (1948). 
2 Audrey Fiertel and Harold P. Klein, J. Bacteriol. 78 738 (1959). 
•'* E. Kodicek, Abstracts of the Gordon Conference on Vitamins and 
Metabolism, 1958. 



1 55 Terpenoids and Steroids 

the principal mammalian sterol, cholesterol, has been worked 
out. Good reviews of this work are available.' Many of the 
proved intermediates in this route have been isolated from fungi, 
and evidently the biogenesis of ergosterol and the triterpenes is 
quite similar to that of cholesterol up to the later stages. ' 

The conversion of acetate to mevalonate follows the 
course : " '^'^ 

CHsCO— S— CoA + CHaCOCHjCO— S— CoA -^ 
Acetyl CoA Acetoacetyl CoA 



CH3 


CH3 "I 

1 


CoA— S— COCH2— C— CH2— COOH -^ 

1 


OHC— CHo— C—CHo— COOH 


1 

OH 


OH 


3-Methyl-3-oxyglutaryl CoA 

(Hydroxymethylglutaryl CoA, 

HMG-CoA) 


Mevoldic Acid 
CH3 




HOCHo— CH2— C— CH2— COOH 




OH 




Mevalonic Acid 



In the light of the newer knowledge concerning the role of 
malonyl CoA in fatty acid biosynthesis there may eventually be 
some minor modifications in this scheme. It should be men- 
tioned that mevalonic acid has been shown to be an irreversible 
intermediate in the biosynthesis of terpenoids.' "• ° 

Isopentenyl pyrophosphate, a further intermediate in the bio- 

* Louis F. Fieser and Mary Fleser, "Steroids," Reinhold Publishing 
Corp., New York, 1959, pp. 403-420. 

•'^ Pierre Crabbe, Record of Chemical Progress 20 189 (1959). 

''J. W. Cornforth, R. H. Cornforth, A. Pelter, M. G. Horning and 
G. Popjak, Tetrahedron 5 311 (1959). 

^^G. E. W. Wolstenholme and Maeve O'Conner (Eds.), "CIBA 
Foundation Symposium on the Biosynthesis of Terpenes and 
Sterols," Harry Rudney, The biosynthesis of P-hydroxy-fi-methyl- 
glutaryl coezyme A and its conversion to mevalonic acid. Little, 
Brown and Co., Boston, 1959, pp. 75-94. 

■ A. J. Birch, R. J. English, R. A. Massy-Westropp and Herchel 
Smith, Proc. Chem. Soc, 233 (1957). 

^Idem., J. Chem. Soc, 369 (1958). 

^J. Fishman, E. R. H. Jones, G. Lowe and M. C. Whiting, Proc. 
Chem. Soc, 127 (1959). 



Pfizer Handbook of Microbial Metabolites 



156 



synthetic process, apparently arises from phosphorylated meva- 
lonic acid by a concerted decarboxylation with elimination of 
the C-3-hydroxyl group, since it has been shown that no proto- 
nation of the carbon chain occurs during decarboxylation/° 



CH3 

HOCH2— CH2— C— CH2— COOH 

I 
OH 

Mevalonic Acid 



Mevalonic 
Acid 



Mevalonic 
Acid 



5-Monophosphate 5-Pyrophosphate 



Mevalonic 

Acid 
3-Phosphate- 
5-Pyrophosphate 



00 CH3 O ' 

T T I ^--N II ^Q 

HO— P— O— P— O— CH2— CH2— C— CH2— C— O ^ 

I 



OH OH 



OH 



HO— P— O— P— OH 

I i 

o o 

Mevalonic Acid Dipyrophosphate 



00 ^ CH3 

T T I 

HO— P— O— P— O— CH2— CH2— C=CH2 

I I 

OH OH 

Isopentenyl Pyrophosphate 

Since both y,y-dimethylallyl pyrophosphate^^ and farnesyl 
pyrophosphate^^ have been Isolated, it is possible to envisage a 
continuation : 



H3O6P2O— CH2 



H3O6P2O 



e 




CH, 



®< 



Isopentenyl 
Pyrophosphate 



N 



CH2— OP2O6H3 



\ 



Dimethylallyl 
Pyrophosphate 



Isopentenyl 
Pyrophosphate 



^° A. de Waard, A. H. Phillips and Konrad Bloch, /. Am. Chem. 
Soc. 81 2913 (1959). 

11 B. W. Agranoff, H. Eggerer, U. Henning and F. Lynen, 7- Am. 
Chem. Soc. 81 1254 (1959). 

^2 F. Lynen, H. Eggerer, U. Henning and Ingrid Kessel, Angew. 
Chem. 70 738 (1958). 



157 



Terpenoids and Steroids 



CHj— OP2O6H3" 




CH2— OP2O6H3 




Farnesyl Pyrophosphate 

Two moles of farnesyl pyrophosphate then unite head-to-head 
in what, deuterium experiments indicate/^- " is probably a re- 
ductive process to form squalene.* All trans-squalene is formed, 
and this is the only isomer which can cyclize to triterpenes and 
steroids. ^^ 

The significance of the stereoisomer has been considered, and 
a generalized scheme devised for the various modes of cycliza- 
tion of squalene, supported by the current theories of conforma- 
tional analysis and ionic cyclization.^*^' "• ^® 

Squalene can cyclize with no skeletal rearrangement to form 
compounds such as the lichen substance, zeorin. It also can 
rearrange to the lanostane skeleton found so frequently among 
the steroids of the higher fungi. Lanosterol itself, a known 
intermediate in the biosynthetic route to cholesterol, has been 
found in yeast, as has squalene. 




13 H. Rilling, T. T. Tchen and Konrad Bloch, Proc. Nat. Acad. Sci. 
44 163 (1958). 

'* H. C. Rilling and Konrad Bloch, }. Biol. Chem. 234 1424 (1959). 

* See addendum for a recent modification of this scheme. 

15 Robert G. Langdon and Konrad Bloch, ibid. 200 135 (1953). 

^^ L. Ruzicka, A. Eschenmoser and H. Heusser, Experientia 9 362 
(1953). 

1^ A. Eschenmoser, L. Ruzicka, O. Jeger and D. Arigoni, Helv. 
Chim. Acta 38 1890 (1955). 

1® Alexander Todd, "Perspectives in Organic Chemistry," L. Ru- 
zicka, Bedeutung der theoretischen organischen Chemie fi'ir die 
Chemie der Terpenverbindungen, Interscience Publishers, Inc., New 
York, 1956, pp. 265-315; L. Ruzicka, Proc. Chem. Soc, 341-360 
(1959); Faraday Lecture, History of the isoprene rule. 



Pfizer Handbook of Microbial Metabolites 



158 



It is likely that the cyclization of squalene to form such struc- 
tures occurs by a concerted mechanism which proceeds from 
ring to ring until complete and that this all occurs on one 
enzyme surface. Thus, isolation of intermediates between 
squalene and an initial cyclization product such as the one 
shown is improbable. The cyclization is oxygen-initiated, ex- 
plaining the frequent occurrence of the 3-hydroxyl groups in 
natural steroids. 




HO / \ HO 

Proposed initial cyclization 

product of the Squalene —>■ Lanosterol 

route 




Lanosterol 



Transformation of the proposed tetracyclic precursor to lano- 
sterol involves two 1,2-methyl group migrations (14-^ 13 and 
8 ^ 14) as shown by tracer experiments." ^^ 

Eburicoic acid has the lanostane skeleton, but with a methyl- 
ene group attached to carbon atom 24 of the side-chain. Simi- 
larly, ergosterol has a methyl group in this position. Labeling 




CHo 



HOOC 




Ergosterol 



CH3 CH3 

Eburicoic Acid 



^'■' R. K. Maudgal, T. T. Tchen and Konrad Bloch, J. Am. Chem. 
Soc. 80 2589 (1958). 



159 



Terpenoids and Steroids 



experiments-'- -' -- ~-^ have shown that this "extra" carbon atom 
is not derived from acetate, but is furnished by formate and, 
more efficiently, by methionine. 

Progressing along the biosynthetic route from squalene to 
ergosterol (and cholesterol), it is obvious that lanosterol must 
lose the two methyl groups at C-4 and one at C-14. These are 
probably removed oxidatively, and eventually some of the inter- 
mediates may be isolated. Zymosterol has been considered as 
an intermediate in the biosynthesis of cholesterol; but while it 
occurs together with ergosterol in yeasts, it has been found-** that 
squalene, but not zymosterol, is converted to ergosterol by yeast 
homogenates. 

The biogenesis of the interesting diterpenoids gibberellic acid, 
rosenonolactone and trichothecin has been studied. In the case 
of gibbereUic acid-* studies with CH:^C"OOH and with C-2-la- 



CH3COOH 





beled m-evalonate gave the labeling pattern shown. A precursor 
was inferred, and the followdng deductions made: (a) The 
methyl carbon atom attached to ring A is derived specifically 
from position 2 of mevalonic acid lactone, (b) The carboxyl 
carbon atom is derived specifically from position 9 of the pre- 
cursor, (c) The phyllocladene ring system of gibberellic acid 
is formed by migration of C-6 to C-18. 

Rosenonolactone, rosololactone and trichothecin are all pro- 
duced by the fungus Trichothecium roseum. 

2" George J. Alexander, Allen M. Gold and Erwin Schwenk, ibid. 79 
2967 (1957). 

21 William G. Dauben and John H. Richards, ibid. 78 5329 (1956). 

2- William G. Dauben, Yoshio Ban and John H. Richards, ibid. 79 
968 (1957). 

2^ William G. Dauben, Gerhard J. Fonken and George A. Boswell, 
ibid. 79 1000 (1957). 

2* A. J. Birch, R. W. Rickards and Herchel Smith, Proc. Chem. 
Soc, 192 (1958). 



Pfizer Handbook of Microbial Metabolites 



1 60 



19 17 





COOH 




O^ 



o 

CH3— CH=CH— coo 



Rosenonolactone 



Rosololactone 



Trichothedn 



The carbon skeleton of rosenonolactone'' is apparently derived 
from the same kind of precursor as gibberellic acid, but in a 
simpler way. All that is required is the migration of a methyl 
group from C-12 to C-13 in the same manner as in the biosyn- 
thesis of steroids. 

The carbon skeleton of trichothecin^" can be derived from a 
sesquiterpenoid intermediate by way of two 1,2-methyl group 
shifts : 

-O 





2-C'^-Me- 
valonate 



Proposed sesquiter- 
penoid intermediate 



CH— COO 

Trichothecin 



319 



When labeled acetate was used in this study, 95% of the 
activity appeared in the crotonic acid moiety. These results, 
considered together, are another confirmation of the irreversibil- 
ity of the acetate-mevalonate process. 

A symposium has been published thoroughly reviewing cur- 
rent research on the biosynthesis of terpenes and sterols.^^ 
Lactaroviolin, C15H14O, red-violet crystals, m.p. 53°. 




OHC 



-=G. E. W. Wolstenholme and Maeve O'Conner (Eds.), "CIBA 
Foundation Symposium on the Biosynthesis of Terpenes and Sterols," 
Little, Brown and Co., Boston, 1959. 



i6i Terpenoids and Steroids 

Lactaiius deliciosiis L. 

Harry Willstaedt and B. Zetterberg, Svensk Kem. Tidskr. 58 
306 (1946). 

PI. A. Plattner, E. Heilbronner, R. W. Schmid. R. Sandrin 
and A. Furst, Chem. and Ind., 1202 (1954). (Structure) 

E. Heilbronner and R. W. Schmid, Helv. Chim. Acta 37 
2018 (1954). 

320 Lactarazulene, C15H16, blue liquid, b.p. 155-160° (2.5-3 mm.). 




Lactariiis deliciosiis L. 

Occurs together with iactaroviolin (q.v. ) and a green 
321 crystalline compound, Verdazulene, C15H16, m.p. 90°. 

Frantisek Sorm, Vera Benesova and Vlastimil Herout, Chem. 
Listij 47 1856 (1953). (Structure) 

Gibberellins and Gibberellenic Acid 

Although gibberellic acid is the gibberellin produced in 
highest yield by Gibberella fujikuroi, three minor gibber- 
eUins are produced also, and the crude mixture is com- 
monly isolated. The minor gibberellins are called A■^, A2 
and A4, gibberelhc acid being A3. (It also has been called 
gibberellin X.) Their structures are similar to those of 
gibberellic acid. 

Gibberellin A^ has been found in plants as well as in 
fungi. All of the four gibberellins show plant hormone 
activity. A fifth, inactive, compound called gibberellenic 
acid has been isolated recently. It may be an artifact. 

A structure for gibberellic acid was advanced in 1956 
by an English group. Structure work has continued in 
Japan, where the gibberellins were originally isolated, and 
recently structures for all the gibberellins have been pub- 
Ushed which differ somewhat from the one first advanced 
in England. Even more recently a third set of structures, 
complete with stereochemistry, has been proposed by the 
English school. It is these structures which are shown 
here. 



Pfizer Handbook of Microbial Metabolites 162 

322 Gibberellenic Acid, CjoHo^Op,, colorless crystals, m.p. 185° (dec). 
Strong U.V. absorption at 253 lU/x (e = 19,200). 





COOH 



Fusarium moniliforme 

Koert Gerzon, Harold L. Bird, Jr. and Don O. Woolf, Jr., 
Experientia 13 487 (1957). 

323 Gibberellic Acid (Gibberellln A;., Gibberellln X), C19H22O6, col- 
orless crystals, m.p. 235° (dec), [a]r,-" +92°. 
Proposed complete stereochemical structure: 



HO 



P. J. Curtis and B. E. Cross, Chem. and Ind., 1066 (1954). 
(Isolation) 

B. E. Cross, John Frederick Grove, J. MacMillan and T. P. C. 
Mulholland, Chem. and Ind., 954 (1956). (Structure) 

Brian E. Cross, /. Chem. Soc, 4670 (1954). 

Nobutaka Takahashi, Yasuo Seta, Hiroshi Kitamura and 
Yusuke Sumiki, Bull. Agr. Chem. Soc. (Japan) 23 405 (1959). 

Hiroshi Kitamura, Yasuo Seta, Nobutaka Takahashi, Akira 
Kawarada_and Yusuke Sumiki, ibid. 23 408 (1959). 

Yasuo Seta, Nobutaka Takahashi, Akira Kawarada, Hiroshi 
Kitamura and Yusuke Sumiki, ibid. 23 412 (1959). 

Nobutaka Takahashi, Yasuo Seta, Hiroshi Kitamura, Akira 
Kawarada and Yusuke Sumiki, ibid. 23 493 (1959). 

Yasuo Seta, Nobutaka Takahashi, Hiroshi Kitamura, Ma- 
koto Takai, Sahuro Tamura and Yusuke Sumiki, ibid. 23 499 
(1959). 

Nobutaka Takahashi, Yasuo Seta, Hiroshi Kitamura and 
Yusuke Sumiki, ibid. 23 509 (1959). 

B. E. Cross, J. F. Grove, P. McCloskey and T. P. C. Mulhol- 
land, Chem. and Ind., 1345 (1959); B. E. Cross, John Fred- 
erick Grove, J. MacMillan, J. S. Moffatt, T. P. C. Mulholland 
and J. C. Seaton, Proc. Chem. Soc, 302 (1959). 



1 63 



Terpenoids and Steroids 



324 Gibberellin A4, C19H04O-,, colorless crystals, m.p. 222° (dec), 
[a]u'° -20.8° (c 0.34 in methanol). 




CH3 COOH \ 

Gibberella fujikuroi (Saw.) Wollenweber 

Nobutaka Takahashi, Yasuo Seta, Hiroshi Kitamura and 

Yusuke Sumiki, Bull. Agr. Chem. Soc. (Japan) 21 396 

(1957). (Isolation of Gibberellin A,) 

See other references under Gibberellin Ai for structure 

work. 

325 Gibberellin A^ C19H24O6, colorless crystals, m.p. 255-258° 
(dec), [a]D-'+36°. 




CH3 COOH \ 



CH2 



Gibberella fujikuroi (Saw.) Wollenweber 

Nobutaka Takahashi, Hiroshi Kitamura, Akira Kawarada, 
Yasuo Seta, Makato Takai, Suburo Tamura and Yusuke Su- 
miki, Bull. Agr. Chem. Soc. (Japan) 19 267 (1955). (Isola- 
tion of gibberellins and their properties) 

Nobutaka Takahashi, Yasuo Seta, Hiroshi Kitamura and 
Yusuke Sumiki, ibid. 23 405 (1959). 

Hiroshi Kitamura, Yasuo Seta, Nobutaka Takahashi, Akira 
Kawarada and Yusuke Sumiki, ibid. 23 408 (1959). (Struc- 
tures of the gibberellins) 

Nobutaka Takahashi, Yasuo Seta, Hiroshi Kitamura, Akira 
Kawarada and Yusuke Sumiki, ibid. 23 493 (1959). (Struc- 
tures of the gibberellins). 

Yasuo Seta, Nobutaka Takahashi, Hiroshi Kitamura, Ma- 
kato Tokai, Saburo Tamura and Yusuke Sumiki, ibid. 23 499 
(1959). (Structures of the gibberellins) 

Nobutaka Takahashi, Yasuo Seta, Hiroshi Kitamura and 
Yusuke Sumiki, ibid. 23 509 (1959). (Structures of the 
gibberellins ) 



Pfizer Handbook o£ Microbial Metabolites 



164 



B. E. Cross, John Frederick Grove, J. MacMillan, J. S. Mof- 
fatt, T. P. C. Mulholland and J. C. Seaton, Proc. Chem. Soc, 
302(1959). (Above structure) 

326 Gibberellin Ao, CisHoeOg, colorless crystals, m.p. 235-237° 

(dec), [a]D +11.7°. 




Gibberella fujikuroi (Saw.) Wollenw^eber 
See references under Gibberellin Ai. 

327 Trichothecin, C19H24O5, colorless needles, m.p. 118°, [cjId^ 
+44° (c 1.0 in chloroform). 

CH2-^ 



X 



CH3 



o— c— c=c 

II H H 

o 



Trichotheciiim roseum (Link) 

G. G. Freeman and R. I. Morrison, Nature 162 30 (1948). 

G. G. Freeman, J. E. Gill and W. S. Waring, J. Chem. Soc, 
1105 (1959). (Structure) 

J. Fishman, E. R. H. Jones, G. Lowe and M. C. Whiting, 
Proc. Chem. Soc, 127 (1959). (Structure) 

328 Rosenonolactone, C00H28O3, white prisms, m.p. 208°, [aln^^ 
— 116° (c 2.0 in chloroform). 




Trichotheciiim roseum (Link) 

About 6 g. of dry mycelium were obtained from a Uter 
of culture solution, and from this about 0.2 g. of rosenono- 
lactone was extracted. 

Alexander Robertson, W. R. Smithies and Eric Tittensor, /. 
Chem. Soc, 879 (1949). (Isolation) 



165 Terpenoids and Steroids 

Adelaide Harris, Alexander Robertson and W. B. Whalley, 
ibid., 1799 (1958). (Structure) 

329 9-Deoxorosenonolactone, CooH;io02, colorless crystals, m.p. 115°, 
[a]D +57° (in chloroform). 




Trichothecium roseum (Link) 

W. B. Whalley, B. Green, D. Arigoni, J. J. Britt and Carl 
Djerassi, /. Am. Chem. Soc. 81 5520 (1959). 

330 Rosololactone, C00H30O3, white crystals, m.p. 186°, [ajn^^ +6.3° 
(c 2.3 in chloroform). 



COOH 

Trichothecium roseum (Link) 

Rosololactone is a minor product of this fermentation. 
It occurs in the mycelium together with rosenonolactone 
and mannitol. 

Alexander Robertson, W. R. Smithies and Eric Tittensor, J. 
Chem. Soc, 879 (1949). (Isolation) 

Adelaide Harris, Alexander Robertson and W. B. Whalley, 
ibid., 1807 (1958). (Structure) 

331 Zymosterol, C27H44O, colorless crystals, m.p. 110°, [ajo +49°. 



HO 

Zymosterol is second to ergosterol in abundance in 
yeast fat. 



Pfizer Handbook of Microbial Metabolites i66 

Ida Smedley-MacLean, Biochem. J. 22 22 (1928). (Isola- 
tion) 

332 Hyposterol, tentatively C27H40O or C27H44O, colorless unstable 
crystals, m.p. 100-102°, [y.]u-° +12.5° (in chloroform). 
Structure unknown. May be a C^s sterol. 
Yeasts 
Heinrich Wieland and G. A. C. Gaugh, Ann. 482 36 (1930). 

133 Anasterol, C27H44O, colorless crystals, m.p. 157-159°, [a]D^^ 
—8.1° (in chloroform). 

Structure unknown. May be a C2S sterol. 

Yeasts 

Heinrich Wieland and G. A. C. Gaugh, Ann. 482 36 (1930). 

334 14-Dehydroergosterol, C08H42O, colorless needles, m.p. 198- 
201°, [a]D -396° (in carbon tetrachloride). 



^ 



/ 
HO 

Aspergillus niger 

Ergosterol was isolated from the same culture. 

D. H. R. Barton and T. Bruun, J. Chem. Soc, 2728 (1951). 

335 24(28)-Dehydroergosterol (5,7,22,24(28)-Ergostatetraen-3-^-ol), 
C2SH42O, colorless crystals (Monohydrate), m.p. 118-120°, 
[aln'' -78° (1% in chloroform). 
Probable -structure : 



HO 

Yeasts 

Under appropriate growth conditions, yields of this 
sterol equal those of ergosterol. 



1 67 Terpenoids and Steroids 

O. N. Breivek, J. L. Owades and R. F. Light, J. Org. Chem. 
19 1734 (1954). 

336 Ergosterol, C0SH44O, colorless crystals, m.p. 165°, [a],^^ —130° 
(in chloroform). 



HO 

Ergosterol is the most abundant sterol of yeasts and 
molds. It occurs widely and was isolated first from ergot 
(Claviceps purpurea) . It also occurs in lichens and has 
been detected in Euglena spp. There is much literature, 
one recent example being: 

Akira Saito, /. Fermentation Technol. (Japan) 31 140 
(1953). 

Yields as high as 2-2.7% of dry cell weight have been 
reported, by using Saccharomyces carlsbergensis. Ergos- 
terol is reported to be the only sterol occurring in several 
species of Fusaria. It occurs as the palmitate in Peni- 
cillium spp. and in Aspergillus fumigatus. 

Albert E. Oxford and Harold Raistrick, Biochem. J. 27 1176 
(1933). 

P. Wieland and V. Prelog, Helv. Chim. Acta 30 1028 (1947). 

Joseph V. Flore, Arch. Biochem. 16 161 (1948). 

337 Pyrocalciferol, C2SH44O, colorless needles, m.p. 93-95°, [alo^" 
+502° (in alcohol). 




HO 

Penicillium notatum 

A yield of 12 mg. was obtained from 450 g. of dry 
mycelium. 



Pfizer Handbook of Microbial Metabolites 



1 68 



A. Angeletti, G. Tappi and G. Biglino, Ann. Chim. (Rome) 
42 502 (1952). 

J. CasteUs, E. R. H. Jones, G. D. Meakins and R. W. J. Wil- 
liams, J. Chem. Soc, 1159 (1959). (Structure) 

338 Ergosta-7,22-dien-3-one, C08H44O, m.p. 184-187°, [a]u +6° (in 
chloroform ) . 



/^ 




/ 



Fomes fomentarius 

H. R. Arthur, T. G. Halsall and R. D. Smith, /. Chem. Soc, 
2603 (1958). 

339 Ergosterol Peroxide, C28H44O3, colorless crystals, m.p. 178°, [ajn 
-36°. 




Aspergillus fumigatus (mycelium) 

P. Wieland and V. Prelog, Helv. Chim. Acta 30 1028 (1947). 

340 Episterol (A''^**"^''-Ergostadien-3/3-ol), CosH4qO, colorless crystals, 
m.p. 150°, [(x]d —5° (in chloroform). 



HO 



Yeasts 



1 69 



Terpenoids and Steroids 



Heinrich Wieland, Fridolf Rath and Horst Hesse, Ann. 548 
34 (1941). 

341 5,6-Dihydroergosterol (A'"--Ergostadien-3/?-ol), C2sH4(;0, color- 
less crystals, m.p. 176°, [a]i)-° '^ —19° (in chloroform). 




HO 

Yeasts, Claviceps purpurea 

Heinrich Wieland and Willi Benend, Ann. 554 1 (1943). 

D. H. R. Barton and J. D. Cox, /. Chem. Soc, 1354 (1948). 

342 Fecosterol (A^'^**^^*'-Ergostadien-3^-ol), C28H46O, colorless crys- 
tals, m.p. 161-163°, [alo'' -f 42° (in chloroform). 



C^ 



HO 
Yeasts 

Heinrich Wieland, Fridolf Rath and Horst Hesse, Ann. 548 
34 (1941). 

D. H. R. Barton and J. D. Cox, /. Chem. Soc, 214 (1949). 

343 Ascosterol (A^^^"'-Ergostadien-3^-ol), C28H46O, colorless crystals, 
m.p. 146°, [aW^ +45° (in chloro'form). 



HO 



Yeasts 



Pfizer Handbook of Microbial Metabolites 



170 



Heinrich Wieland, Fridolf Rath and Horst Hesse, Ann. 548 
34 (1951). 

344 Cerevisterol (A' ---Ergostadiene-3;3,5a,6/3-triol), Co,sH4,.03, color- 
less crystals, m.p. 256-259°, [a]n -83° (c 0.89 in pyri- 
dine). 




H OH 



Yeasts, Claviceps purpurea (ergot), Amanita phalloides 

About 10 g. were obtained from 4500 kg. of dried yeast. 
Some 20 g. were obtained from 17 kg. of dry Amanita 
phalloides. 

Heinrich Wieland and Gustav Coutelle, Ann. 548 275 
(1941). (Isolation) 

G. H. Alt and D. H. R. Barton, /. Chem. Soc, 1356 (1954). 
(Structure) 

345 Fungisterol (A'-Ergosten-3^-ol), C^sH4sO, colorless crystals, m.p. 
149°, [a]i,23 -0.2° (in chloroform). 



HO 

Fungisterol accompanies ergosterol in ergot, occurs in 
Amanita phalloides, Penicillium. chrysogenum, Rhizopus 
saponicus, Calocera viscosa, Polyporus confiuens Ft., P. 
sulfureus (Bull.) Fr., Hydnum imbricatum L., Geaster 
fimbriatus Fr. 

Heinrich Wieland and Gustav Coutelle, Anii. 548 270 
(1941). (Structure) 

Akira Saito, J. Fermentation Technol. (Japan) 29 310 
(1951). 



lyi 



Terpenoids and Steroids 



346 21 -Hydroxy lanosta-7,9(ll)-24-triene-3-one, CioH^e.Oa, colorless 
needles, m.p. 119-121°, [alo +56° (c 0.97 in chloroform). 



HOCH 




Polyporus pinicola Fr. 

The derivative reduced and acetylated in the 3-position 
was isolated from the same specimen as were fungisterol 
and ergosta-7,22-diene-3-one. 

T. G. Halsall and G. C. Sayer, }. Chem. Soc, 2031 (1959). 

347 Pinicolic Acid A, C3oH4(.03, colorless needles, m.p. 197-202°, 
[a]D +68° (c 0.83 in chloroform). 



HOOC 




Polyporus pinicola Fr. 

Joyce M. Guider, T. G. Halsall and E. R. H. Jones, /. Chem. 
Soc, 4471 (1954). 

348 Lanosta-7,9(ll)-24-triene-3^,21-diol, C3„H4sOo, colorless needles, 
m.p. 194-197°, [x]d +72° (c 1.06 in chloroform). 



HOCH, 




Pfizer Handbook of Microbial Metabolites 



172 



Polyporus pinicola Fr. 

The corresponding 3-ketone was isolated from the same 
specimen as well as a mixture of fungisterol and ergosta- 
7,22-diene-3-one. 

T. G. Halsall and G. C. Sayer, /. Chem. Soc, 2031 (1959). 

349 3/3-Hydroxylanosta-8,24-diene-21-oic Acid (Trametenolic Acid 
B), C30H48O3, colorless needles, m.p. 253-258°, [aU +43° 



(c 0.94 in pyridine). 



HOOC 




Trametes odorata (Wulf. ) Fr. 

Three other acids were isolated as their methyl esters 
from the same specimen: Ester A; m.p. 159-165°, [ci:]d 
+49°. Ester B, m.p. 152°, [ajn +66° and Ester C, m.p. 
197-199°. 

T. G. Halsall, R. Hodges and G. C. Sayer, J. Chem. Soc, 
2036 (1959). 

T. G. Halsall and E. R. H. Jones, Fortschr. Chem. org. 
Naturst. 12 95 (1955). (A review) 

350 3a-Oxylanosta-8,24-diene-21-oic Acid, C30H4SO3, isolated as the 
methyl ester-acetate. 



HOOC 




Polyporus pinicola Fr. 

J. J. Beereboom, H. Fazakerley and T. G. Halsall, J. Chem. 
Soc, 3437 (1957). 



1 73 Terpenoids and Steroids 

351 Squalene, C^oH-.o, pale yellow oil with blue fluorescence, b.p. 
203° (0.15 mm.), Hd^" 1.4965. Often isolated as the hy- 
drogen chloride addition product. 




Yeasts, Claviceps purpurea (ergot), Amanita phalloides 

Squalene may constitute as much as 16% of brewers' 
yeast lipide. 

A. H. Cook, "The Chemistry and Biology of Yeasts," A. A. 
Eddy, Aspects of the chemical composition of yeast. Academic 
Press, New York, 1958, pp. 207-208. 

K. Taufel, H. Thaler and H. Schreyegg, Fettchem. Umschau 
43 26 (1936). 

About 3 g. were obtained from 17 kg. of Amanita 
phalloides. 

Heinrich Wieland and Gustav Coutelle, Ann. 548 275 
(1941). 

Nearly 25% of the unsaponifiable fraction of the fat of 
Torula utilis were found to be squalene. 

R. Reichert, Helv. Chim. Acta 28 484 (1945). 

352 Lanosterol (Kryptosterol, A®'^*-Lanostadien-3-ol), C30H50O, color- 
less crystals, m.p. 138°, [ajn +62° (in chloroform). 




Yeasts 

Heinrich Wieland, Heinrich Pasedach and Albert Ballauf, 
Ann. 529 68 (1937). 



Pfizer Handbook of Microbial Metabolites 



174 



L. Ruzicka, R. Denss and O. Jeger, Helv. Chim. Acta 29 204 
(1946). 

W. Voser, M. V. Mijovic, H. Heusser, O. Jeger and L. Ru- 
zicka, ibid. 35 2414 (1952). 

353 Physarosterol, C3i,H-,.0;i, colorless crystals, m.p. 137-139°, [cxW^ 

—55.3° (c 0.5 in chloroform). 

Probably a C3,,, unsaturated, trihydroxy sterol with one 
hydroxyl group in the 3y3-position. 

Pliysarum polycephaliim 

This organism also produces a yellow pigment. 

C. F. Emanuel, Nature 182 1234 (1958). 

354 Polyporenic Acid C, C;^iH4(j04, colorless crystals, m.p. 273-276°, 

[a]D +8° (in pyridine). 



HOOC 




Polyporus betulirms, P. benzoinus, P. pinicola 
A. Bowers, T. G. Halsall and G. C. Sayer, /. Chem. Soc, 
3070 (1954). 

355 Agaricolic Acid, C;^,H4sO;^ (probably), colorless crystals, m.p. 

226°, [or],,-" +34.4° (in pyridine). 

Probably a monohydroxy triterpene acid. It occurs 
together with ergosterol and eburicoic acid, agaricic acid 
and other metabolites. 

Polyporus officinalis 

J. Valentin and S. Kniilter, Pharm. Zentralhalle 96 478 
(1957). 

356 Dehydrotumulosic Acid, C31H4SO4. 

HOOC 




175 



Terpenoids and Steroids 



Polyporus tumulosus Cooke, P. australiensis Wakefield, 
P. betuliuus. Porta cocos 

This acid has never been separated completely from its 
mixture with tumulosic aicd, but the structure has been 
deduced from physical measurements. 

L. A. Cort, R. M. Gascoigne, J. S. E. Holker, B. J. Ralph, 
Alexander Robertson and J. J. H. Simes, J. Chem. Soc, 3713 
(1954). 

357 Eburicoic Acid, C;,,H-,„0;., colorless crystals, m.p. 292-293°, 
[a]i,'" +50° (c 1.2 in chloroform). 



HOOC 




Polyporus officinalis Fr., P. anthracophihis, Cooke, P. 
eucalyptorum Fr., P. sulfiireiis (Bull.) Fr., P. hispidus 
(Bull.) Fr., Porta cocos (Schw.) Wolf, Lentinus dacty- 
loides Cleland. 

The yield is 50% of the dry mycelial weight in some 
cases. The S^g-acetate also occurs naturally in at least 
some of these basidiomycetes. 

J. S. E. Holker, A. D. G. Powell, Alexander Robertson, 
J. J. H. Simes, R. S. Wright and (in part) R. M. Gascoigne, /. 
Chem. Soc, 2422 (1953). (Structure) 

358 Tumulosic Acid, Ci^H-.^O^, colorless fine needles, m.p. 306° 
(dec), [a]„ +8.1° (c 3.30 in pyridine). 



HOOC 




Polyporus tumulosus Cooke, P. australiensis Wakefield, 
P. betulinus Fr., Porta cocos Wolf, Porta cocos (Schw.) 
Wolf (syn. Pachyma hoelen Rumph.) 



Pfizer Handbook of Microbial Metabolites 



176 



L. A. Cort, R. M. Gascoigne, J. S. E. Holker, B. J. Ralph, 
Alexander Robertson and J. J. H. Simes, /. Chem. Soc, 3713 

(1954). 

359 Polyporenic Acid A (Ungulinic Acid), C31H50O4, colorless nee- 
dles, m.p. 199-200° (dec), [o^W +64° (c 1.28 in pyri- 
dine). 

COOH 
HO ^ 




HO 

Polyporus betulinus Fr. 

T. G. HalsaU and R. Hodge, /. Chem. Soc, 2385 (1954). 
( Structure ) 

360 O-Acetyleburicoic Acid, C33H50O4, colorless needles, m.p. 256— 
259°, [aln'' +33.4° (in pyridine). 

HOOC 




CH3— C— O 

Polyporus anthracophilus 

R. M. Gascoigne, J. S. E. Holker, B. J. Ralph and Alexander 
Robertson,-/. Chem. Soc, 2346 (1951). 

F. N. Lakey and P. H. A. Strasser, ibid., 873 (1951). 
(Structure) 

361 Ursolic Acid (probable structure), C3(jH4).03, colorless crystals, 
m.p. 291-292°, [(xU~' +72° (in 1:1 chloroform-methanol). 




177 



Terpenoids and Steroids 



Cladonia sylvatica L. Harm., CI. impexa Harm. 

This acid also occurs in animals and plants. Since 
pentacyclic triterpenes are not characteristic of molds, 
they may be produced by the algal partner of the symbiont 
lichen. 

T. W. Breaden, J. Keane and T. J. Nolan, Sci. Proc. Roy. 
Dublin Soc. 23 197 (1944). 

A. Ziircher, O. Jerger and L. Ruzicka, Helv. Chim. Acta 37 
2145 (1954). 

362 Taraxerene, CgoHjo, colorless crystals, m.p. 237°, [ajo +1° (c 
0.81 in chloroform). 




Cladonia deformis Hoflm. 

About 15 mg. of pure hydrocarbon were obtained from 
2.9 kg. of dry lichen. 

Torger Bruun, Acta Chem. Scand. 8 71 (1954). 



363 Friedelin, C30H50O, colorless crystals, m.p. 267-269' 
(vac), [a]D -21° (c 2.34 in chloroform). 



(dec.) 




^Xp^ 



Cetraria nivalis (L.) Ach., C. islandica (L. ) Ach., C. 
cucullata (Bell.) Ach., C. crispa (Ach.) Nyl., C. delisei 
(Bory) Th. Fr. (syn. hiascens Fr. ), Cladonia alpestris (L.) 
Rabh., Alectoria ochroleuca (Ehrh.) Nyl. and Stereocaulon 
paschale (L.) Fr. 

Torger Bruun, Acta Chem. Scand. 8 71 (1954). 



Pfizer Handbook of Microbial Metabolites 



178 



364 epi-Friedelinol, CgoH-.oO, colorless crystals, m.p. 280° (vac), 
[a]D +23° (c 0.52 in chloroform). 




Cetraria nivalis (L.) Ach. 

Torger Bruun, Acta Chem. Scand. 8 71 (1954). 

365 Zeorin, C:hoH-.202, colorless crystals, m.p. 223-227°, [ajo +54^ 
(c 0.50 in chloroform). 




366 



Lecanora sordida, L. thiodes, L. epanora, L. sulfiirea, 
Physcia caesia. Ph. endococcina, Anaptychia speciosa, A. 
hypoleuca, Parmelia leiicotyliza, Dimelaena oreina, 
Haematomma coccineum, H. leiphaemum, H. prophyrium, 
Placodium saxicolum, Peltigera malacea, P. horizontalis , 
P. propagiilifera. Nephroma arcticum, N. antarcticum, 
N. laevigatum, N. parile, Cladonia deformis, Coccifera 
pleurota, C. helUdiflora, Urceolaria cretacea, Lepraria 
latebrarum 

"Elsevier's Encyclopedia of Organic Chemistry," 14 Suppl., 
Elsevier Publishing Co., London, 1952, p. 1197S. (Occur- 
rence) 

D. H. R. Barton and T. Bruun, /. Chem. Soc, 1683 (1952). 

D. H. R. Barton, P. de Mayo and J. C. Orr, ibid., 2239 
(1958). 

Lcucotylin, C:^„H-,20:^, colorless prisms, m.p. 333°, [a]i.-* +49.43°, 
A triterpenoid compound accompanying zeorin. 
Parvielia leiicotyliza Nyl. 
Yasuhiko Asahina and Hirosi Akagi, Ber. 71B 980 (1938). 



J 79 



Terpenoids and Steroids 



367 Helvolic Acid (Fumigacin, May = Mycocidin), C;{2H420s, color- 
less fine needles, m.p. 211° (dec). [oc]u-'' -124° (in chlo- 
roform ) . 

Tentative partial structure: 



HOOC 



^— CHo— 




Aspergillus fuinigatus mut. helvola Yuill 

Donald J. Cram and Norman L. Allinger, /. Am. Chem. Soc. 
78 5275 (1956). (Structure) 

E. Chain, H. W. Florey, M. A. Jennings and T. 1. Williams, 
Brit. ]. Exp. Pathol. 24 108 (1943). (Isolation) 

CIBA Lectures in Microbial Chemistry, E. P. Abraham, 
"Biochemistry of Some Peptide and Steroid Antibiotics," The 
cephalosporins, John Wiley and Sons, New York, 1957, pp. 
30-63. (A review) 

Cephalosporins P 

These non-peptide compounds accompany cephalospo- 
rins N and C in Cephalosporium salmosynnematum fer- 
mentations. 



TABLE I 



368 



369 
370 



371 



Compound 


Crystal 
form 


Melting 
point 


Wn 


Molecular 
formula 


Cephalosporin Pi 


colorless nee- 
dles 


147° 


+ 28° (c 2.7 in 
chloroform) 


C32H48O8 


Cephalosporin P2 




151° 






Cephalosporin P3 


white, amor- 
phous solid 








Cephalosporin P4 


Fawn-colored 
crystals 


220-230° 







Cephalosporin Pj resembles helvolic acid (from Asper- 
gillus fuinigatus). A complete (steroid) structure has 
been determined by T. G. Halsall and associates, but has 
not been published yet. 



Pfizer Handbook of Microbial Metabolites i8o 

H. S. Burton and E. P. Abraham, Biochem. J. 50 168 
(1951). (Isolation) 

H. S. Burton, E. P. Abraham and H. M. E. Cardwell, ibid. 
62 171 (1956). 

CIBA Lectures in Microbial Biochemistry, E. P. Abraham, 
"Biochemistry of Some Peptide and Steroid Antibiotics," The 
cephalosporins, John Wiley and Sons, New York, 1957, pp. 
30-63. (A review) 

368 Cephalosporin Pj, C32H480,s, colorless crystals, m.p. 147°, [aln^" 
+28° (2.7 in chloroform). 

Cephalosporium spp. 

A number of similar substances, called cephalosporins 
P2, P3, P4 and P5 were isolated from the same fermenta- 
tion, but were not obtained in high enough yields to per- 
mit much structure work. 

H. S. Burton and E. P. Abraham, Biochem. J. 50 168 
(1951). (Isolation) 

H. S. Burton, E. P. Abraham and H. M. E. Cardwell, 
Biochem. J. 62 171 (1956). 

CIBA Lectures in Microbial Chemistry, E. P. Abraham, 
"Biochemistry of Some Peptide and Steroid Antibiotics," The 
cephalosporins, John Wiley and Sons, New York, 1957. pp. 
30-63. (A review) 



10 



Tropolone Acids 



The detailed biosynthetic origin of the tropolone acids remains 
obscure. Various suggestions have been made. One of these^' ^ 
proposed enlargement of the aromatic ring of 3,5-dihydroxy- 
phthalic acid, a known mold metabolite: 



HO 



COOH 




CH2O 



OH 



COOH 



HO 



HOCH; 



COOH 




lO] 



COOH 



OH 
HO COOH 



HO 




/"^X COOH 
O OH 




/ 



COOH 



OH 



Another^ proposed enlargement of an alicyclic ring in a Cg — C3 
type of intermediate from the shikimic acid route: 



CO— COOH 



HOOC CH2COCOOH HOOC CH 



[O] K^ [O] 
lO] 




10]^^[0] 
[O] 



IT. R. Seshadri, /. Sci. Ind. Research (India) 14B 248 (1955). 
^ R. Robinson, "The Structural Relations of Natural Products, 
Oxford Univ. Press, London, 1955. 

3 A. J. Birch, Fortschr. Chem. org. Naturstaffe 14 186 (1957^ 



Pfizer Handbook of Microbial Metabolites 



182 



COOH 




HO / 
O 



CO~COOH 



OH 
OH 




Labeling studies*- °' '^ show that acetate and formate are the 
primary precursors rather than glucose. Tanenbaum, Bassett 
and Kaplan found that stipitatic acid isolated from a Penicillium 
stipitatum culture grown on 1-C'^-glucose had an activity about 
five times as great as phenylalanine or tyrosine (shikimic acid 
route) isolated from the same culture. Richards and Ferretti 
grew Penicilliuvi aurantio-virens on media containing (a) 1-C^*- 
acetate, (b) 2-C'*-acetate and (c) 1-C'^-glucose. Puberulic acid 
and puberulonic acid were isolated, separated and degraded. 
Their results, in agreement with Bentley's where the same pre- 
cursors were used, indicate the incorporation of formate and 
acetate as follows: 



OH OH 



@— ^ + A - 
CH3— COOH formate 



HO 



1=0 HO-, 




/- 



® COOH 

Puberulic 
Acid 

That is, Cj, C-., C-, and C,s of the tropolones (as numbered in the 
puberulonic acid structure shown) are derived from the methyl 
carbon atom of acetate, while C^,, C4 and C,j are from the acetate 
carboxyl group carbon atom. The C- carbon atom of the trop- 
olones was shown by Bentley" to be derived from formate. 
The origin of the C9 carbon atoms present in puberulonic and 

* John H. Richards and Louis D. Ferretti, Biochem. and Biophys. 
Res. Comms. 2 107 (1960). 

5 Ronald Bentley, Biochim. et Biophys. Acta 29 666 (1958). 

" S. W. Tanenbaum, E. W. Bassett and M. Kaplan, Arch. Bio- 
chem. and Biophys. 81 169 (1959). 



183 Tropolone Acids 

stipitatonic acids remains undetermined. It, too, may arise 
from formate. A study has been made" of the enzymatic de- 
carboxylation of stipitatonic and puberulonic acids. A biochem- 
ical relationship was concluded by way of this enzyme, and the 
suggestion made that the intermediate tropolone precursors 
must be at least C., compounds, and that direct closure of an 
acyclic to a seven-membered ring structure must occur. 

The results of Richards and Ferretti seem to leave it an open 
question as to whether the tropolone ring is formed by direct 
cyclization of a long-chain acyclic compound or by expansion 
of a six-membered ring, and the exact nature of the interme- 
diate precursors of this interesting series of mold metabolites 
remains a mystery. 

372 Stipitatic Acid, CsHgOs, pale yellow plates, m.p. 282° (dec). 




COOH 

Penicillium stipitatum Thom 

J. R. Bartels-Keith, A. W. Johnson and W. I. Taylor, J. 
Chem. Soc, 2352 (1951). (Synthesis) 

Peter L. Pauson, Chem. Revs. 55 9 (1955). (A review of 
tropolones ) 

373 Puberulic Acid, CsH^O,;, nearly colorless plates, m.p. 318°. 

O OH 



COOH 



Penicillium puberulum Bainier, P. aurantio-virens 
Biourge, P. cyclopium-viridicatum and P. johannioli Za- 
leski 

R. E. Corbett, C. H. Hassall, A. W. Johnson and A. R. Todd, 
/. Chem. Soc, 1 (1950). 

Ronald Bentley and Clara P. Thiessen, Nature 184 552 (1959). 




Pfizer Handbook of Microbial Metabolites 1 84 

374 Stipitatonic Acid, C9H4O6, yellow crystals, m.p. 237° (dec). 

OH 



X 




\ 



HO 



Penicillium stipitatum Thorn 

W. Segal, Chem. and Ind., 1040 (1957). (Isolation) 

Kozo Doi and Yoshio Kltahara, Bull. Chem. Soc. Japan 31 
788 (1958). (Structure) 

W. Segal, Chem. and Ind., 1726 (1958). (Corrected struc- 
ture) 

375 Puberulonic Acid, C9H4O7, fine yellow needles, m.p. 298° (dec). 

O 



HO— (( 


r\. 




■^c 

II 





Penicillium johannioli Zaleski, P. cyclopimn-viridica- 
tum, P. puberulum Bainier and P. aurantio-virens Biourge 
See preceding reference. 

Gunhild Aulin-Erdtman, Chem. and Ind., 29 (1951). 
Idem., Acta Chem. Scand. 5 301 (1951). (Structure) 

376 Compound T, C10H8O4 or Ci,jHi„04, colorless crystals, m.p. 150°. 

This compound shows the typical tropolone spectrum, 
and it is apparently a new tropolone acid. 

Penicillium stipitatum 

S. W. Tanenbaum, E. W. Bassett and M. Kaplan, Arch. 
Biochem. and Biophys. 81 169 (1959). 



n 



Phenolic Substances 



a. PHENOLS AND PHENOL ETHERS (GENERAL) 

Phenolic substances are commonly encountered as microor- 
ganism metabolites. Besides the compounds listed in this chap- 
ter phenolic moieties are present in other structures such as the 
xanthones, altemariol, blastmycin, hygromycin, fulvic acid, cit- 
romycetin, atrovenetin, the tetracyclines, mycobactin, anthra- 
quinones and naphthoquinones. Benzoquinones are undoubt- 
edly oxidation products of phenolic precursors. 

Practically all of the phenolic materials in this section are 
mold metabolites. Perhaps that is because more isolation work 
has been done with fungi than with bacteria. It is evident that 
similar compounds are produced by bacteria, since 6-methyl- 
salicylic acid, a typical penicillium metabolite, also occurs as a 
moiety of mycobactin from Mycobacterium phlei. Also, 2,3- 
dihydroxybenzoic acid occurs as a moiety of a metabolite from 
Bacillus subtilis, and 2,6-dihydroxybenzoic acid as a part of 
pyoluteorin from Pseudomonas aeruginosa. It is interesting 
that these bacterial phenolic acids are conjugates of nitrogen- 
containing substances. 

The phenolic acid production of certain cultures has been 
studied in depth. Penicillium brevi-compactum, for example, 
has been found to produce the following: 
3,5-Dihydroxyphthalic Acid CsHgOg 
l-Carboxy-2,5-dioxyphenyl Acetyl Carbinol CioHioOg 
2,4-Dioxy-6-pyruvylbenzoic Acid CioHgOg 



Pfizer Handbook of Microbial Metabolites i86 

Mycophenolic Acid CiyHsoOg 
Another investigation^ in fact found a total of 11 different 
phenolic substances in a culture of this organism. In addition 
to the above were found a compound Ci„Hi„07, two derivatives 
of mycophenolic acid, two "intermediates between CigHio07 and 
CsH(;0(." and two reduction products of CioHioOj. 
The mold Penicillium griseofulvum produces: 

6-Methylsalicylic Acid CsHsOo 

Orsellinic Acid CsHs04 

Griseofulvin CiyHi^OfjCl 

Dechlorogriseofulvin Ci^HisOr, 

Bromogriseofulvin CiTH^^OgBr 

Gentisic Acid €711^,04 

Fulvic Acid C14H12OS 

Mycelianamide CooHosOriNo 
Another study- found three more unidentified phenolic sub- 
stances in this culture. 

A Penicillium patulum culture has been found^ to produce: 

Patulin C-H6O4 

Gentisaldehyde C^HgOg 

Gentisic Acid C7Hp,04 

Gentisyl Alcohol C7HhO;^ 

6-Methylsalicylic Acid CsHs04 

6-Formylsalicylic Acid CsHfi04 

3-Hydroxyphthalic Acid CsHgOg 

Pyrogallol C.jHjjO^ 

p-Hydroxybenzoic Acid C7H6O3 

Anthranilic Acid C7H7O2N 
Also an "aliphatic precursor of patulin" and a depside-like 
compound were detected but not entirely characterized. 

Many such families of metabolites can be assembled by ref- 
erence to the microorganism index. Studies such as those 
above facilitate the development of biosynthetic routes. For 
example, Bassett and Tanenbaum suggest the following inter- 
relationships among the Penicillium patulum phenolic metab- 
olites : 

1 Paul Godin, Antonie van Leeuxvenhoek J. Microbiol. Serol. 21 215 
(1955). 

- Paul Simonart and Renaat de Lathouwer, Zentr. Bakteriol., 
Parasitenk., Abt. II 110 339 (1957). 

2 E. Bassett and S. Tanenbaum, Experientia 14 38 (1958). 



i87 



Phenols and Phenol Ethers (General) 



Glucose 



COOH 




Shikimic Acid 



COOH 




OH 



Acetate 



CH3 



COOH 




HO !,„ OH 



COOH 



OH 



6-MethylsalicyIic 
Acid 



CHO 



COOH 





OH ^°°" in ^°°" 



Gallic Acid J p-Hydroxy- 6-Formylsalicylic 3-Oxyphthalic 
benzoic Acid Acid 

Acid 

i 

°" CHO 



COOH 



i 




HO 1^^ OH 

Pyrogallol 
OH 




OH 




OH OH 

CH2OH I CHO I COOH 





OH OH OH 

Gentisyl Gentisaldehyde Gentisic Acid 



Alcohol 



i 
OH 



OHC 




CHO 



/I 



OH 



Pfizer Handbook of Microbial Metabolites i88 

i 
OH COOH OH COOH 




Thus, the acetate and shikimic acid routes to aromatic com- 
pounds seem to be operating in a single microorganism. 

It was a kind of statistical consideration of the structures of 
natural products which led to the revival of the acetate hypothe- 
sis of biogenesis as applied to substances other than fatty acids. 
Phenolic compounds were particularly instrumental since 
the frequent occurrence of weta-hydroxyl groups (resorcinol 
and phloroglucinol types) was easy to recognize and challeng- 
ing to explain. The case first was stated clearly by Collie many 
years ago.* Lately Birch and others have developed a firm ex- 
perimental basis for the theory by isotopic labeling and chemi- 
cal degradation studies. 

Some phenolic compounds which have been shown in this 
way to be acetate-derived are : 

6-Methylsalicylic Acid^ 
Griseofulvin" 
Mycophenolic Acid" 
Emodin^ 

*John Norman Collie, Proc. Chem. Soc. 23 230 (1907); idem., }. 
Chem. Soc. 91 1806 (1907). 

•' A. J. Birch, R. A. Massy-Westropp and C. J. Moye, Australian J. 
Chem. 8 539 (1955). 

^ A. J. Birch, R. A. Massy-Westropp, R. W. Rickards and Herchel 
Smith, J. Chem. Soc, 360 (1958). 

" A. J. Birch, R. J. English, R. A. Massy-Westropp, M. Slaytor and 
Herchel Smith, ibid., 365 (1958); A. J. Birch, R. J. English, R. A. 
Massy-Westropp and Herchel Smith, ibid., 369 (1958). 

^ Sten Gatenbeck, Acta Chem. Scand. 12 1211 (1958). 



1 89 



Phenols and Phenol Ethers (General) 



Auroglaucin'' 
Helminthosporin^° 



COOH 
CH3 I OH 





6-Methylsalicylic 
Acid 



OHC 



CH3(CH=CH) 



CH3 



Griseofulvin 



CHa 




CH2— CH=C 



\ 



CHs 



OH 
Auroglaucin 



HOOC— CH2— CH2— C=CH— CH 



CH3O 




Mycophenolic Acid 



HO O OH 

Helminthosporin 




HO O OH 

Emodin 

Interesting details have been discovered. For example/ the 
methyl group attached to the aromatic ring in mycophenolic 
acid is furnished by methionine, probably at a relatively early 
stage in the synthetic sequence. The methoxyl methyl group 
also is furnished by methionine. The aromatic nucleus is ace- 
tate-derived, while mevalonic acid was shown to be a specific 

» A. J. Birch, J. Schofield and Herchel Smith, Chem. and Ind., 1321 
(1958). 

^° A. J. Birch, A. J. Ryan and Herchel Smith, /. Chem. Soc, 4773 
(1958). 



Pfizer Handbook of Microbial Metabolites i go 

and irreversible intermediate for the terpenoid side-chain. 
There was no incorporation of mevalonic acid into the aromatic 
nucleus. Mevalonic acid also was incorporated exclusively into 
the isopentene side-chain of auroglaucin. 

Both bacteria and fungi are able to hydroxylate aromatic 
rings, and the acetate pattern of alternate oxidation often is 
confused by further oxidations of this sort. 

Other details remain to be determined. The predominance 
of metabolites indicating derivation from an even number of 
acetate units has led to speculation concerning a four-carbon 
intermediate such as acetoacetate. Even larger intermediates 
have been proposed, such as orsellinic acids as precursors of 
anthraquinones." So far this possibility has not been ruled out 
in each case- by rigorous experimental evidence although there 
is an intuitive tendency to favor the simplest and most flexible 
unit and to apply the accumulated body of knowledge about in- 
termediary metabolism. The co-occurrence in a natural source 
of the anthraquinone and related phenanthrenequinone men- 
tioned in the introduction to the section on quinones is pre- 
sumptive evidence against orsellinic acid intermediates, since 
the two quinone molecules appear to be formed merely by a 
different mode of folding or arrangement on an enzyme surface 
of the same intermediate polyketomethylene chain. On the 
other hand the isolation of such orsellinic acids from isolated 
fungus members of lichens incapable of completing the anthra- 
quinone synthesis is interesting. 

The structural relationships (some obvious, others more ob- 
scure) among the mold products fulvic acid, citromycetin, fu- 
sarubin, purpurogenone, etc.^- ^' argue in favor of a flexible 
intermediate in the sense of a single polyketomethylene chain 
that could be folded and modified in various ways to give re- 
lated metabolites. Comparison of the structures of the lactone 
moieties of the macrolide antibiotics with those of the tetracy- 
clines (both classes of compounds produced by streptomycetes) 
also seems to point to intermediates of this type. While this is 
a good working hypothesis, such intermediates have not been 
isolated and in fact could not long exist in the free state. Per- 
haps eventually a better knowledge of enzymes will let us know 

" K. Aghoramurthy and T. R. Seshadri, J. Sci. Ind. Research 
(India) 13A 114 (1954). 

^^ F. M. Dean, R. A. Eade, R. A. Moubasher and A. Robertson, 
Nature 179 366 (1957). 

"W. B. Whalley, Chem. and Ind., 131 (1958). 



igi Phenols and Phenol Ethers (General) 

in more detail how such acetate-derived mold metabolites are 
formed, and why the chain lengths seldom exceed 14 to 18 
carbon atoms. 

The recent discovery and characterization of asterric acid, 
a mold metabolite in which two phenolic units are joined by an 
ether hnkage, have inspired the suggestion that the final phases 
of its biogenetic scheme may involve a geodin-like intermediate 
and sulochrin as follows: 

9^^^3 OH OCH3 

O I 



-^-^' 



HO— <v /^C— ^ \^CH 
COOCH3 OH 
Sulochrin 




OH 

COOCH3 COOH 
Asterric Acid 

The authors believe that the known occurrence of sulochrin 
and geodin as mold metabolites supports this argument." 

The transformation of sulochrin to dechlorogeodin, inciden- 
tally, is an example of intramolecular phenol coupling, a phe- 
nomenon discussed at greater length under Part b of this sec- 
tion. 

377 Pyrogallol, C,jH,;03, colorless crystals which turn brown in air, 
m.p. 133°. 




Penicillium patnlum 

E. W. Bassett and S. W. Tanenbaum, Biochim. et Biophys. 
Acta 28 247 (1958). 

"R. F. Curtis, C. H. Hassall and D. W. Jones, Chem. and Ind., 
1283 (1959). 



Pfizer Handbook of Microbial Metabolites 192 

378 p-Methoxytetrachlorophenol (Drosophilin A), C7H4O2CI4, yellow 
crystals, m.p. 118°, 




Drosophila subatrata (Batsch ex Fr. ) Quel. 

The yield was 100 mg. from 31 liters of culture solu- 
tion. 

Frederick Kavanagh, Annette Hervey and William J. Rob- 
bins, Proc. Natl. Acad. Sci. U. S. 38 555 (1952). 

379 p-Hydroxybenzoic Acid, CjHeOg, colorless crystals, m.p. 213°. 

COOH 



Penicillium patulum 

E. W. Bassett and S. W. Tanenbaum, Biochim. et Biophys. 
Acta 28 247 (1958). 

380 Protocatechuic Acid, C7H6O4, white or tan crystalline powder 
which darkens in air, m.p. —-200° (dec). Monohydrate 
from water. 



COOH 




Phycomyces blakesleeanus (sugar substrate) 
H. B. Schroter, Angew. Chem. 68 158 (1956). 

381 Gentisic Acid, C7H6O4, colorless crystals, m.p. 199°. 

°" COOH 




193 Phenols and Phenol Ethers (General) 

Penicillhim griseofulvum Dierckx, P. jenseni, P. diver- 
gens 

Harold Raistrick and Paul Simonart, Biochem. J. 27 628 
(1933). 

J. Barta and R. Mecir, Experientia 4 111 (1948). 

A. Brack, Helv. Chim. Acta 30 1 (1947). (Isolation) 

382 Gallic Acid, CjHgO^, colorless or pale tan crystals (Monohydrate 
from water), m.p. 225-250° (dec). 



COOH 




Phycomyces blakesleeanus (sugar substrate) 
Protocatechuic acid and another unidentified phenol 
also were shown to be present by paper chromatography. 
H. B. Schroter, Angew. Chem. 68 158 (1956). 

383 Gentisyl Alcohol, CyHgOg, colorless crystals, m.p. 100°. 

OH 

CH2OH 




Penicillium patulum Bainier, P. divergens Bainier and 
Sartory 

A. Brack, Helv. Chim. Acta 30 1 (1947). (Isolation) 

B. G. Engel and W. Brzeski, ibid. 30 1472 (1947). 
J. Barta and R. Mecir, Experientia 4 277 (1948). 

384 2,5-Dihydroxyphenylglyoxylic Acid, CgHeOj, yellow needles, m.p. 
141°. 



COCOOH 




Polyporus tumulosus Cooke (artificial medium) 
Oxalic acid, homoprotocatechuic acid and 2,4,5-trihy- 
droxyphenylglyoxylic acid are produced in the same cul- 
ture. 



Pfizer Handbook of Microbial Metabolites 194 

Otto Neubauer and L. Flatow, Hoppe-Seyler's Zeitschrift 
fiirphysiol. Chem. 52 375 (1907). 

G. F. J. Moir and B. F. Ralph, Chem. and Ind., 1143 (1954). 

385 3-Hydroxyphthalic Acid, CsHeO^, colorless crystals m.p. : anhy- 
dride formation near 150°, melting 166°. Sublimes. 



COOH 



COOH 



Penicillium islandicum, P. patulum 
A yield of only 1-2 mg. per liter was obtained. 
Sten Gatenbeck, Acta Chem. Scand. 11 555 (1957). 
E. W. Bassett and S. W. Tanenbaum, Experientia 14 38 
(1958). 

386 3,5-Dihydroxyphthalic Acid, CgHgOg, colorless prisms, m.p, 188° 
(resolidifying at 206°). 

HO COOH 





OH <=°°" 

Penicillium brevi-compactum Dierckx 

Albert E. Oxford and Harold Raistrick, Biochem. J. 26 1902 

(1932). (Isolation) 

John Howard Birkinshaw and Arthur Bracken, J. Chem,. 

Soc, 368 (1942). (Synthesis) 

387 2,4,5-Trihydroxyphenylglyoxylic Acid, CgHgOe, bright red prisms, 
m.p. 193°. 

O 

OH II 

C— COOH 




Polyporus tumulosus (artificial medium) 
Homoprotocatechuic acid and oxalic acid were present 
in the same culture. 



195 Phenols and Phenol Ethers (General) 

B. J. Ralph and Alexander Robertson, /. Chem. Soc, 3380 
(1950). 

388 2,6-Dihydroxyacetophenone, CsHhOj, yellow needles, m.p. 154- 
158°. 

OH 
I COCH3 



OH 

Daldinia concentrica 

D. C. Allport and J. D. Bu'Lock, J. Chem. Soc, 654 (1960). 

389 6-Methylsalicylic Acid (2,6-Cresotic Acid, 3-Hydroxy-5-toluic 
Acid, 6-Hydroxy-2-methylbenzoic Acid), CgHgOy, colorless 
needles, m.p. 170°. 

COOH 
CH3, I ,OH 



Penicillium griseofulvum Dierckx, P. flexuosiim, P. pat- 
ulum Bainier, P. urticae 

Winston Kennay Anslow and Harold Raistrick, Biochem. J. 
25 39 (1931). 

E. W. Bassett and S. W. Tanenbaum, Experientia 14 38 
(1958). 

390 p-Hydroxyphenylacetic Acid, CgHgO^, colorless crystals, m.p. 
148° (subl.). 

HO—/ \— CH2COOH 



Hypochnus sasakii Shirai (Corticium sasakii, Pellicu- 
laria sasakii) 

Ysu Shik Chen, Bull. Agr. Chem. Soc. (Japan) 22 136 
(1958). 

391 Homoprotocatechuic Acid, C8H8O4, colorless plates, m.p. 128.5°. 

CHoCOOH 




Pfizer Handbook of Microbial Metabolites 196 

Polyporus tumulosus (artificial medium) 
B. J. Ralph and Alexander Robertson, J. Chem. Soc, 3380 
(1950). 

392 Orsellinic Acid, C8H8O4, colorless crystals, m.p. ( Monohydrate ) 
176°. 

?"^ COOH 



HO OH 

Penicillium griseofulvum, Chaetomium cochlioides 

L. Reio, /. Chromatography 1 338 (1958). 

Klaus Mosbach, Zeitschr. Naturforsch. 14b 69 (1959). 

393 Compound D, CoHgO,^, cream-colored prisms, m.p. 259° (dec). 

A meta diphenol with a carboxyl group para to a hy- 
droxyl and an aldehyde group ortho to a hydroxyl. 

Paecilomyces victoriae V. Szilvinyi 

Ustic acid, dehydroustic acid and 4,6-dihydroxy-3-meth- 
oxyphthalic acid were isolated from the same culture. 

V. C. Vora, J. Sci. Ind. Research (India) 13B 842 (1954). 

394 Flavipin, CgHgOg, pale yellow light-sensitive rods, m.p. 233° 

(dec). 




CHO 
CHO 

Aspergillus flavipes (Bainier and Sartory) Thom and 
Church, A. terreus Thom 

P. Rudman, "Metabolic Products of A. fiavipes, A. terreus 
and Certain Other Molds," Doctoral Thesis, Univ. of London, 
London, 1955. 

H. Raistrick and P. Rudman, Biochem. J. 63 395 (1956). 

395 4,6-Dihydroxy-3-methoxyphthalic Acid, CgHgO^, colorless prisms, 
m.p. 193°. 

HO COOH 




OCH.^°°" 



1 97 Phenols and Phenol Ethers (General) 

Paecilomyces victoriae V. Szilvinyi 

Ustic acid, dehydroustic acid and another incompletely 
characterized phenolic acid were isolated from the same 
culture. 

V. C. Vora, /. Sci. Ind. Research (India) 13B 842 (1954). 

396 2,3-DihydroxybenzoyIglycine, C9H9O5N, colorless needles, m.p. 
210°. 



HO OH_ 

\ / O 



-C— NH— CH2— COOH 




Bacillus subtilis (iron-deficient medium) 
Coproporphyrin and succinic acid were also produced. 
Takeru Ito and J. B. Neilands, /. Am. Chem. Soc. 80 4645 
(1958). 

397 8-Hydroxy-3-niethylisocoumarin, CjoHgOg, colorless needles, m.p. 
99°. 

O 

II 
HO ,C 



O 

Marasmius ramealis 

Gerd Benz, Arkiv for Kemi 14 511 (1959). 

398 2,4-Dioxy-6-pyruvylbenzoic Acid, CioHgOg, fine colorless crystals, 
m.p. 125-135°. 

HO 

\ 00 

^ ^C— C— CH3 

HO COOH 

Penicillium brevi-compactum (syn. P. stoloniferum 
Thorn) 

Percival W. Clutterbuck, Albert E. Oxford, Harold Ralstrick 
and Geo. Smith, Biochem. J. 26 1441 (1932). (Isolation) 

Albert E. Oxford and Harold Raistrick, ibid. 27 634 (1933). 




Pfizer Handbook of Microbial Metabolites 198 

399 Mellein (Ochracin), C10H10O3, colorless prisms, m.p. 58°, [a]D 

-124.86° ([aW- -108.i5° in chloroform). 

O 

II 

^CH— CH3 
CH2^ 

Aspergillus melleus Yugawa, A. ochraceus 

Eijiro Nishikawa, /. Agr. Chem. Soc. Japan 9 772 (1933). 
(Isolation) (Chem. Abstr. 28 2751) 

Teijiro Yabuta and Yusuke Sumiki, ibid. 9 1264 (1933). 
(Isolation) (Chem. Abstr. 28 2350) 

John Blair and G. T. Newbold, J. Chem. Soc, 2871 (1955). 
(Structure) 

It is interesting to note that a similar compound : 

O 

OH I 

/ > 

^CH— CH3 
CH3O \h/ 

has been isolated from carrots which had developed a 
bitter taste during cold storage. 

Ernest Sondheimer, J. Am. Chem. Soc. 79 5036 (1957). 
(Isolation") 

400 3,5-Dimethyl-6-oxyphthalide, Ci^HioOg, colorless needles, m.p. 

156-158°. 




Penicillium gladioli 



1 99 Phenols and Phenol Ethers (General) 

H. Raistrick and D. J. Ross, Biochem. J. 50 635 (1952). 
401 Quadrilineatin, CioHn,04, colorless needles, m.p. 172° (dec.)- 

HO CHO 




Aspergillus quadrilineatus Thorn and Raper 
J. H. Birkinshaw, P. Chaplen and R. Lahoz-Oliver, Biochem. 
J. 67 155 (1957). 

402 l-Carboxy-2,5-dioxy benzyl Methyl Ketone, Ci„HioO.-,, large dia- 

mond-shaped crystals, m.p. 152-156° (dec), remelting 
at 220-230°. 

\ O 

^ \— CH2— C— CH3 

HO COOH 

Penicillium brevi-compactum (syn. P. stoloniferum 
Thom) 

Percival W. Clutterbuck, Albert E. Oxford, Harold Raistrick 
and Geo. Smith, Biochem. }. 26 1441 (1932). 

Albert E. Oxford and Harold Raistrick, ibid. 27 634 (1933). 

403 l-Carboxy-2,5-dioxyphenyl Acetyl Carbinol, CioHjoOe, colorless 

rhombs, m.p. 202-204° (dec). 

"\ OH O 

^ V-CH— C— CH3 

HO COOH 

Penicillium brevi-compactum (syn. P. stoloniferum 
Thom) 

Percival W. Clutterbuck, Albert E. Oxford, Harold Raistrick 
and Geo. Smith, Biochem. J. 26 1441 (1932). 



Pfizer Handbook of Microbial Metabolites 200 

404 2,6-Dihydroxybutyrophenone, C10H12O3, yellow needles, m.p. 
116.5-118°. 

OH 
1 COCH2CH2CH3 




OH 

Daldinia concentrica 

D. C. Allport and J. D. Bu'Lock, /. Chem. Soc, 654 (1960). 

405 Clavatol, C10H10O3, colorless plates, m.p. 183°. 

°" COCH. 




Aspergillus clavatus 

Occurs as a minor product with patulin in this culture. 

F. Bergel, A. C. Morrison, A. R. Moss and H. Rinderknecht, 
J. Chem. Soc, 415 (1944). (Isolation) 

C. H. Hassan and A. R. Todd, ibid., 611 (1947). (Struc- 
ture) 

406 Sparassol, CiuHjoO^, colorless microcrystals, m.p. 67°. 

CH3 

I COOCH3 



HO OCH3 

Sparassis ramosa, Evernia pnaiastri 

John Stenhouse, Ann. 68 55 (1848). 

Emil Fischer and Kurt Hoesch, ibid. 391 347 (1912). 
(Structure) 

Richard Falck, Ber. 56B 2555 (1923). 

E. Wedekind and K. Fleischer, ibid. 56B 2556 (1923). 
(Structure) 

Ernst Spath and Karl Jeschki, ibid. 57A 471 (1924). 

407 N-Acetyltyramine, CioHi^OoN, colorless crystals, m.p. 135° (s. 
128°). 

HO—/ V-CH,CH,NHCOCH3 



Streptomyces griseiis (Krainski) Waksman et Henrici, 
Mycobacteriiiin tuberculosis 



201 Phenols and Phenol Ethers (General) 

J. Comin and W. Keller-Schierlein, Helv. Chim. Acta 42 
1730 (1959). 

Yutaka Shirai, Kekkaku (Tuberculosis) 30 628 (1955). 
(Chem. Abstr. 50 5839g) 

408 Gladiolic Acid, CnHjoOg, colorless needles, m.p. 158-160°. 





CHO ="° 



Penicillium gladioli McCuU. and Thorn 

Yield 300 mg. per liter. 

Besides dihydrogladiolic acid and 3,5-dimethyl-6-oxyph- 
thalide, a third "contaminant," C11H10O4 (a lactone), was 
present in the culture. 

John Frederick Grove, Biochem. J. 50 648 (1952). (Struc- 
ture) 

P. W. Brian, P. J. Curtis and H. G. Hemming, /. Gen. 
Microbiol. 2 341 (1948). (Isolation) 

409 Cyclopaldic Acid, CnHifjOg, colorless needles, m.p. 224° (subl.). 





CHO ^"° 



Penicillium cyclopium Westling 

J. H. Birkinshaw, H. Raistrick, D. J. Ross and C. E. Stlck- 
ings, Biochem. J. 50 610 (1952). 

410 Dihydrogladiolic Acid, C11H12O5, colorless crystals, m.p. 135° 
(dec). 

O 

°'="'COOH CH, ?="•! 

o 





CHO I CH 

CH2OH CH2OH I 

OH 



Penicillium gladioli 



Pfizer Handbook of Microbial Metabolites 202 

H. Raistrick and D. J. Ross, Biochem. J. 50 635 (1952). 
411 Cyclopolic Acid, CnHioOg, colorless plates, m.p. 147° (dec.)- 

O 
CH3 ?^"^ COOH CH 





HO I CHO HO 

CH2OH CH2OH I 



Penicillium cyclopium 

J. H. Birkinshaw, H. Raistrick, D. J. Ross and C. E. Stick- 
ings, Biochem. J. 50 610 (1952). 

412 Ustic Acid, C11H12O7, colorless crystals, m.p. 169° (dec). 

°" COOH 




OCH3 CH-C^H, 



OH O 



Aspergillus ustus, Paecilomyces victoriae, Ustilago zeae 

H. Raistrick and C. E. Stickings, Biochem. }. 48 53 (1951). 
Yield about 0.5 g. per liter. 

V. C. Vora, /. Sci. Ind. Research (India) 13B 842 (1954). 

Occurred together with dehydroustic acid, 4,5-dihy- 
droxy-3-methoxyphthalic acid and a fourth compound, 
C,,Hs05, m.p. 259°; an m-dihydroxyphenol with a carbonyl 
group and a carboxyl group. 

413 Radicinin,* CjoHioOr,, optically active crystals. 
Proposed Structure: 

O 
O^ / 



HO 



CH3 



I C— CH=CH2 



H 



O 



Stemphylium radicinum 

D. D. Clarke and F. F. Nord, Arch. Biochem. and Biophys. 
59 269-284 (1955). 
See also entry 871. 



203 Phenols and Phenol Ethers (General) 

414 Alternariol, Ci4Hj„0-,, colorless needles, m.p. 350° (dec.) 

and 

415 Alternariol Methyl Ether, CisHjoO.-,, colorless needles, m.p. 267° 

(dec). 



Q / OH 

CH3 OH ^ 

The methyl ether is at one of the positions indicated. 
Alternaria tenuis 

The yield was about V2 g. per Uter. 

H. Raistrick, C. E. Stickings and R. Thomas, Biochem. J. 
55 421 (1953). 

416 Altertenuol, C14H10O6, buff -colored rods, m.p. 284° (dec. and 

subl.). 

Forms a triacetate and a trimethyl derivative. Prob- 
ably related to alternariol. 

Alternaria tenuis 

T. Rosett, R. H. Sankhala, C. E. Stickings, M. E. U. Taylor 
and R. Thomas, Biochem. }. 67 390 (1957). 

417 Sorbicillin, Ci4H|,.03, orange plates, m.p. 113° (remelting at 

129°). 

CH. ?" ? 




"° CH, 

Penicillium notatum Westhng 

Donald J. Cram and Max Tishler, /. Am. Chem. Soc. 70 
4238 (1948). (Isolation from Clinical Sodium Penicillin) 

Donald J. Cram, ibid. 70 4240 (1948). (Structure) 

Besides sorbicillin several other compounds were iso- 
lated from careful investigation of a sample of early clini- 
cal sodium penicillin. In view of the source it is hard to 
say which of these may be considered true metabohtes. 
The other compounds were: 
Tiglic Acid, C.^HsOo, m.p. 63° 

d-a-Methylbutyric Acid, C5H10O2 b.p. 175°, [ajn'" +15.2° 
Furoic Acid, m.p. 129° 



Pfizer Handbook of Microbial Metabolites 204 

/5-Indole acetic Acid, m.p. 167° 
Phenylacetic Acid, m.p. 76° 
2-Decenedioic Acid, C10H16O4, m.p. 172° 
Pigment I (/?-Penetrin), m.p. 207°. 

^-Penetrin is identical with an alkaline hydrolysis prod- 
uct of penetrinic acid, a metabolite of P. notatum reported 
earlier 

Pigment II, CioHuO,;N, orange prisms, m.p. 105°, N.E. 
indicates a dicarboxylic acid. Optically inactive. Nega- 
tive FeClg test. Decolorizes permanganate. Decolorized 
by sodium hydrosulfite and apparently reduced to a hydro- 
quinone, m.p. 129°. 

Frank H. Stodola, Jacques L. Wachtel, Andrew J. Moyer 
and Robert D. Coghill, J. Biol. Chem. 159 67 (1945). 

418 Dehydroaltenusin, CigHiaOe, yellow needles, m.p. 189° (dec). 

An acidic compound probably related to altenusin. 
Alternaria tenuis 

T. Rosett, R. H. Sankhala, C. E. Stickings, M. E. U. Taylor 
and R. Thomas, Biochem. }. 67 390 (1957). 

419 Altenusin, Ci-,Hi406, colorless prisms, m.p. 202° (dec). 

An acidic compound which forms a tetramethyl deriva- 
tive. Probably related to alternariol. 

Alternaria tenuis 

T. Rosett, R. H. Sankhala, C. E. Stickings, M. E. U. Taylor 
and R. Thomas, Biochem. J. 67 390 (1957). 

420 Altenuic Acid I, C15H14O8, colorless needles, m.p. 183°, second 

m.p. 224-230° (dec). 

A dibasic acid probably related to alternariol. 

Alternaria tenuis 

T. Rosett, R. H. Sankhala, C. E. Stickings, M. E. U. Taylor 
and R. Thomas, Biochem. J. 67 390 (1957). 

421 Altenuic Acid II, Ci-,Hi408, colorless plates, m.p. 245° (dec). 

A dibasic acid probably related to alternariol. 
Alternaria tenuis 

T. Rosett, R. H. Sankhala, C. E. Stickings, M. E. U. Taylor 
and R. Thomas, Biochem. J. 67 390 (1957). 

422 Altenuic Acid III, C].,Hi40s, colorless prisms, m.p. 198-202°, 

second m.p. 225° (dec). 

A dibasic acid probably related to alternariol. 

Alternaria tenuis 

T. Rosett, R. H. Sankhala, C. E. Stickings, M. E. U. Taylor 
and R. Thomas, Biochem. }. 67 390 (1957). 



205 Phenols and Phenol Ethers (General) 

423 Penitrinic Acid, C15H17O5N, pale yellow bars, m.p. 217-223° 

(dec), [ali)-'^ —549° (in dimethylformamide). 

Similar in structure to sorbicillin. The two pigments 
occur together. 

Penicillhnn notatum Westling 

Frank H. Stodola, Jacques L. Wachtel, Andrew J. Moyer and 
Robert D. Coghill, J. Biol. Chem. 159 67 (1945). 

Kei Arima, Kazuo Kamagata and Hideo Nakamura, /. Agr. 
Chem. Soc. Japan 27 389 (1953). (Structure work) 

424 d,Z-Erdin, CigHioOyCL, yellow crystals, m.p. 210-212°. 

CH3 f OCH3 

' OH ^COOH 

Aspergillus terreiis Thom 

Erdin occurs naturally as the racemate although the 
closely related geodin, which is present in the same cul- 
ture, is the d-isomer. 

Harold Raistrick and George Smith, Biochem. J. 30 1315 
(1936). (Isolation) 

D. H. R. Barton and A. I. Scott, /. Chem. Soc, 1767 (1958). 
( Structure ) 

425 Curvularin, CieHo.jOg, colorless crystals, m.p. 206°, [ah^^ —36.3° 

(c 3.8 in ethanol). 



OH 



O 

il 

C(CH2)5 — CH— CH3 




/ 
HO CH2 



Curvularia sp. 

The yield was 0.40 to 0.48 g. per liter of culture broth. 
A second compound CieHigO.r;, m.p. 224.5°, [ajn'^ -83°, 
(also phenolic) was isolated from the same culture. 

C. Calam (Imperial Chemical Industries), unpublished. 
(Isolation) 

O. C. Musgrave, /. Chem. Soc, 4301 (1956). (Isolation) 



Pfizer Handbook of Microbial Metabolites 206 

Idem., ibid., 1104 (1957). 

A. J. Birch, O. C. Musgrave, R. W. Rickards and Herchel 
Smith, ibid., 3146 (1959). (Structure) 

426 d-Geodin, C17H12O7CI2, yellow crystals, m.p. 228-231°, [ah 
+ 140° (c 0.80 in chloroform). 




OH ^COOCHs 

Aspergillus terreus Thorn 

Harold Raistrick and George Smith, Biochem. J. 30 1315 
(1936). (Isolation) 

D. H. R. Barton and A. I. Scott, /. Chem. Soc, 1767 (1958). 
( Structure ) 

427 Geodoxin, Ci^HioOsCL, yellow needles, m.p. 216° (dec). 

CH3 

.0; 



Aspergillus terreus Thom 

C. H. Hassall and T. C. McMorris, J. Chem. Soc, 2831 
(1959). 

428 Sulochrin, Ci^HjeO^, colorless crystals, m.p. 262°. 

OCH3 
JHO\ 






Oospora sulfurea-ochracea 

Hidejiro Nichikawa, Bull. Agr. Chem. Soc. (Japan) 12 47 
(1936). 

Idem., J. Agr. Chem. Soc. Japan 13 1 (1937). 



207 Phenols and Phenol Ethers (General) 

Idem., Bull. Agr. Chem. Soc. (Japan) 16 97 (1940). 

429 Geodin-like Antibiotic, yellow crystals, ni.p. 229° (subl. 175° at 

3 mm.), [all.'-" +175° (in chloroform). 

The chlorine-containing part of the molecule is the same 
as that of geodin as shown by hydrolysis fragments. Other 
chemical and physical properties are similar to those of 
geodin. 

Aspergillus fiavipes 

Paul Delmotte, Julia Delmotte-Plaquee and Rene Bastin, /. 
Pharm. Belg. 11 200 (1956). 

430 Griseofulvin (Fulvicin, Grisovin) CiYHi^OfiCl, colorless crystals, 

m.p. 220°, [a]„-^ +337° (c 1.0 in acetone). 



OCH3O 




OCH3 



^>c c=o 

CH3O i. CH— CH2 

CH3 

Penicillium griseofulvum Dierckx, P. patidum, P. cd- 
bidum Sopp., P. raciborskii Zal., P. vielinii Thom, P. ur- 
ticae Bain., P. raistrickii, P. janczeivski Zal. (P. nigricans 
Thom and Bainier), Carpenteles brefeldianum Dodge 
(Shear) 

Albert Edward Oxford, Harold Raistrick and Paul Slmonart, 
Biochem. J. 33 240 (1939). (Isolation) 

J. C. McGowan, Trans. Brit. Mijcol. Soc. 29 188 (1946). 

P. J. Curtis and J. F. Grove, Nature 160 574 (1947). 

P. W. Brian, P. J. Curtis and H. G. Heming, Brit. Mycol. 
Soc. Trans. 32 30 (1949). 

John Frederick Grove, Doreen Ismay, J. MacMillan, T. P. C. 
MulhoUand, M. A. Thorold Rogers, Chem. and Ind., 219 
(1951). (Structure) 

Idem., J. Chem. Soc, 3958 (1952). 

John Frederick Grove, J. MacMillan, T. P. C. MulhoUand 
and M. A. Thorold Rogers, ibid., 3949, 3977 (1952). (Struc- 
ture) 

John Frederick Grove, J. MacMillan, T. P. C. MulhoUand 
and (Mrs.) J. Zealley, ibid., 3967 (1952). 

T. P. C. MulhoUand, ibid., 3987, 3994 (1952). 

A. J. Birch, R. A. Massy-Westropp, R. W. Rickards and 
Herchel Smith, Proc. Chem. Soc, 98 (1957). (Biosynthesis) 




Pfizer Handbook of Microbial Metabolites 208 

431 Bromogriseofulvin, Ci7Hi706Br, colorless crystals, m.p. 204°. 

O OCH3 

OCH3 II I 

I C C=CH 

\ / \ 

c c=o 

/ \ / 

CH3O I O CH — CH2 

Br I 

CHs 

On the proper medium bromogriseofulvin generally can 
be produced by the same molds which produce griseoful- 
vin. 

J. MacMillan, J. Chem. Soc, 2585 (1954). (Isolation) 

432 Dechlorogriseofulvin, Ci^HigOg, colorless needles, m.p. 179- 

181°, [alo" +390° (c 1 in acetone). 

OCH3 

?'"7 C=CH 

CH3O CH— CH2 

CH3 

Penicillium griseofulvum Dierckx, P. janczewski Zal. 

J. MacMillan, Chem. and Ind., 719 (1951). 

Idem., J. Chem. Soc, 1697 (1953). 

D. H. R. Barton and T. Bruun, /. Chem. Soc, 603 (1953). 

433 Mycophenolic Acid, C17H20O6, colorless needles, m.p. 141°. 

CH3 

I 
HOOCCH2CH2— C=CH— CH 



CH3O 




_rM>0 



Penicillium brevi-compactum Dierckx 

C. L. Alsberg and O. F. Black, Bull. U. S. Bur. PI. Ind., 
No. 270 (1913). (Isolation) 

Percival Walter Clutterbuck, Albert Edward Oxford, Harold 
Raistrick and George Smith, Biochem. }. 26 1441 (1932). 



209 Phenols and Phenol Ethers (General) 

J. H. Birkinshaw, A. Bracken, E. N. Morgan and H. Rai- 
strick, ibid. 43 216 (1948). 

J. H. Birkinshaw, H. Raistrick and D. J. Ross, Biochem. J. 
50 630 (1952). (Structure) 

434 Xanthocillin-X, CisHioOoNo, yellow crystals, m.p. --200° (dec). 

HO— /^>— CH=C C=CH— /^>— OH 

N N 



Penicillium notatum Westling 

Xanthocillin constitutes about 70% of a mixture con- 
taining a second constituent, xanthocillin-Y. 

W. Rothe, Deutsche Med. Wochenschr. 79 1080 (1954). 
(Isolation) 

I. Hagedorn and H. Tonjes, Pharmazie 11 409 (1956). 
(Structure) 

Use Hagedorn, Ulrich Eholzer and Arthur Luttringhaus, 
Chem. Ber. 93 1584 (1960). (Experimental work) 

435 Auroglaucin, C19H22O3, orange-red crystals, m.p. 153°, 

CH3 

OHC °" 




CH,(CH=CHI. .^ 
On 

Aspergillus glaucus, A. mangini, other aspergilli 

H. Raistrick, Robert Robinson and A. R. Todd, J. Chem. Soc, 

80 (1937). 

Adolfo Quilico, Cesare Cardani and Luigi Panizzi, Atti 

accad. nazl. Lincei Rend., Classe sci. fis.. Mat. e nat. sci. 14 358 

(1953). (Structure) 

436 Flavoglaucin, C19H2SO3, pale yellow crystals, m.p. 103°. 

CH3 

OHC °" 



CHslCHzle 




Pfizer Handbook of Microbial Metabolites 



2IO 



Aspergillus glaucus, other aspergilli 

H. Raistrick, Robert Robinson and A. R. Todd, /. Chem. 
Soc, 80 (1937). 

Adolfo Quilico, C. Cardani and G. Stagno d'Alcontres, Gazz. 
chim. ital. 83 754 (1953). (Structure) 

437 Picrolichenic Acid, Co-H.^hOy, colorless crystals, m.p. 178° (dec). 
Proposed structure: 



CH3O 




CH3CH2CH2CH2CH2 



OH 
COOH 

CH2CH2CH2CH2CH3 



Pertusaria amara (Ach.) Nyl., Variola amara (Ach.) 

The yield was 5-10 S^ of the dry weight of the lichen. 

H. Erdtman and C. A. Wachtmeister, Chem. and Ind., 1042 
(1957). 

Carl Axel Wachtmeister, Acta Chem. Scand. 12 147 (1958). 
(Structure) 

438 a-Tocopherol (Vitamin E), Co^H-.^Oo^ viscous oil, b.p. 200-220° 
(0.1 mm.), Ud'' 1.5045, U.V. max. 294 m/x. 



HO 



CHs 



CH3 



CH; 



/^ (CHo)3— CH— (CHo)3— CH— (CHsls— CH— CHs 

^O^ CH3 CH3 CH3 

CH3 



Identified in about a dozen varieties of chlorophyll-con- 
taining bacteria by paper chromatographic comparisons. 
(Not isolated.) 

J. Green, S. A. Price and L. Gare, Nature 184 1339 (1959). 

439 Chartreusin (Antibiotic X-465A), C;^2H320l4, greenish yellow 
crystals, m.p. 184-186°, [aW^ +132° ±6° (c 0.2 in pyri- 
dine). 



211 Phenols and Phenol Ethers (General) 

Proposed Structure: 



D-Digitalose — D-Fucose 




CH3 
D-Digitalose 

Streptomyces chartreusis and probably other Strepto- 
myces spp. 

Byron E. Leach, Kenneth M. Calhoun, LeRoy E. Johnson, 
Charlotte M. Teeters and William G. Jackson, J. Am. Chem. 
Soc. 75 4011 (1953). (Isolation) 

K. M. Calhoun and L. E. Johnson, Antibiotics and Chemo- 
therap7j 6 294 (1956). 

Julius Berger, L. H. Sternbach, R. G. Pollock, E. R. LaSala, 
S. Kaiser and M. W. Goldberg, /. Am. Chem. Soc. 80 1636 
(1958). 

L. H. Sternbach, S. Kaiser and M. W. Goldberg, ibid. 80 
1639 (1958). 

E. Simonitsch, W. Eisenhuth, O. A. Stamm and H. Schmid, 
Helv. Chim. Acta 43 58 (1960). (Structure) 

440 Chartreusin-like Antibiotic, C32H34O14, m.p. 186°. 
A weakly acidic glucoside. 
Streptomyces sp. 

F. Arcamone, F. Bizioli and T. Scotti, Antibiotics and 
Chemotherapy 6 283 (1956). 



Pfizer Handbook of Microbial Metabolites 212 

b. DEPSIDES AND DEPSIDONES 

Lichens are symbiotic partnerships of fungi and algae. While 
this slow-growing combination is visible without the aid of 
lenses, the extractable metabolites so resemble those of micro- 
organisms that they are included in this listing for comparison. 

Lichens and the fruiting bodies of the higher fungi were long 
used in folk medicine in the damp northern lands where they 
are prominent members of the flora. It was only natural, then, 
that the tool of organic chemistry was applied at an early date 
in these locations to elucidate the structures of their metabolites. 
Thus, historically, a large body of knowledge on such structures 
existed long before systematic work was begun on the fungi and 
streptomycetes, which have been so much more rewarding to 
modem medicine. 

Depsides, e.g. microphyllic acid and olivetoric acid, frequently 
contain aliphatic side-chains attached to their phenolic rings. 
The fact that these invariably consisted of an uneven number 
of carbon atoms was soon recognized and used as a rule in 
structure determinations. It was considered a curious phenom- 
enon until it became apparent that such molecules are particu- 
larly obvious examples of derivation from acetate. 

Certain lichen metabolites, for example some of the anthra- 
quinone pigments, have been found also in fungi. Moreover, 
some of the fungal partners have been isolated from lichens and 
grown alone in pure culture. In a few such cases the same 
metabolites have been isolated which are produced by the part- 
nership itself . Examples are the anthraquinones phy scion (pa- 
rietin) and rhodocladonic acid, the dibenzofurans usnic and 
didymic acids, as well as pulvic anhydride (stictaurin) and 
the nidulins.^ - * 

In contrast there is evidence that depsides and depsidones 
cannot be produced by the isolated fungus partner, but are the 
unique products of a collaborative effort.* In the work just 
cited it was found that the fungal components of various cla- 

^ E. Thomas, Beitr. 2. Kryptogamenfiora der Schweiz 9 1 (1939). 

-Hempstead Castle and Flora Kubsch, Arch. Biochem. 23 158 
(1949). 

^ F. M. Dean, A. D. T. Erni and Alexander Robertson, /. Chem. 
Soc, 3545 (1956). 

* Dieter Hess, Z. Naturforsch. 14b 345 (1959). 



213 



Depsides and Depsidones 



donia, parmelia and placodium species, grown alone in pure 
culture, produced no depsides nor depsidones. Orsellinic and 




COOH 



R=H=Orsellinic Acid 



R^CHO=Haematommic Acid 



haematommic acids, simpler moieties which could not be shown 
to be present as such in the parent lichens, were isolated. This 
could indicate that these phenols are precursors, and that the 
algae are necessary to effect coupling as well as final, charac- 
teristic modifications. It is interesting that orsellinic acid (q.v. ) 
has been isolated recently from other fungus cultures. Phenolic 
acids of this sort are obviously acetate-derived. 

Depsidones probably are formed by phenol coupling of the 
depsides. Phenol coupling (phenol dehydrogenation) is un- 
doubtedly a widespread phenomenon among natural products. 
It involves the removal of one electron from the phenol with 
formation of a phenol-free radical. Such radicals are relatively 
stable due to the resonance possibilities. In complex natural 
products such phenol radicals can form new bonds by intra- 
molecular attack. Thus the formation of a depsidone (in this 
case protocetraric acid) from a depside might be represented 
as follows: 



COOH 




COOH 




COOH 



Another example of intramolecular carbon-oxygen coupling was 
noted earlier in this chapter in the formation of the geodin, 
griseofulvin type of skeleton. 

Carbon-carbon bonds can be formed similarly (by coupling 



Pfizer Handbook of Microbial Metabolites 214 

of the ortho and para resonance isomers of the phenol-free 
radical). Biphenyl, binaphthyl and fois-anthraquinone skele- 
tons might be formed in this way. 

A combination of the two types of bond formation (i.e. first 
an intermolecular carbon-carbon coupling followed by an intra- 
molecular oxygen-carbon coupling) probably occurs in the bio- 
synthesis of compounds such as the dibenzofurans, etc. 

More thorough considerations of phenol coupling as a bio- 
synthetic process have been published. ' '' 

In vitro couplings of phenolic compounds have been accom- 
plished in the laboratory, by using simple electron acceptors 
such as molecular oxygen or ferric chloride, and natural prod- 
ucts have been prepared in this way. Yields under such condi- 
tions are generally low, and the orienting influence of the en- 
zyme surface seems to be required for real efficiency. 

Referencing of this section is lean because of the very thor- 
ough existing work.' In general the final structure determina- 
tion or synthesis is mentioned. 

441 Diploicin, C16H10O5CI4, colorless crystals, m.p. 232°. 

^' OCH. 




CI '-' CH3 

Buellia canescens (Dicks.) DeNot. 

Thomas J. Nolan, Joseph Algara, Eugene P. McCann, Wm. 
A. Manahan and Niall Nolan, Sci. Proc. Roy. Dublin Soc. 24 
319 (1948). 

442 Variolaric Acid (Ochrolechaic Acid, Parellic Acid), C16H10O7, 
colorless crystals, m.p. 296°. 




OH 



^D. H. R. Barton and T. Cohen, Festschrift Arthur Stoll, 111 
(1957). 

** Holger Erdtman and Carl Axel Wachmeister, ibid., 144 (1957). 

" Yasuhlko Asahina and Shoji Shibata, "Chemistry of Lichen Sub- 
stances," Japan Society for the Promotion of Science, Tokyo, 1954. 
(In English) 



215 Depsides and Depsidones 

Lecanora parella Ach. 

The yield was about 1 ^^ . Mannitol also was present. 
D. Murphy, J. Keane and T. J. Nolan, Sci. Proc. Roy. Dublin 
Soc. 23 71 (1943). 

443 Lecanoric Acid (Glabratic Acid), C16H14O7, colorless needles, 
m.p. 175°. 



O 

CH 



C— O OH 




HO OH ! COOH 

CH3 

Parmelia tinctorum Despr., P. horreri Turm., P. scortea 
Ach. and P. latissima Fee. 

Emil Fischer and Hermann O. L. Fischer, Ber. 46 1138 

(1913). (Synthesis) 

444 Diploschistesic Acid, C^Hi^Os, colorless leaflets, m.p. 174°. 

CH3 




CH. ^°°" 



Diploschistes scruposus (L.) and D. bryophilus (Ehrh.) 
Lecanoric acid was isolated from the same source. 
Yasuhiko Asahina and Masaichi Yasue, Ber. 69B 2327 
(1936). (Synthesis) 

445 Vicanicin, C17H14O5CI2, colorless needles, m.p. 248-250°. 

O 

CH, 
CI 




Teloschistes flavicans 

A yield of about 1% of the dry lichen weight was ob- 
tained. 



Pfizer Handbook of Microbial Metabolites 



216 



S. Neelakantan, T. R. Seshadri and S. S. Subramanian, 
Tetrahedron Letters No. 9, pp. 1-4 (1959). 

446 Evemic Acid, C17H16O7, colorless prisms, m.p. 169°. 

CH3 



CH3O 




COOH 



Evernia prunastri L., Ramalina pollinaria Wests., Us- 
nea jesoensis Asahina 

Fukuziro Fuzikawa and Kumao Ishiguro, /. Pharm. Soc. 
Japan 56 837 (in German, 149) (1936). (Synthesis) 

447 Norstictic Acid, CigHigOg, nearly colorless needles, m.p. 283° 
(dec). 




COO 



CH3 



CHO O 



OH 



CO 
CH— O 
OH 



Lobaria pulmonaria Hoffm., Parmelia acetabulum 
Duby., Usnea japonica, Wain., etc. 

Yasuhiko Asahina and Masaichi Yanagita, Ber. 67B 799 
(1934). " 

448 Salazinic Acid (Saxatilic Acid), CigHioOio, colorless needles, 
m.p. 260° (dec. from 240°). 

ru CH2OH 

^"^ COO I OH 




"° ino^o/ 



xo 

CH — O 

I 
OH 



217 Depsides and Depsidones 

Parmelia cetrata Ach., P. conspersa Ach., P. marmariza 
Nyl., P. saxatilis Ach., P. abyssinica Kremp. 

Yasuhiko Asahina and Juntaro Asano, Ber. 66B 689, 893, 
1215 (1933). 

449 Gangaleoidin, C1SH14O7CI2, colorless needles, m.p. 213°. 

^"^ OCH3 



COOCH3 



Lecanora gangaleoides Nyl. 

V. E. Davidson, J. Keane and T. J. Nolan, Sci. Proc. Roy. 
Dublin Soc. 23 143 (1943). (Structure) 

450 Psoromic Acid (Sulcatic Acid, Parelllc Acid), C18H14O8, colorless 
needles, m.p. 265°. 



COO 






CHO '^ COOH 

Psoroma crassum Korber, Alectoria zopfii Asahina, etc. 
Syozi Shibata, /. Pharm.. Soc. Japan 59 323 (in German, 
111) (1939). (Synthesis) 

451 Protocetraric Acid (Capraric Acid, Ramalinic Acid), C18H14O9, 
colorless fine needles, m.p. 250° (dec. from 220°). 

CH3 ^^^ CH2OH 
COO I OH 




CH3 ^°°" 

Parmelia caperata, Ramalina farinacea, etc. 

Yasuhiko Asahina and Yaichiru Tanase, Ber. 66B 700 
(1933). 

Yasuhiko Asahina and Juntaro Asano, ibid. 66B 893, 1215 
(1933). 



Pfizer Handbook of Microbial Metabolites 218 

452 Barbatolic Acid, C18H14O10, colorless crystals, m.p. 206° (dec.) 
(s. 190°). 

COOCH2 I OH 





OH ^"° 

Usnea barbata, Alectoria implexa (Hoffm ) Nyl. f. fus- 
cidula Am. 

Eero E. Suominen, Suomen Kemistileht; 12B 26 (1939). 

453 Pannarin, CisHi.PeCl, colorless prisms, m.p. 216°. 

Q ^"' 




CH,0 i.. \n/ 1„ CHO 
On 

Pannaria lanuginosa Korb., P. fulvescens Nyl., P. lurida 
Nyl. 

Itiro Yosioka, /. Pharm. Soc. Japan 61 332 (1941). 

454 Obtusatic Acid (Ramalic Acid), C^gHisO^, colorless needles, 
m.p. 208° (dec). 




CH3O 1.. OH 1^ COOH 

CHs 

Ramalina pollineria Ach. other Ramalina species 
Evemic acid, usnic acid and sometimes sekikaic acid 
were isolated from the same sources. 

Fukuziro Fuzikawa, J. Pharm. Soc. Japan 56 237 (in Ger- 
man, 25) (1936). (Synthesis) 



219 



Depsides and Depsidones 



455 Stictic Acid (Stictaic Acid, Pseudopsoromic Acid, Scopularic 
Acid), CiaHi^Ofl, colorless microcrystals, m.p. 268° (dec). 




Lobaria pulmonaria Hoffm., L. oregana Miill. Arg., 
Stereocaiilon nabewariense Zahlb., etc. 

Yasuhiko Asahina and Masai ti Yanagita, Ber. 67B 1965 
(1934). 

456 Nornidulin (Ustin), Ci^Hi-O^Cla, hexagonal plates or prisms, 
m.p. 185°. 




Aspergillus nidulans, NRRL No. 2006 

A little succinic acid was isolated from the same cul- 
ture. 

F. M. Dean, John C. Roberts and Alexander Robertson, /. 
Chem. Soc, 1432 (1954). 

457 Dechloronornidulin (Ustin II), CigHieOjCL, needles, m.p. 212- 
214°. 

Partial structure: 




Pfizer Handbook of Microbial Metabolites 220 

Aspergillus nidulans NRRL No. 2006 
F. M. Dean, A. D. T. Erni and Alexander Robertson, /. 
Chem. Soc, 3545 (1956). 

458 Thamnolic Acid, CigHigOn, pale yellow crystals, m.p. 223° 
(dec). 

^"' COO. ^"^ COOH 





CH3O I OH HO 

COOH 

Thamnolia vermicularis (Sw.) Schaer., Cladonia poly- 
dactyla Flk., CI. digitata, other Cladonia, Parmeliopsis 
and Pertusaria spp. 

Yasuhiko Asahlna and Michio Hiraiwa, Ber. 69B 330 
(1936). 

Idem., ibid. 72 1402 (1939). 

459 Chloroatranorin, CigHi^OsCl, colorless crystals, m.p. 208°. 

CH3 




Parmelia furfuracea Ach., P. physodes Ach., Evernia 
prunastri, etc., wide occurrence 

Georg Roller and Karl Popl, Monatsh. 64 106 (1934). 
Idem. f ibid. 64 126 (1934). 

460 Atranorin (Atranoric Acid, Usnarin, ParmeHn), CigHigOg, color- 
less prisms, m.p. 196°. 

^"' coo 





Atranorin occurs in about 90 different lichens. 
d-Usnic acid also often is present. 



221 Depsides and Depsidones 

Alexander St. Pfau, Helv. Chim. Acta 9 650 (1926). 
461 Baeomycesic Acid, CigHigOs, colorless crystals, m.p. 233°. 

CHa 




Baeomyces roseus Pers., JB. fungoides Ach., Thamnolia 
subverniicularis Asahina 

Squamatic acid also was present in some cases. 
Georg KoUer and Walter Maass, Monatsh. 66 57 (1935). 

462 Squamatic Acid (Sphaerophoric Acid), CigHjgOg, colorless crys- 
tals, m.p. 228° (dec). 




CHaO 



Cladonia bellidifiora var. coccocephala Ach., CI. squa- 
mosa Hoffm., CI. uncialis (L. ) Web., Thamnolia sub- 
vermiciilaris Asahina 

A little ^usnic acid was present also. 

Yasuhiko Asahina and Yoshio Sakurai, Ber. 70B 64 (1937). 
( Synthesis ) 



463 Hypothamnolic Acid, CigHigOjo, colorless needles, m.p. 217.5'^ 

COOH 



CHs /COO\ 9*^3 





/ 
CH3O ' ^^OH HO 
COOH 

Cladonia pseudostellata Asahina 

The yield was about 0.1%. Usnic acid was present 
also. 

Yasuhiko Asahina, Masaru Aoki and Fukuziro Fuzikawa, 
Ber. 74B 824 (1941). 



Pfizer Handbook of Microbial Metabolites 222 

464 Barbatic Acid (Rhizoic Acid, Coccellic Acid), C19H20O7, colorless 
prisms, m.p. 187° (dec.)- 




CH3O I OH I COOH 

Cladonia fioerkeana Sommerf., CI. bacillaris Nyl., CI. 
macilenta (Hoff. ) Flk., CI. coccifera (L. ), CI. amauro- 
craea (Fl.) Schaer., Rhizocarpon geographicum (L.), Us- 
nea longissima Ach. 

Usnic acid also was present. 

Fukuziro Fuzikawa, /. Pharm. Soc. Japan 56 237 (in Ger- 
man, 25) (1936). (Synthesis) 

465 Physodalic Acid (Monoacetylprotocetraric Acid), CooHjeOio, col- 
orless plates, m.p. 260° (dec. from 230°). 

CH3 ^^^ CH.OCOCH3 
COO I OH 




HO I \r,/ i COOH 

CHO O 

Parmelia physodes Ach., P. hypotrypella Asahina 
Wilhelm Zopf, Ann. 295 287 (1897). 
Idem., ibid. 300 350 (1898). 

466 Nidulin, CooHi^O.-.CL^, colorless crystals, m.p. 180°. 

O 

... II 

CI T c— o I 0CH3 



CsH, 



Aspergillus nidulans NRRL, No. 2006 

The yield was about 6 g. from 126 g. of dry mycelium; 
a little mannitol also was found. 

F. M. Dean, John C. Roberts and Alexander Robertson, J. 
Chem. Soc, 1432 (1954). 




223 Depsides and Depsidones 

467 Diffractaic Acid (Dirhizonic Acid), C00H00O7, colorless needles, 
m.p. 189°. 




CH3O \ OCH3 i COOH 
CH3 

Usnea diffracta Wain., Usnea longissima Ach., Alecto- 
ria ochroleuca Mass. 

The yield was 3.6%. 

Yasuhiko Asahina and Fukuziro Fuzikawa, Ber. 65B 583 
(1932). (Synthesis) 

468 Erythrin, CmuHooOj,, colorless needles, m.p. 148°, [ocW^ +10.63°. 
?"^ COO OH 




HO OH 1 COOCH2— CH— CH— CH2 

CHii I I I 

OH OH OH 

Roccella montagnei Bel. and R. fuciformis DC 
This is an erythritol ester of lecanoric acid. The yield 
was about 5% of the weight of the lichen. Free erythritol 
and rocellic acid were isolated from the same source. 

Yosio Sakurai, 7. Pharni. Soc. Japan 61 108 (in German, 
45) (1941). 

469 Divaricatic Acid, C21H04O7, colorless needles, m.p. 137°. 

CH3CH2H,C 




CH3O OH I COOH 

CH2CH2CH3 

Evernia divaricata L., E. mesomorpha f. esorediosa 
Miill., Arg. 

The yield from E. mesomorpha was recorded as 2.5% 
of the lichen weight. Usnic acid was isolated from the 
same source. 



Pfizer Handbook of Microbial Metabolites 224 

Yasuhiko Asahina and Michio Hiraiwa, Ber. 70B 1826 
(1937). (Synthesis) 

470 Fumarprotocetraric Acid, C22H16O12, colorless needles, m.p. 250— 
260° (dec. from 230°). 

CH3 , CH2OOC— CH=CH— COOH 

COO I OH 




Cetraria islandica Ach., Cladonia rangiferina (L. ) 
Web., CI. sylvatica (L.) Hoffm. 

Yasuhiko Asahina and Yaichiro Tanase, Ber. 67B 766 
(1934). 

471 Sekikaic Acid, C22H26O8, colorless needles, m.p. 147° (dec). 

CH3CH2CH2 .COO. OH ^^^,. 

\ I COOH 





CH3O OH CH3O CH2CH2CH3 

Ramalina geniculata Hook et Tayl., R. calicaris Rohl, 
and R. intermediella Wain. 

The yield was about 1 % . A little d-usnic acid also was 
present as well as ramalinolic acid. 

Yasuhiko Asahina and Masaichi Yasue, Ber. 68B 132 
(1935). (Synthesis) 

472 Sphaerophorin, C23H28O7, colorless crystals, m.p. 137°. 
CH3 




CH3O OH I COOH 

CHoCHoCH^CHoCHoCHiCHs 

Sphaerophorus fragilis Pers., S. coralloides Pers., S. mel- 
anocarpus 



225 Depsides and Depsidones 

Akira Hasimoto, /. Pharm. Soc. Japan 58 776 (in German, 
221) (1938). (Synthesis) 

473 Imbricaric Acid, C23H28O7, colorless needles, m.p. 125°. 
CH3CH2CH2CH2H2C 




CH3O OH I COOH 

CH2CH2CH3 

Parmelia perlata Ach., Cladonia impexa Harm., CI. 
evansi f. Abb., CI. pseudoevansi Asahina 

Yasuhiko Asahina and Itiro Yoshioka, Ber. 70B 1823 
(1937). (Synthesis) 

474 Ramalinolic Acid, CogHogOy, colorless crystals, m.p. 163°. 

CH,CH,CH, cOO^ OH ^^^^ 



CH3O OH HO CH2CH2CH2CH2CH3 

Ramalina intermediella Wain., R. calicaris Rohl, R. ge- 
niculata Hook et Tayl. and R. usneoides Mont. 

Yasuhiko Asahina and Tunaharu Kusaka, Ber. 69B 1896 
(1936). (Synthesis) 

475 Gyrophoric Acid, C24H20O10, colorless needles, m.p. 220°. 
^"^ COO OH ^"^ COOH 





HO OH 



Gyrophora esculenta Miyoshi, G. proboscidea L., Um- 
bilicaria pustulata L. Hoffm. Ochrolechia pallescens, Lo- 
baria pulmonaria var. meridionalis (Wain.) Zahlbr. 

Yasuhiko Asahina and Itiro Yasioka, Ber. 70B 200 (1937). 
(Synthesis) 



Pfizer Handbook of Microbial Metabolites 226 

476 Hiascic Acid, C24H20OH, colorless crystals, m.p. 190.5° (dec.)- 

CH3 

COOH 




Cetraria hiascens Th. Fr. 
Gyrophoric acid also was present. 

Yasuhiko Asahina and Tunaharu Kusaka, Bull. Chem. Soc. 
Japan 17 152 (in German) (1942). 

477 Anziaic Acid, C04H30O7, colorless, fine needles, m.p. 124° (dec). 
CHsCH-CHoCHoHoC 




COOH 

CH2CH2CH.2CH2CH3 

Anzia opuntiella Miill. Arg., A. gracilis, A. leucobatoides 
f. hypomelaena and Cetraria sanguinea 

Yasuhiko Asahina and Michio Hiraiwa, Ber. 70B 1826 
(1937). (Synthesis) 

478 Homosekikaic Acid (Nemoxynic Acid), C24H30O8, colorless 
prisms, m.p. 137.5°. 



CH3CH2CH2 .COO, 



COOH 



CH3O OH CH3O CH2CH2CH2CH2CH3 

Cladonia pityrea Flk. f. phyllophora Mudd, Cladonia 
nemoxyna (Ach.) Nyl. 

The yield was about 0.1%. A little fumarprotocetraric 
acid also was present. 

Yasuhiko Asahina and Tsunakaru Kusaka, Ber. 70B 1815 
(1937). (Synthesis) 



227 Depsides and Depsidones 

479 Umbilicaric Acid, Co-.HooO,,,, colorless crystals, m.p. 203° (dec.)- 
Synthetic sample m.p. 189°. 

COOH 




Gyrophora polyphylla (L.), G. deiista (L.) and G. vel- 
lea (L. ) 

Yasuhiko Asahina and Itiro Yosioka, Ber. 70B 200 (1937). 
(Synthesis) 

480 Lobaric Acid ( Stereocaulic Acid, Usnetic Acid), Co.-.HofiOs;, color- 
less needles, m.p. 192°. 

CH3CH0CH0CH0CO ^^^ 

I COO OH 




CH3O v,/ i ^°o" 



CHoCHoCHoCHaCHs 

Stereocaulon paschale Ach., S. exutum Nyl., etc. (wide 
occurrence ) 

Yasuhiko Asahina and Masaiti Yasue, Ber. 69B 643 (1936). 

481 Glomelliferic Acid, Co-.H^oOs, colorless prisms, m.p. 143°. 
CH3CH2CH2COH2C 




CH3O OH I XOOH 

CH2CH2CHCH2CH3 

Parmelia glomellifera Nyl. 
W. Zopf, Ann. 297 303 (1897), 313 341 (1900). 
Yasuhiko Asahina and Hisasi Nogami, Ber. 70B 1498 
(1937). 

K. Mlnami, J. Pharm. Soc. Japan 64 315 (1944). 



Pfizer Handbook of Microbial Metabolites 228 

482 Perlatolic Acid, C25H32O7, colorless needles, m.p. 108°. 
CH3CH2Cn2Cn2n2C 




COOH 
CH2CH2CH2CH2CH3 

Parmelia perlata Ach., Cladonia impexa Harm., 
CI. evansi f. Abb., CI. pseudoevansi Asahina 

Yasuhiko Asahina and Itiro Yoshioka, Ber. 70B 1823 

(1937). (Synthesis) 

483 Boninic Acid, C25H32O8, colorless plates, m.p. 134.5°. 

COO 
CH3CH2CH2 / \ OH ^^^^ 




CH3O OCH3 CH3O CH2CH2CH2CH2CH3 

Ramalina boninensis Asahina 

The yield was about 0.5%, and a little d-usnic acid was 
present. 

Yasuhiko Asahina and Tsunaharu Kusaka, Ber. 70B 1815 
(1937). (Synthesis) 

484 Tenuiorin, C26H24O10, colorless crystals, m.p. 238° (dec. ) s. 180°. 

O 

^"^ COOCH3 




CH3O OH 



Lobaria pulmonaria Hoffm. f. tenuior Hue. 
Mannitol also was present. 

Yasuhiko Asahina and Masaiti Yanagita, Ber. 66B 1910 
(1933). 



229 Depsides and Depsidones 

485 Physodic Acid (Farinacic Acid), CoeHagOg, colorless prisms, m.p. 
205° (dec). 

CH3CH2CH,CHoCH2COCH2 ^^^ 

COO OH 




COOH 
CH2CH2CH2CH2CH3 

Parmelia physodes Ach., P. furfuracea Ach, 
A yield of 5% was reported. 

Yasuhiko Asahina and Hirasi Nogami, Ber. 67B 805 
(1934). 

Idem., ibid. 68B 77, 1500 (1935). 

486 Olivetoric Acid, CogHgoOg, colorless crystals, m.p. 151°. 

CH3CH2CH2CH2CH2COH2C 

COO OH 




COOH 
CH2CH2CH2CH2CH3 

Parmelia olivetorum Nyl., Comicularia pseudosatoana 
Asahina and C. divergens Ach. 

Yasuhiko Asahina and Fukuziro Fuzikawa, Ber. 67B 163 

(1934). 

487 Alectoronic Acid, C28H32O9, colorless prisms, m.p. 193°. 
CH3CH2CH2CH2CH2COCH2 




COOH 
CH2COCH2CH2CH2CH2CH3 

Alectoria japonica Tuck., A. sarmentosa Ach., Cetraria 
pseudocomplicata Asahina, Nephromopsis cilialis Hue. 

Yasuhiko Asahina, Yoshinari Kanaoka and Fukuziro Fu- 
zikawa, Ber. 66B 649 (1933). 



Pfizer Handbook of Microbial Metabolites 230 

488 a-Collatolic Acid (Lecanorolic Acid, Lecanoral), C29H34O9 color- 
less needles, m.p. 124°. 

CH3CH0CH.2CH2CH0COCH2 ^^^ ^.. 

COO OH 




CH3O \r,^ I COOH 

CH2COCH2CH2CH2CH2CH3 

Cetraria collata Miill. Arg., Lecanora atra (Hudson) 
Ach., L. grumosa (Pers.) Rohl. 

Yasuhiko Asahina, Yoshinari Kanaoka and Fukuziro Fu- 
zikawa, Ber. 66B 649 (1933). 

489 Microphyllic Acid, C29H36O9, colorless needles, m.p. 116°. 
CH3CH2CH2CH2CH2COH2C 




CHsO^ ^OH I XOOH 

CH2COCH2CH2CH2CH2CH3 

Cetraria japonica Zahlbr. 

Some chloroatranorin was isolated from the same ex- 
tract. The yield of microphyllic acid was about 4^^ of 
the lichen weight. 

Yasuhiko Asahina and Fukuziro Fuzikawa, Ber. 68B 2022 
(1935). 



12 



Quinones and Related Compounds 



Quinones occur widely in nature, and this topic has been re- 
viewed.^- -■ ■"' Even allowing for their conspicuousness due to 
color, solubility characteristics and (often) quantity, it seems 
that they are broadly distributed among plants, and fungi are 
no exceptions. 

Anthraquinones, in particular, have been isolated frequently 
from fungus cultures. Some 80 anthraquinones and related 
substances of known structure were listed by W. Karrer'^ as hav- 
ing been reported from plant sources in general. About half of 
this number have been isolated and characterized from fungi 
and lichens. Since no anthraquinones have been reported from 
algae, it may be that those present in lichens are formed pri- 
marily by the fungus component. There is some evidence, how- 
ever, that in lichens both partners are required for the biosyn- 
thesis of depsides and depsidones.^ 

In fungi anthraquinones occur mainly in the mycelium, often 
as mixtures of closely related materials. It is likely, for this 
reason, that some of the quinones reported in the early litera- 
ture were impure. 

The frequent identification of anthraquinone pigments in 
molds has caused some speculation on their function. Argu- 
ments in favor of a biological function have been summarized 

^ S. Shibata, Kagaku (Science) 26 391-396 (1956). 

~ R. H. Thomson, "Naturally Occurring Quinones," Academic 
Press, New York, 1957, 302 pp. 

' W. Karrer, "Konstitution und Vorkommen der Organischen 
Pflanzenstoffe," Birkhauser Verlag, Basel, 1958. 

* Dieter Hess, Zeitschr. Naturforsch. 14b 345 (1959). 



Pfizer Handbook of Microbial Metabolites 232 

as follows : ^ ( 1 ) Some pigment complexes are produced in 
large quantities, up to 30 percent of the total dry weight of the 
mycelium. (2) In many cases, the maximal pigment content 
is reached while usable carbohydrate is still present. If har- 
vesting is delayed, pigment disappears as autolysis sets in. 
(3) The same pigment often is present in different genera or 
families, suggesting solution of a metabolic problem in the 
same way. (4) Reduction products such as anthranols, an- 
thrones and quinhydrones sometimes are present together with 
the parent quinone, perhaps indicating a hydrogen or electron 
transport function. (5) Several mold pigments are antibiotic 
toward other fungi and bacteria. 

On the other hand, it has been pointed out" that, in fungi, 
induced mutations leading to full blocking of the production of 
acetate-derived aromatic compounds such as anthraquinones do 
not seem to affect the vitality of the organism. The antifunc- 
tionalists believe that anthraquinones and perhaps some other 
mold metabolites are merely waste or storage products due to 
an overflow of acetate metabolism. If some of these products 
happen to inhibit competitors, they facilitate species survival. 

A similar concept of the significance of such mold metabolites 
has been mentioned by Dalgliesh.^ He proposed that enzyme 
systems unable to deal with substrate because it is in large ex- 
cess, or for some other reason, might convert it to anthraqui- 
nones and other substances, which are eliminated, then, in a 
kind of "detoxication" disposal mechanism. 

An enzyme chemist, F. F. Nord, has suggested- that many of 
the metabolites produced in yields exceeding functional re- 
quirements, or for which there is no function, accumulate be- 
cause some of- the enzyme systems involved in the oxidative 
sequences become saturated with respect to their substrates. 
They are thus, in his opinion, probably products of anaerobic 

^ G. Smith, Congr. intern, botan. Paris, Rapps. et communs., 8 
Sec. 83-89 (1954). 

^' Gosta Ehrensvard, "Developments in Aromatic Chemistry," Spe- 
cial Publication No. 12, English Chemical Society, London, 1958, 
p. 29. 

7 C. E. Dalgliesh, ibid., p. 14. 

^ F. F. Nord and D. D. Clarke, Arch. Biochem. and Biophys. 59 
285 (1955). 



233 



Quinones 



metabolism, and arise in a manner analogous to the accumula- 
tion of citric acid, which is induced under the same conditions.^ 

There is no convincing experimental evidence that anthra- 
quinones are important in electron transport. 

It has been suggested^" that anthraquinones are acetate-de- 
rived, and there is some experimental confirmation.^^' ^^' * 

This proof was obtained by growing the mold in the presence 
of C'*-labeled acetate, isolating the metabolite, which incorpo- 
rated the label to some degree, then degrading the molecule by 
ingenious chemical methods to determine the sites of labeling. 

Although an acetate origin is indicated, the detailed natures 
of the intermediates in the biosynthetic mechanism are still 
unknown. Intermediates such as orselHnic acid,^^ dihydroxy- 
phthalic acid,^* and 6-methylsalicylic acid^^ (all known mold 
metabolites) have been proposed, e.g.: 




COOH 



COOH 



3,5-Dihydroxy- 
phthalic Acid 




COOH 



6-Methylsali 
cylic Acid 



Birch prefers to think in terms of an intermediate formally 
resembling a polyketomethylene chain, which can be modified 
in various ways on an enzyme surface to yield related metabo- 
lites. This concept is supported by the occasional discovery of 
related metabolites in the same culture or plant. For example, 
the co-occurring anthraquinone and phenanthrenequinone 

9 H. A. Krebs, Biochem. J. 31 2095 (1937). 

"A. J. Birch and F. W. Donovan, Austral. J. Chem. 6 360 (1953). 

" Sten Gatenbeck, Acta Chem. Scand. 12 1211 (1958). 

12 A. J. Birch, A. J. Ryan and Herchel Smith, /. Chem. Soc, 4473 
(1958). 

* Also see addendum for later work. 

1^ K. Aghoramurthy and T. R. Seshadrl, J. Sci. Ind. Res. (India) 
13A 114 (1959). 

"E. L. Tatum, Ann. Rev. Biochem. 13 667 (1944). 

15 Harold Raistrick, Acta Chem. Fenn. lOA 237 (1950). 



Pfizer Handbook of Microbial Metabolites 



234 



shown below could be envisaged as derivatives of a common 
precursor chain, which is laid down upon the enzyme surface in 
different patterns before cyclization.^^ 

O 




It is likely that the dianthraquinones are formed by oxidative 
phenoUc radical coupling, e.g.:* 




HO O OH 
Skyrin 



^'' A. J. Birch, private communication. 

* See addendum for evidence to the contrary. 



235 



Quinones 



Other metabolites, such as actinorhodhi and the perylene- 
quinones may be formed similarly. 

Structures such as moUisin and javanicin seem to indicate an 
acetate derivation for naphthoquinones. 




CH3O 



HO 



O 
CHo— C— CH3 




OH O 



CH, 
Mollisin 



Javanicin 



The suggestion has been made^' that the terphenylquinones 
might be formed by autocondensation of a phenylpyruvic acid 
type of molecule in the following sense: 



o— 



c=o 

O— R 



R— O 
C 

o c 



CH 



,-r\ 



/ 



/ 



o \= 



Phenylpyruvcte 



OH 



HO 



o \=/ 

Polyporic Acid 

Similarly p-hydroxyphenylpyruvate would form atromentin. 
Polyporic acid might be transformed by oxidation to pulvinic 

^'G. Read and L. Vining, Chem. and Ind., 1546 (1959). 



Pfizer Handbook of Microbial Metabolites 236 

acid, and by further hydroxylations to leucomelone or other 
terphenylquinones . 

O 

\ 

c o 



c c 

\/\ 

c c- 




o c 

\ 

o 

Pulvinic Acid Leucomelone 

If this suggestion can be confirmed experimentally, it will re- 
late this type of benzoquinone metabolite to the shikimic acid 
route of biogenesis. 

The biosynthesis of a benzoquinone, aurantiogliocladin, has 
been studied, by using C"-labeled formate and acetate. ^^ The 
results demonstrated formation from 4 moles of acetate with de- 
carboxylation, C-methylation, post-oxidation in the aromatic 



CH3^ 
' Q 



% O 

C— CH3 CH3O II CHa 




O -^ 

CH3— ic=o] 

Acetate Aurantiogliocladin 

ring and 0-methylation of the phenolic hydroxyl groups. 
6-Methylsalicylic acid appears to be an intermediate.^^ 
Aurantiogliocladin, isolated from a gliocladium specimen, re- 
sembles the coenzymes Q. These substances occur in the cell 
mitochondria of a wide variety of organisms. They are benzo- 
quinones substituted similarly to aurantiogliocladin, but with 

^^ A. J. Birch, R. I. Fryer and Herchel Smith, Proc. Chem. Soc, 
343 (1958). 

19 Private communication from Herchel Smith. 



237 



Quinones 



additional polyisoprenoid side-chains. There is a marked re- 



CH3O 



CH3O 




CH3 

I 
(CH2— CH=C— CH2)nH 

Vitamin Kq 

semblance to the previously discovered vitamins K. The follow- 
ing substances have been isolated, purified, and the structures 
determined : 



TABLE I 





Numbers of 










Origin 


side-chain 
isoprene 
units (n) 


Number of 
carbon 
atoms 


Melting 
point (°C) 


Designation 


Refer- 
ences 


Saccharomyces cere- 












yisiae 


6 


39 


16° 


Coenzyme Qe 


20 


Torula utilis 


7 


44 


30.5° 


Coenzyme Q7 


20,21 


Azotobacfer vine- 












landii 


8 


49 


37° 


Coenzyme Qg 


20,21 


Torula utilis 


9 


54 


45.2° 


Coenzyme Q9 


20,21 


Beef heart 


10 


59 


48° 


Coenzyme Qio 


21,22 



A survey was made, by using methods sometimes short of iso- 
lation and purification (paper chromatographic comparisons, 
spectra, etc. ) of the occurrence of coenzyme Q and of vitamin K 
in a wide variety of biological types. ^^ Many bacteria contain 
coenzyme Q. The mycobacteria and streptomycetes seem to 
contain vitamin K instead. Escherichia coli and chromatium 
species contain both. Obligate anaerobes such as the Clostridia 

20 R. L. Lester, F. L. Crane and Y. Hatefi, J. Am. Chem. Soc. 80 
4751 (1958). 

-1 R. L. Lester and F. L. Crane, Biochim. et Biophys. Acta 32 492 
(1959). 

2- F. L. Crane, Y. Hatefi, R. L. Lester and Carl Widmer, Biochim. 
et Biophys. Acta 25 220 (1957); idem., ibid. 32 73 (1959). 

23 R. L. Lester and F. L. Crane, /. Biol. Chem. 234 2169 (1959). 



Pfizer Handbook of Microbial Metabolites 



238 



contain neither, and facultative anaerobes such as Saccharomy- 
ces cerevisiae and E. coli contain neither when grown anaero- 
bically. A chart of microbial occurrence was published: 

TABLE II 



Organism 


Coenzyme Q 


Vitamin K 


Saccharomyces cerevisiae (anaerobic) 


- 


- 


Saccbaromyces cerevisiae (aerobic) 


Qe 




Socchoromyces cavalier! 


Qe 




Saccharomycei fragilis 


Qe 




Neurospora crassa 


Qio 




Mucor corymbifer 


Qs) 




Sfrepfomyces griseus 




+ 


Mycobacferium smegmafis 




+ 


Mycobacterium tuberculosis 




+ 


Bacillus mesentericus 




+ 


Escherichia coli 


Qh 




Chromatium spp. 


Q7 




Rhodospii ilium rubrum 


Q9 




Pseudomonas fluorescens 


Qs 




Hydrogenomonas sp. 


Qs 





Basidiomycetes contain neither coenzyme Q nor vitamin K, 
but produce another quinone which seems to have the same 
function in this family. It has been extracted and purified to 
some extent and called basidioquinone. 

A comparison of all the animal, plant and microorganism 
sources indicated that, in general, lower organisms contain 
lower homologues of coenzyme Q. 

Evidence has been obtained for the coenzyme function of the 
Q (and K) quihones: ( 1 ) Extraction from mitochondria destroys 
enzymatic activity, which is restored by restoration of the coen- 
zymes. (2) Inhibitors of electron transport, such as the anti- 
biotic, antimycin A, affect the oxidation state of the quinones in 
a predictable manner. (3) The rate of oxidation or reduction 
in mitochondria is what might be anticipated for participation 
in electron transport. The pattern of occurrence in aerobic and 
anaerobic microorganisms also is suggestive. 

The general structure of the electron transport system in cell 
mitochondria in the light of the new discoveries has been re- 
viewed.^* 

Apparently coenzyme Q is formed by a combination of the 

-^ D. E. Green and R. L. Lester, Federation Proc. 18 987-1000 
(1959). 



239 Quinones 

simple acetate and terpenoid biosynthetic routes. Mevalonic 
acid was incorporated into the molecule by rats (especially vita- 
min A-deficient rats) and by rat liver, while 2,3-dimethoxy-5- 
methyl-l,4-benzoquinone and D,L-tocopherol were not.'-'^ This 
contrasts with evidence that 2-methyl-l,4-naphthoquinone is 
used as a precursor of vitamin K by rats.'-" Evidently, no experi- 
mental work has been published on biosynthesis in microor- 
ganisms. 

a. BENZOQUINONES 

490 Tetrahydroxybenzoquinone, C,,H40(j, bluish black plates, no 
melting point. 

O 

HO II OH 



HO 11 OH 
O 

Pseudomonas beiierinckii Hof grown on salted beans. 
The substrate is meso-inositol, which probably is a normal 
constituent of beans. 

T. Hof, Rec. Trav. Botan. Neerland. 32 92 (1935). (Isola- 
tion) 

A. J. Kluyver, T. Hof and A. G. J. Boezaardt, Enzymologia 
7 257 (1939). (Structure) 

Paul W. Preisler and Louis Berger, /. Am. Chem. Soc. 64 
67 (1942). 

491 Gentisylquinone, CjHcOs, yellow needles, m.p. 76°. 

O 

CHoOH 




O 

PenicilliuTn patulum Bainier probably produces a little 
of this quinone under certain conditions, although it may 
be an artifact, since larger quantities of the corresponding 
hydroquinone are produced. It has been isolated as a 
deep violet colored complex, m.p. 86-89°, with the hydro- 
quinone. 

-' U. Gloor and O. Wiss, Arch. Biochem. and Biophys. 83 216 
(1959). 

-«C. Martius and H. O. Esser, Biochem. Z. 331 1 (1958). 



Pfizer Handbook of Microbial Metabolites 240 

B. G. Engel and W. Brzeski, Helv. Chim. Acta 30 1472 

(1947). 

492 Terreic Acid, C7H6O4, pale yellow, large, glistening plates, m.p. 
127-127.5°, [aln" -28.6° (c 1 in 50% methanol-benzene). 
Sublimes. 




Aspergillus terreus grown in a glucose and corn-steep 
liquor-cottonseed meal medium. 

H. M. Florey, E. Chain, N. G. Heatley, M. A. Jennings, A. G. 
Sanders, E. P. Abraham and M. E. Florey, "Antibiotics," Ox- 
ford University Press, London, 1949 Vol. I p. 388. 

Murray A. Kaplan, Irving R. Harper and Bernard Heine- 
mann. Antibiotics and Chemotherapy 4 746 (1954). Yield 
138 g. from 200 liters. 

J. Sheehan, W. Lawson and R. Gaul, J. Am. Chem. Soc. 
80 5536-5538 (1958). (Structure) 

493 4-Methoxytoluquinone (Coprinin), CgHgOg, yellow spangles, 
m.p. 175°. 

CH3 ? 



OCH3 



Coprinus similis B. and Br., Lentinus degener Kalchbr. 
grown on a Czapek-Dox medium, containing glucose and 
corn-steep solids. 

Marjorie Anchel, Annette Hervey, Frederick Kavanagh, 
Jerome Polatnick and WUliam J. Robbins, Proc. Nat. Acad. 
Sci. U. S. 34 498 (1948). (Isolation) 

R. B. Woodward, Franz Sondheimer, David Taub, Karl 
Heusler and W. M. McLamore, /. Am. Chem. Soc. 74 4234 
(1952). (Synthesis) 



241 Benzoquinones 

494 2,5-Diniethoxybenzoquinone, C8H8O4, yellow prisms, in.p. 250° 
(dec). 




CH3O 



Polyporus fumosus (Pers.) Fries grown on an artificial 
medium including glucose and corn-steep liquor. 

Yield: 0.1 g. from 2 liters of culture broth. 

J. D. Bu'Lock, J. Chem. Soc, 575 (1955). (Isolation) 

E. Knoevenagel and C. Biickel, Ber. 34 3993 (1901). (Syn- 
thesis) 



495 Fumigatin, C8H8O4, maroon needles, m.p. 116' 



CH3 ? OH 







n OCH3 



496 Fumigatin Hydroquinone is produced as well, the ratio of the 
two compounds varying with the age of the culture. 




OH °<=* 



Aspergillus fumigatus Fres. grown on a Raulin-Thom 
medium. 

Winston Kennay Anslow and Harold Raistrick, Biochem. J. 
32 687 (1938). (Isolation) 

W. K. Anslow, J. N. Ashley and H. Raistrick, /. Chem. Soc, 
439 (1938). (Synthesis) 



Pfizer Handbook of Microbial Metabolites 242 

497 Spinulosin, CgHgOg, purple-black plates, m.p. 203°. 

CH3 ? OH 



HO I OCH3 



First isolated from three strains of Penicillium spinu- 
losiim Thorn grown on a modified Czapek-Dox-glucose 
medium. Later isolated from two out of seven strains of 
Aspergillus fumigatus examined. Spinulosin as well as 
an orange pigment, m.p. 184-185°, with antibiotic prop- 
erties resembling those of fumigatin, also has been iso- 
lated from an unidentified Penicillium (perhaps Penicil- 
lium spinulosumy Penicillium cinerascens Biourge is 
another producer. 

J. H. Birkinshaw and H. Raistrick, Trans. Roy. Soc. (Lon- 
don) B220 245 (1931). 

Winston K. Anslow and Harold Raistrick, Biochem. J. 32 
687, 2288 (1938). (Isolation) 

A. Bracken and H. Raistrick, ibid. 41 569 (1947). 

Keichiro Hoshishima, Tohuku J. Exptl. Med. 52 273 (1950). 

Winston K. Anslow and Harold Raistrick, Biochem. J. 32 
803 (1938). (Synthesis) 

498 Aurantiogliocladin, CioHjoOj, orange plates, m.p. 62.5°. 



CH3 II OCH3 



CH3 I OCH3 



The corresponding quinhydrone, a dark red compound 
called rubrogliocladin, occurs together with aurantioglio- 
cladin. 

A Gliocladium specimen, probably G. roseum Bainier 
produces these substances as well as : 



243 



Benzoquinones 



499 Gliorosein, C,oHj404, colorless crystals, m.p. 48' 



CHa 



CH3 




OCH3 



OCH3 



CH3 



CH3 




OCH3 



OCH3 



P. W. Brian, P. J. Curtis, S. R. Rowland, E. G. Jeffreys and 
H. Raudnitz, Experientia 7 266 (1951). (Isolation) 
E. B. Vischer, /. Chem. Soc, 815 (1953). (Structure) 
Wilson Baker, J. F. W. McOmie and D. Miles, ibid., 820 
(1953). (Synthesis) 

500 Phoenicin, C,4H,„0,., yellow-brown tablets, m.p. 231°. 

O HO O 




Penicillium phoeniceum van Beyma, P. ruhrum O. Stoll. 
Theodore Posternak, Hans W. Ruelius and Jacques Tcher- 
niak, Helv. Chivi. Acta 26 2031 (1943). (Synthesis) 

501 Oosporein (Chaetomidin), Ci^HmOs, bronze plates, m.p. 260- 

275°. 




Oospora colorans van Beyma, Chaetomium aureum 
Chivers, Verticillium psalliotae, Acremonium sp. 

F. Kogl and G. C. Van Wessem, Rec. trav. chim. 63 5 
(1944). (Isolation) 

F. M. Dean, A. M. Osman and Alexander Robertson, 
/. Chem. Soc, 11 (1955). 

G. Lloyd, Alexander Robertson, G. B. Sankey and W. B. 
Whalley, ibid., 2163 (1955). 



Pfizer Handbook of Microbial Metabolites 244 

502 Isooosporein,* C14H10O8, purple crystals, no m.p., subl. 220- 
250°, dec. 250°. 

O 

HO-// A // 




Unclassified citric acid-forming fungus. 
Maximal yield 2.5 g. per liter. 

Nobuyo Shigematsu, ]. Inst. Polytech., Osaka City Univ. 
Ser. C 5 100 (1956). 

503 Volucrisporin, C18H12O4, red plates, m.p. >300°. 

HO 



o W 

OH 

Volucrispora aurantiaca 

Occasionally small quantities of the leuco derivative 
(hydroquinone) occur with the pigment. 

P. V. Divekar, G. Read and L. C. Vining, Chem. and Ind., 
731 (1959). 

504 Polyporic Acid, C18H12O4, bronze leaflets, m.p. 305-307° (dec). 

? OH 





/ 
HO " 
O 

Polyporus nidulans Fries, P. rutilans (Pers.) Fries, 
Peniophora fdamentosa (B. and C.) Burt, Sticta coronata 
Muell., S. colensoi Bab. 

Fritz Kogl, Ann. 465 243 (1928). 

J. Murray, J. Chem. Soc, 1345 (1952). 

* See addendum. 



245 Benzoquinones 

The air-dried fruiting body of P. rutilans contains 23%. 
It is not produced by the fungal mycelium in artificial 
culture. 

Robert L. Frank, George R. Clark and James N. Coker, 
J. Am. Chem. Soc. 72 1824 (1950). 

Polyporic acid is probably identical with the lichen pig- 
ment, orygmaeic acid, first described by Zopf. 

Wilhelm Zopf, Ann. 317 124 (1901). 

505 Atromentin, CjsHioOe, bronze leaflets, no m.p. 



HO A , „ 

OH 

Paxillus atromentosus (Batsch.) Fr. 

This basidiomycete often grows on old tree trunks and 
produces the pigment first in a leuco-form, which air- 
oxidizes to the colored form on the outer portions of the 
fruiting body and during isolation. The yield was about 
2% of the weight of the air-dried fruiting body. 

Fritz Kogl, Ann. 465 243 (1928). 

506 Leucomelone, CigHiaOj, brown leaflets, m.p. 320° (dec). 



HO HO II /\ 

Polyporus leucomelas Pers. ex Fr. 
Yield 3 g. per kilogram of fruiting body. 
Masuo Akagi, /. Pharm. Soc. Japan 62 129 (1942). Syn- 
thesis) 

507 Thelephoric Acid, C20H12O9, lustrous, nearly black prisms, no 
m.p. 



OH 
HO- 




Pfizer Handbook of Microbial Metabolites 



246 



Partial structure: 



HO 



OH 



2 CH2O2 
1 OH 



Thelephora palmata, other Thelephora spp., Lobaria 
retigera Trev., L. pulmonaria (L.) HofFm., Hydnum spp., 
Cantharelliis multiplex Underw., Polystictus versicolor 
(L.)Fr. 

Fritz Kogl, Hanni Erxleben and Ludwig Janecke, Ann. 482 
105 (1930). 

K. Aghoramurthy, K. G. Sarma and T. R. Seshadri, Tetra- 
hedron Letters No. 8 20 (1959). (Revised structure) 

508 Muscarufin, C25Hi609-H20, orange-red needles, m.p. 275.5°. 

COOH 
HO ? 



CH=CH— CH=CH— COOH 



COOH 



Amanita muscaria (Linn.) Fries 

This pigment causes the red color of the caps of this 
common poisonous toadstool (fly agaric), yet 500 kg. of 
the fungus yielded only 850 mg. of pure material. 

Fritz Kogl and Hanni Erxleben Ann. 479 11 (1930). 

509 Auriantiacin (Atromentin-3,6-dibenzoate), CgoHooOj-, dark red 
needles, m.p. 285-295°. 

O 

o— c- 




Hydnum aurantiacum Batsch. 



247 Benzoquinones 

Jarl Gripenberg, Acta Chem. Scand. 10 1111 (1956). 

510 Protoleucomelone, C32HosO,4, colorless crystals, m.p. 203-205°. 
Probable structure: 

CH3OCO ^^-^ 
CH3OCO. I / V-OCOCHs 
CH3COO \r^^/ \=/ 




CHaOCO-f y y ^OCOCH3 

OCOCH3 

Polyporus leiicomelas Pers. ex Fr. 

Yield 3-4 g. per kilogram of mushrooms. 

Masuo Akagi, /. Pharm. Soc. Japan 62 129 (1942). 

511 Metabolite of Hydnum aurantiarum, C^<iii^^)OlQ, colorless needles. 

m.p. 305-307°. 




(y^--" <r^o°" 



° i=o 



Hydnum aurantiacinn Batsch. 

Aurantiacin and thelephoric acid are produced by the 
same organism. 

Jarl Gripenberg, Acta Chem. Scand. 12 1411 (1958). 
Coenzymes Q (Mitoquinone, Ubiquinone, Qot.-,. SA). 

These compounds occur widely in the cell mitochondria 
of microorganisms and higher animals, where they play a 
part in the electron transport system. Variations in side- 
chain length occur as in the case of vitamin K. Com- 
pounds in which n = 6, 7, 8 and 9 have been isolated from 
microbial sources. The quinone moiety resembles auran- 
tiogliocladin. 



Pfizer Handbook of Microbial Metabolites 248 

General structure: 

O 
CH3O 




CH3O 



512 Coenzyme Qg, C39H58O4, m.p. 16°. 

Saccharomyces cerevisiae 

513 Coenzyme Q7, C44H66O4, orange crystals, m.p. 30.5°. 

Torula utilis 

514 Coenzyme Qg, C49H74O4, orange crystals, m.p. 37°. 

Azotobacter vinelandii 

515 Coenzyme Qg, C54H82O4, orange crystals, m.p. 45.2°. 

Torula utilis 

R. L. Lester, F. L. Crane and Y. Hatefi, /. Am. Chem. Soc. 
80 4751(1958). (Isolation) 

F. W. Heaton, J. S. Lowe and R. A. Morton, J. Chem. Soc, 
4094 (1956). 

b. NAPHTHOQUINONES 

516 Flaviolin, CioH^Og, garnet red rhombs containing solvent of 

crystallization, m.p.: dec. near 250°. 




Aspergillus citricus (Wehmer) Mosseray 
J. E. Davles, F. E. King and John C. Roberts, Chem,. and 
Ind., 1110 (1954). (Structure) 

517 6-Methyl-l,4-naphthoquinone, CnHgOa, golden yellow needles, 
m.p. 90-91°. 

O 



CH3 II 

Marasmius gramineum Lib. 



249 Naphthoquinones 

Gerd Bendz, Acta Chem. Scand. 2 192 (1948). 
Idem., ibid. 5 489 (1951). 

518 Phthiocol, CiiHsOg, yellow prisms, m.p. 173-174°. 

? CHs 



11 OH 

Mycobacterium tuberculosis var. hominis, Corynebac- 
terium diphtheriae 

R. J. Anderson and M. S. Newman, J. Biol. Chem. 103 197 
(1933). 

Rudolph J. Anderson, R. L. Peck and M. M. Crelghton, ibid. 
136 211 (1940). 

Michizo Asano and Hideo Takahashi, /. Pharm. Soc. Japan 
65 17 (1945). 

M. Terni, Boll. soc. ital. biol. sper. 25 60 (1949). 

There is evidence that phthiocol is an artifact, and that 
the precursor is a compound related to vitamin K,, but of 
higher molecular v^^eight. 

J. Francis, J. Madinaveitia, H. M. Macturk and G. A. Snow, 
Nature 163 365 (1949). 

519 Mollisin, C14H10O4CI2, orange-yellow needles, m.p. 202° (dec). 

HO O 

I II CI 




^"^ CH. & ^' 

I 

c=o 

CH3 

Mollisia caesia, Sacc. sensu Sydow, M. gallens Karst. 
G. J. M. van der Kerk and J. C. Overum, Rec. trav. chim. 
76 425 (1957). 

520 Javanicin, C15H14O6, red laths, m.p. 208°. 

OH O 
CH3O I II CH2COCH3 




Pfizer Handbook of Microbial Metabolites 250 

Fusarium javanicum Koorders 

Yield about 20 mg. per liter (purified pigment). Oc- 
curs together with fusarubin. 

H. R. V. Arnstein and A. H. Cook, J. Chem. Soc, 1021 
(1947). (Isolation) 

521 Fusarubin (Oxyjavanicin), Ci.-,Hi407, red prisms, m.p. 218° 
(preheated block). 



CH3O 




Fusarium solani (Mart.) App. and Wr. 

Yield about 50 mg. per liter (mixed with javanicin). 

H. R. V. Arnstein and A. H. Cook, /. Chem. Soc, 1021 
(1947). 

Hans W. Ruelius and Adeline Gauhe, Ann. 569 38 (1950). 

After ether extraction of the acidified broth, a water- 
soluble derivative of fusarubin remains behind. This has 
been identified as a sulfate ester occurring at one of the 
hydroquinone hydroxyl groups and was called fusarubino- 
gen. Fusarubinogen actually is present in the broth in a 
reduced form, which is probably a derivative of ^-hydro- 
naphthazarin. 

Hans W. Ruelius and Adeline Gauhe, Ann. 570 121 (1951). 

522 Bostrycoidin, C1SH14O7 (proposed), red or brown lath clusters, 

m.p. 243°. 

A substituted naphthoquinone similar to javanicin. 

Fusarium bostrycoides Wr. and Rkg. 

Mary Alice Hamilton, Marjorie S. Knorr and Florian A. 
Cajori, Antibiotics and Chemotherapy 3 853 (1953). 

F. A. Cajori, Theodore T. Otani and Mary Alice Hamilton, 
J. Biol. Chem. 208 107 (1954). (Isolation) 

523 4,9-Dihydroxyperylene-3,10-quinone, C00H10O4, dark red needles, 

dec. near 350°. 



o=< >=( >=o 



Daldinia concentrica (Bolt) Ces. and de Not. 



251 Naphthoquinones 

J. M. Anderson and J. Murray, Chem. and Ind., 376 (1956). 
(Isolation) 

It has since been reported that this perylenequinone is 
524 probably an artifact of 4,5,4'5'-tetrahydroxy-l,r-dinaphthyl. 



This polyphenol was obtained from the same organism. 
It was found to oxidize in part to a dark, melanin-like 
polymer, and in part to the perylenequinone. The struc- 
ture was proved by synthesis. 

J. D. Bu'Lock and D. C. AUport, Proc. Chem. Soc, 264 
(1957). 

D. C. Allport and J. D. Bu'Lock, /. Chem. Soc, 654 (1960). 

525 Mycochrysone, C20H14O7, orange-red crystals, m.p. : slow dec. 
above 180°. 

No N, — OCH3, C— CH3 nor halogen. Three active hy- 
drogens. 

Partial structure: 




-OH (phenolic or enolic) 
-H12O3 



An inoperculate discomycetous fungus. 
G. Read, P. Shu, L. C. Vining and R. H. Haskins, Can. J. 
Chem. 37 731 (1959). 

526 Actinorhodin, C32H26-30O14, bright red needles, dec. 270°. 



R,] 2COOH 

R2 i,2CH3 

R3 I C8Hi2_16 

' HO O O OH 

Streptomyces coelicolor (Miiller) Waksman and Hen- 
rici 




Pfizer Handbook of Microbial Metabolites 



252 



The yield was about 15% of the mycelial weight. 

Hans Brockmann and Ernst Hieronymus, Chem. Ber. 88 
1379 (1955). 

This compound has been shown to be an artifact, and 
by careful isolation under acidic conditions the precursor, 
protoactinorhodin, with the nucleus below, can be isolated. 




OH OH 



OH OH 




OH OH 



-> Actino- 
rhodin 



527 Protoactinorhodin was isolated as pale red prisms, dec. near 

330°, probably C30H30O14. 

Hans Brockmann and Volkmar Loeschcke, Chem. Ber. 88 
778 (1955). 

528 Xylindein, C34H26O11, deep brown high-melting, pleochroic leaf- 

lets. 

The structure is obscure, but an extended quinone sys- 
tem of the type 



was postulated. 

Chlorosplenium aeruginosum (Oeder ex Fries) De Not 
Fritz Kogl and G. von Taeuffenbach, Ann. 445 170 (1925). 
Fritz Kogl and Hanni Erxleben, ibid. 484 65 (1930). 

529 Rhodomycetin, gradual darkening at 300°. 

Dark fed powder, red in acid solution and blue in alka- 
line. U.V. 235, 540, 580 m^x. 

Reddish violet in H0SO4, positive FeCla, H0O2 and 
Na2So04-2HoO reduction. 

Resembles actinorhodin. 

Streptomyccs griseus 

Gerald Shockman and Selman A. Waksman, Antibiotics 
and Chemotherapy 1 68 (1951). 

530 Naphthoquinone from Mycobacterium phlei, yellow oil, U.V. 243, 

249, 261, 270, 328 m^ in isooctane. 

Appears to have about 30 carbon atoms and is probably 
a vitamin K,. Mol. wt. about 620. 



^53 



Naphthoquinones 



Mycobacterium, phlei 

Ten mg. were obtained from 450 g. of wet cells. 
A. F. Brodie, B. R. Davis and L. G. Fieser, /. Am. Chem. 
Soc. 80 6454 (1958). 

Vitamins K^: 

Vitamin Ko was first isolated from putrefied fish meal In 
1939 by Doisy and collaborators. Tishler and Sampson 
later found that it was produced by pure cultures of Bacil- 
lus brevis. Isler and collaborators corrected the structure 
originally proposed to A. below. They also isolated a 
lower isoprenolog, B., from putrefied fish meal. Both 
structures were proved by synthesis. The later group also 
determined the structure of (and synthesized) a higher 
isoprenolog C. isolated earlier in England. 



531 A., CiQH(i^0.2, hght yellow plates, m.p. 54°. 



CHs 

^CH2— CH=C— 

I 
CH3 



— CH2— CH2— CH=C — 

I 
CH3 



-CHs 



Bacillus brevis 

R. W. McKee, S. B. Binkley, Sidney A. Thayer, D. W. Mac- 
corquodale and Edward A. Doisy, J. Biol. Chem. 131 327 
(1939). 

M. Tishler and W. Sampson, Proc. Soc. Exp. Biol. 68 136 
(1948). 

532 B., C41H56O2, hght yellow plates, m.p. 50°. 
O 




CH2— CH=C- 



CH3 



-CH2— CH,— CH=C— 

I 
CH3 



— CH3 



O. Isler, R. Riiegg, L. Chopard-dit-Jean, A. Winterstein and 
O. Wiss, Helv. Chim. Acta 41 786 (1958). 



Pfizer Handbook of Microbial Metabolites 254 

533 C. CseHgoOo, yellow crystals, m.p. 58-59°. 
O 




CHo— CH=C— — CH2— CH2— CH=C— 1— CH3 

CH3 CH3 8 

Mycobacterium tuberculosis (Brevannes) 

This substance constituted about 0.59c of the dry cell 
weight. 

J. Francis, J. Madinaveitia, H. M. Macturk and G. A. Snow, 
Nature 163 365 (1949). (Isolation) 

H. Noll, R. Riiegg, U. Gloor, G. Ryser and O. Isler, Helv. 
Chim. Acta 43 433 (1960). (Structure and synthesis) 

C. ANTHRAQUINONES 

534 Anthraquinone pigment from Gibberella fujikuroi, probably 
C14H10O7, red crystals, m.p. 325° (sealed tube). 
Partial and tentative structure: 

CHoOH 

+ 20H 




O OH 

The structure may resemble that of cynodontin. 
Gibberella fujikuroi (Saw.) Wollenweber 
Yukihiko Nakamura, Tokuji Shimomura and Joji Ono, 
J. Agr. Chem. Soc. Japan 31 669 (1957). (Isolation) 

535 Clavorubin, C14H12O9, red crystals. 

Has one C — CH3 group. U.V. absorption resembles a 
1,5,8-trihydroxyanthraquinone. The leuco-acetate (like 
that of chrysergonic acid) has a diphenyl-like absorption. 

Claviceps purpurea 

B. Franck and T. Reschke, Angew. Chem. 71 407 (1959). 

536 Emodic Acid, C].-,Hs07, orange needles, m.p. 363-365°. 

O 

COOH 




^'JO 



Anthraquinones 



Penicillium cyclopium Westling 

Winston K. Anslow, John Breen and Harold Raistrick, 
Biochem. J. 34 159 (1940). 

537 Boletol, Ci-.HsO-, red needles, m.p. 275-280° (dec). 
O COOH ._ HO O 





HO O HO O COOH 

Boletus luridus Schaeff. ex Fries, B. badius Ft., B. 
chrysenteron Bull., B. satanas Lenz, B. subtomentosus 
Linn. 

The higher yielding species gave about 1 g. of pure 
material from 20 kg. of fruiting body. 

Fritz Kogl and W. B. Deijs, Ann. 515 10, 23 (1935). (Syn- 
thesis ) 

538 Pachybasin, C15H10O3, yellow needles, m.p. 78°. 

O 




Pachybasium candidum (Sacc.) Peyronel 
Pachybasin, like most of the other anthraquinone pig- 
ments, occurs as one constituent of a mixture of pigments. 
Chrysophanol was identified as one of the other con- 
stituents of this mixture. 

Shoji Shibata and Michio Takido, Pharm. Bull. (Tokyo) 3 
156 (1955). 

539 Chrysophanol ( Chry sophanic Acid), C15H10O4, dark yellow leaf- 
lets, m.p. 196°. 

O 




Penicillium islandicum Sopp, Pachybasium candidum 
(Sacc.) Peyronel, Chaetomium affine Corda 

The 9-anthrone corresponding to chrysophanol has been 
isolated from higher plants. 



Pfizer Handbook of Microbial Metabolites 256 

B. H. Howard and H. Raistrick, Biochem. J. 46 49 (1950). 
Shoji Shibata, Kagaku (Science) 26 391 (1956). 

540 Islandicin, C15H10O5, dark red plates, m.p. 218°. 

O OH 




Penicillium islandicum Sopp. 

This mold produces a complex mixture of pigments 
constituting up to 20% of the mycelial weight. 

B. H. Howard and H. Raistrick, Biochem. J. 44 227 (1949). 

Islandicin seems to be identical with funiculosin, a 
trihydroxyanthraquinone pigment of the same melting 
point and empirical formula isolated from Penicillium 
funiculosum Thom, a species closely related to P. is- 
landicum. 

Hisanao Igarasi, J. Agr. Chem. Soc. Japan 15 225 (1939). 

541 Helminthosporin, CigHioOg, dark maroon needles, m.p. 227°. 

"? ? CH. 



HO O OH 

Helm,inthosporium gramineum Rabenhorst, H. cyno- 
dontis Marignoni, H. catenarium, H. triticivulgaris 
Nisikado 

About 30% of the dry myceUum of H. gramineum con- 
sisted of anthraquinone pigments, mainly helmintho- 
sporin and catenarin. 

Harold Raistrick, Robert Robinson and Alexander R. Todd, 
J. Chem. Soc, 488 (1933). 

542 Emodin (Frangula-Emodin), C15H10O5, orange needles, m.p. 
255°. 




257 Anthraquinones 

Cortinariits sanguineus (Wulf. ) Fries, Chaetomium 
affine Corda. 

A yield of about S'c of the dry mycelial weight has been 
mentioned. 

Fritz Kogl and J. J. Postowcky, Ann. 444 1 (1925). 

R. A. Jacobson and Roger Adams, J. Am. Chem. Soc. 46 
1312 (1934). (Synthesis) 

543 Versicolorin, Ci^HjoOe, yellow-orange needles, m.p. 282°. 

HO O OH ^^^^ HO O OH ^^ ^^ 

I II I CH2OH I II I CH2OH 



II OH HO II 

O O 

Aspergillus versicolor (Vuillemin) Tiraboschi 

The same organism produces an uncharacterized xan- 

thone pigment. 

Yuishi Hatsuda and Shlmpei Kuyama, J. Agr. Chem. Soc. 

Japan 29 11 (1955). 

544 Cynodontin, Cj^HioOe, bronze plates, m.p. 260°. 

HO O OH 




HO O OH 

Helminthosporium cynodontis Marignoni, H. euclaenae 
Zimmermann, H. avenae Ito and Kurib, H. victoriae 

Winston Kennay Anslow and Harold Raistrick, Biochem. J. 
34 1546 (1940). (Synthesis) 

545 oj-Hydroxyemodin ( Citreorosein ) , CjgHioOe, orange needles, 
m.p. 288°. 

n 

CH2OH 




Penicillium cyclopium Westling, P. citreo-roseum 
Dierckx. 

Winston K. Anslow, John Breen and Harold Raistrick, 
Biochem. J. 34 159 (1940). 



Pfizer Handbook of Microbial Metabolites 258 

Theodore Posternak, Compt. rend. soc. phys. his. nat. Ge- 
neve 56 28 (1939). 

546 Catenarin, Ci^HiyOe, red plates, m.p. 246°. 

HO ° °" 




Helminthosporium catenarium Drechsler, H. grami- 
neum Rabenhorst, H. velutinum Link, H. triticivulgaris 
Nisikado, Penicillium islandicum Sopp, Aspergillus am- 
stelodami (Mangin) Thorn and Church 

More than 15% of the mycelial weight of H. catenarium 
was catenarin. 

Winston Kennay Anslow and Harold Raistrick, Biochem. J. 
35 1006 (1941). (Synthesis) 

547 Asperthecin, Ci^HioO^j, chestnut brown needles, no m.p. 

O HO O 

CH.OH HO 1 I! CH2OH 





Aspergillus quadrilineatus Thom and Raper and other 
species of the Aspergillus nidulans group 

S. Neelakantan, Anna Pocker and H. Raistrick, Biochem. J. 
66 234 C1957). 

A closely related pigment has been observed, which may 
have been a tautomeric or reduced form of asperthecin. 
It could not be isolated because of its ready conversion to 
asperthecin. 

B. H. Howard and H. Raistrick, Biochem. J. 59 475 (1955). 

548 Fallacinal, Ci«Hi„0,., orange-yellow needles, m.p. 251°. 
CH3O ? CHO 




259 Anthraquinones 

Xanthoria fallax (Hepp.) Arn. 

Takao Murakami, Pharm. Bull. (Tokyo) 4 298 (1956). 

549 Tritisporin ( to-Hydroxycatenarin ) , Cjr.HioO;, brown needles, 
m.p. 260-262°. 

°" CH,OH 




Helminthosporium triticivulgaris Nisikado 
S. Neelakantan, Anna Pecker and H. Raistrick, Biochem. J. 
64 464 (1956). 

550 Flavoskyrin, Ci.-,Hi^O.-,, yellow crystals, m.p. 208° (dec), [cf]u 
-295° (in dioxane). 




PenicilHvm islandiciim Sopp. 

Shoji Shibata, Takao Murakami and Michio Takito, Pharm. 
Bull. (Tokyo) 4 303 (1956). (Structure) 

551 Compound A ( 1,4,7, 8-Tetrahydroxy-2-methylanthraquinone), 
Ci,Hi,0,. 

An optically inactive compound (no melting point 
given). Treatment with cone. H2SO4 yields an anthraqui- 
none, Ci.-Hi„0.,, red crystals, m.p. 255°, with the follow- 
ing structure: 




O 

Penicillium islandicum 

Sten Gatenbeck, Acta Chem. Scand. 12 1985 (1958). 

Idem., ibid. 13 705 (1959). 



Pfizer Handbook of Microbial Metabolites 260 

552 Endocrocin, CieHjoOj, copper-red leaflets, m.p. 318° (dec.)- 

O 

HO II CH3 



I II I COOH 
HO O OH 

Nephromopsis endocrocea Asahina 

Yasuhiko Asahina and Fukuziro Fuzikawa, Ber. 68B 1558 
(1935). 

Aspergillus amstelodami (Man gin) Thorn and Church. 

Shoji Shibata and Shinsaku Natori, Pharm. Bull. (Tokyo) 1 
160 (1953). 

553 Clavoxanthin, CieHioOy, yellow needles, m.p. 340° (dec). 

Apparently similar to endocrocin. 

Claviceps purpurea 

B. Franck and T. Reschke, Angew. Chem. 71 407 (1959). 

554 Parietinic Acid, Ci^HioOy, yellow needles, m.p. --'300° (sub- 

limes). 



CH3O II COOH 




Xanthoria parietina (L.) Th. Fr. 

Walter Escherich, Biochem. Z. 330 73 (1958). 

555 Physcion (Partetin), CigHisOg, orange-yellow leaflets, m.p. 207°. 

O 
CH3O 




Aspergillus glaiicus spp., A. chevalieri, A. ruber 
(Mangin) Raper and Thom, Penicillium herquei Bainier 
and Sartory, Xanthoria parietina (L.) Beltram, X. fallax, 
Teloschistes fiavicans (Sw.) Norm., T. exilis Wainio, 
Placodium spp., Caloplaca elegans (Link) 



26 1 Anthraquinones 

F. Rochleder and W. Heldt, Ann. 48 1 (1843). 

Harold Raistrick, Enzymologia 4 76 (1937). 

H. Raistrick, Robert Robinson and A. R. Todd, /. Chem. 
Soc, 80 (1937). 

Julius Nicholson Ashley, Harold Raistrick and Taliesin 
Richards, Biochem. }. 33 1291 (1939). 

T. R. Seshadri and S. Sankara Subramanian, Proc. Indian 
Acad. Sci. 30A 67 (1949). 

Walter B. Mors, Bol. Inst. Quim. Agric. No. 23 7 (1951). 

S. Neelakantan and T. R. Seshadri, /. Sci. Ind. Research 
(India) IIB 126 (1952). 

Shoji Shibata and Shinsaku Natori, Pharm. Bull. (Tokyo) 1 
160 (1953). 

Mitizo Asano and Yosio Arata, /. Pharm. Soc. Japan 60 521 
(1940). 

J. A. Galarraga, K. G. Mill and H. Raistrick, Biochem. J. 61 
456 (1955). 

Jiro Kitamura, Uzuhiko Kurimoto and Matatsugu Zoko- 
yama, /. Pharm. Soc. Japan 76 972 (1956). 

556 Macrosporin, CjfiHisO^, orange-yellow rhombic crystals, m.p, 
300° (dec). 



CH3O 




Macrosporium porri Elliott 

R. Suemitsu, Y. Matsui and M. Hiura, Bull. Agr. Chem. Soc. 
(Japan) 21 1-4, 337 (1957). (Isolation) 

R. Suemitsu, M. Nakajima and M. Hiura, ibid. 23 547 
(1959). 

557 Teloschistin (Fallacinol), CigHisOe, orange plates, m.p. 245- 
247°. 



CH3O II CH2OH 




Teloschistes flavicans (Sw. ) Norm., Xanthoria fallax 
(Hepp.) Am. 



Pfizer Handbook of Microbial Metabolites 262 

T. R. Seshadri and S. Sankara Subramanian, Proc. Indian 
Acad. Sci. 30A 67 (1949). 

558 Roseopurpurin (Carviolin), CieHjoOg, yellow needles, m.p. 
286°. 



CH2OH 




HO O OCH3 
Penicillium roseopurpureum Dierckx 

559 A second pigment, carviolacin, CooHjeO;, light brown 
needles, m.p. 243° (dec), was isolated from this mold. 
It is apparently closely related in structure. 

Theodore Posternak, Helv. Chim. Acta 23 1046 (1940). 
H. G. Hind, Biochem. J. 34 67, 577 (1940). 

560 Erythroglaucin (Catenarin 6-Methyl Ether), CigHisOg, deep red 

plates or needles, m.p. 205°. 

CH3O 




Aspergillus glaucus (ten spp.) 

The former rubroglaucin was shown to be a mixture of 
physcion and erythroglaucin. 

Julius Nicholson Ashley, Harold Raistrick and Taliesin 
Richards, Biochem. J. 33 1291 (1939). 

561 Neophromin, CieHisO,,, ocher colored needles, m.p. 198° (dec). 

A quinone-like pigment. 

Neophromium lusitanicinn 

O. Hesse, J. prakt. Chem. 57 409 (1898). 

562 Dermocybin, C^^-H^.^O-, red needles, m.p. 228°. 

This is an incompletely characterized anthraquinone 
pigment. It has five nuclear hydroxyl groups, one of 
them methylated. It is produced along with emodin by 



^^3 Anthraquinones 

Cortinarius sanguineus (Wulf.) Fries and constitutes 
0.2-0.4% of the mycelial weight. 

Cortinarius cinnabarinus Fries produces a pigment 
which is the same or similar. 

Fritz Kogl and J. J. Postowsky, Ann. 444 1 (1925). 

563, 564 Physcion Anthranols, Ci,jHi404, m.p.'s 260° and 181°. 

CHsO^ II ,CH3 CH3O CH3 

and 




OH HO O OH 

Aspergillus glaucus (five types) 

Julius Nicholson Ashley, Harold Raistrick and Tallesin 
Richards, Biochem. J. 33 1291 (1939). 

565 Rhodocladonic Acid, Ci^Hj.Oc,, red needles, m.p. >360°. 

O 

'^ 

\ 

COOCHj 

Thirteen Cladonia species 

Shoji Shibata, Michio Takido and Osamu Tanaka, /. Am 
Chem. Soc. 72 2789 (1950). 

566 Nalgiolaxin, CisHi-OgCl, yellow plates or needles, m.p. 248° 

[a].579o" +40.3° (in chloroform). 




H3O 




Penicillium nalgiovensis Laxa 

H. Raistrick and J. Ziffer, Biochem. J. 49 563 (1951). 



Pfizer Handbook of Microbial Metabolites 264 

567 Nalgiovensin, CigHisOg, orange needles or plates, m.p. 199- 
200°, [a]579o'° +39.7° (in chloroform). 

OH 
CH3O II CH2— CH— CH3 




Penicillium nalgiovensis Laxa 

H. Raistrick and J. ZifFer, Biochem. J. 49 563 (1951). 
(Isolation) 

A. J. Birch and R. A. Massy-Westropp, /. Chem. Soc, 2215 
(1957). (Structure) 

568 Thermophillin, CisHigOg, golden plates, m.p. subl. 245° (dec. 

260° sealed tube). 
Quinonoid properties. 
Lenzites thermophila 
H. S. Burton, Nature 166 570 (1950). 

569 Phomazarin,* CigHiYOgN, orange needles, m.p., 197° (dec). 

CH3CH2CH2CH2 O CH3CH2CH2CH" O 

CHaO^ I II /COOH CH3O 




I II OH I II I COOH 

HO O HO O OH 

Phoma terrestris Hansen 

F. Kqgl and J. Sparenburg, Rec. trav. chim. 59 1180 
(1940).' 

F. Kogl and F. S. Quackenbush, ibid. 63 251 (1944). 

F. Kogl, G. C. van Wessem and O. I. Elsbach, ibid. 64 23 
(1945). (Synthesis) 

570 Atrovenetin, CigHigOe, brownish yellow prisms, m.p. 295° 
(dec), [a]546i'' +154° (c 0.486 in dioxan). 




Penicillium atrovenetum G. Smith 
* See addendum. 



265 Anthraquinones 

K. G. Neill and H. Raistrick, Chem. and Ind., 551 

(1956). (Isolation) 

Idem., Biochem. J. 65 166 (1957). (Isolation) 
D. H. R. Barton, P. de Mayo, G. A. Morrison and H. Rai- 
strick, Tetrahedron 6 48 (1959). (Structure) 

571 Norherqiieinone, C19H1SO7, dark red needles, m.p. 279° (dec), 

[a],r' +1080° ±60° (c 0.048 in pyridine). 
Structure: Unmethylated herqueinone 
See herqueinone for organism, structure and references. 

572 Herquein, CigHooOg (proposed), yellow-brown crystals, m.p. 

129° (decO. 
Water-soluble. Fluoresces in alkali. 
Penicillium herqiiei 
H. Stowar Burton, Brit. J. Exptl. Path. 30 151 (1949). 

573 Herqueinone, CooHoqOj, red needles, m.p. 226° (dec.) (sub- 

limes), [aW- +440° ±40° (c 0.063 in ethanol). 
Partial structure: 




Penicillium herquei Bainier and Sartory 

A crude pigment yield of 17% of the weight of the dry 
mycelium was obtained. The major constituents were 
norherqueinone and its methyl ether, herqueinone. Mi- 
nor constituents were physcion and meso-erythritol. 

The plant pigment, haemocorin, also contains the peri- 
naphthenone nucleus. 

Frank H. Stodola, Kenneth B. Raper and Dorothy 1. Fen- 
neU, Nature 167 773 (1951). (Isolation) 

J. A. Galarraga, K. G. NeUl and H. Raistrick, Biochem. J. 61 
456 (1955). 

D. H. R. Barton, P. de Mayo, G. A. Morrison, W. H. Schaeppi 
and H. Raistrick, Chem. and Ind., 552 (1956). (Structure) 

Robert E. Harman, James Cason, Frank H. Stodola and 
A. Lester Adkins, /. Org. Chem. 20 1260 (1955). 

574 Solorinic Acid, C01H20O7, red-brown plates, m.p. 203.5°. 
CH3O 11 OH 



\ 

CO(CH2)4CH3 




OH 



Pfizer Handbook of Microbial Metabolites 266 

Solorina crocea (L. ) Ach. 

G. Roller and H. Russ, Monatsh. 70 54 (1937). 

575 Resistomycin, CouHi^Og, yellow needles, m.p. 315° (dec.) (sub- 
limes from 215°). 



CH3— 




OH O 



Streptomyces resistomycificus 

Hans Brockmann and Giinter Schmidt-Kastner, Chevi. Ber. 
87 1460 (1954). (Isolation) 

H. Brockmann, E. Meyer and K. Schrempp, Dissertations, 
University of Gottingen, 1954, 1958. (Partial structure by 
courtesy of Prof. Brockmann) 

576 Granatacin, C20H20O10, pomegranate-red crystals, m.p. 204- 

206° (dec). 

A tricyclic tetrahydroxyquinonedicarboxylic acid with 
antibiotic properties. 

Streptomyces olivaceus (Waksman) Waksman and 
Henrici 

R. Corbaz, L. Ettlinger, E. Gaumann, J. Kalvoda, W. Keller- 
Schierlein, F. Kradolfer, B. K. Manukian, L. Neipp, V. Prelog, 
P. Reusser and H. Zahner, Helv. Chim. Acta 40 1262 (1957). 

577 Luteomycin (Antibiotic 289), C2cH;5;.OioN (proposed), (Hydro- 

chloride) orange-yellow crystals, m.p. 199° (dec). 

Color changes to purple in alkali. Positive quinone- 
Na^COg, FeClg. Negative ninhydrin, biuret, MoUsch, 
Fehling, Sakaguchi. Can be precipitated as reineckate, 
helianthate or pier ate. 

Streptomyces flaveolus, S. tanashiensis related to S. an- 
tibiotic us 

Toju Hata, Tomojiro Higuchi, Yoshimoto Sano and Katuko 
Sawashi, Kitasato Arch. Exptl. Med. 22 229 (1949). 

Hamao Umezawa, Tomio Takeuchi, Kazuo Nitta, Kenji 



267 



Anthraquinones 



Maeda, Tadashi Yamamoto and Seizaburo Yamaoka, /. Anti- 
biotics (Japan) 6A 45 (1953). 

Teisuke Osato, Koki Yagishita, Ryozo Utahara, Masahiro 
Ueda, Kenji Maeda and Hamao Umezawa, ibid. 6A 52 (1953). 

Berislav Govorcin, Tehnicki Pregled 8 43 (1956). 

578 Luteoleersin, C.uH.isOy, yellow crystals, m.p. 135°, [(z]r,46i" 214° 

(c 0.456 in ethanol). 

Believed to be a substituted quinone, containing two 
active hydrogens. It was accompanied by a reduction 
product : 

579 Alboleersin, CoeH^nO^, colorless crystals, m.p. 215°, [a].r546i^^ 

274° (c 6.430 in ethanol). 

Contains three active hydrogens. 

Helmiiithosporiuin leersii Atkinson 

Julius N. Ashley and Harold Raistrick, Biochem. J. 32 449 
(1938). 

580 Skyrin (Endothianin), C;^„HisOt,i, dark orange rods, m.p. 

>380°. 




Penicillium islandicum Sopp, P. wortmanni Klocker, P. 
tardum Thom, P. rugulosum Thorn, Endothia parasitica 
(Murr. ) Anderson and Anderson, E. fiuens Shear and 
Stevens 

All of these fungi produce a mixture of skyrin with 
rugulosin. 

F. Kogl and F. S. Quackenbush, Rec. trav. chim. 63 251 
(1944). 



Pfizer Handbook of Microbial Metabolites 



268 



Shoji Shibata, Osamu Tanaka, Goro Chihara and Horoshi 
Mitsuhashi, Pharm. Bull. (Tokyo) 1 302 (1953). 

Shoji Shibata, Takao Murakami, Osamu Tanaka, Goro Chi- 
hara, Isao Kitagawa, Masashi Sumimoto and Chikara Kaneko, 
ibid. 3 160 (1955). (Structure) 

Shoji Shibata, Takao Murakami, Osamu Tanaka, Goro Chi- 
hara and Masashi Sumimoto, ibid. 3 274 (1955). 

J. Breen, J. C. Dacre, H. Raistrick and G. Smith, Biochem. J. 
60 618 (1955). 

Shoji Shibata, Michio Takido and Terumi Nakajima, 
Pharm. Bull. (Tokyo) 3 286 (1955). 

Yuzuru Yamamoto, Takeo Yamamoto, Skoichi Kanatomo 
and Kiyoshi Tanimichi, /. Pharm. Soc. Japan 76 192 (1956). 

Yazuru Yamamoto, Akira Hamaguchi, Isao Yamamoto and 
Sumie Imai, ibid. 76 1428 (1956). 



581 Pigment B: CsoHigOn. 




CH2OH 



HO O OH 



582 Pigment C: CgoHigOia. 



HO O OH 




CHoOH 



CH2OH 



HO O OH 
These are oxidized skyrins. 



269 



Anthraquinones 



Penicilliiim islandicuvi N.R.R.L. 1175 
Shoji Shibata, Michio Takido and Terumi Nakajima, 
Pharm. Bull. (Tokyo) 3 286 (1955). 

583 Iridoskyrin, CgoHigOig, irridescent red rods or plates, m.p. 358°. 

HO O OH 




HO O OH 

Penicillium islandicum Sopp. 

B. H. Howard and H. Raistrick, Biochem. J. 57 212 (1954). 

584 Aurofusarin, CyijHooOio, m.p. >360°. 

This Incompletely characterized pigment produced by 
Fusarium culinorum W. G. Smith may be a dianthraqui- 
none. 

Julius N. Ashley, Betty C. Hobbs and Harold Raistrick, 
Biochem. J. 31 385 (1937). 

585 Penicilliopsin, C30H22O8, orange crystals, m.p. 330° (dec.)- 

HO O OH 




Penicilliopsis clavariaeformis Solms-Laubach 

H. Brockmann and H. Eggers, Angew. Chem. 67 706 (1955). 



Pfizer Handbook of Microbial Metabolites 



270 



586 Rugulosin (Radicalisin), C30H00O10, yellow prisms, m.p. 293' 
(dec), [a]546i'' +605° (dioxane). 




Penicillium rugulosum Thorn, P. tardum Thorn, P. 
wortmanni Klocker, Endothia parasitica (Murr. ) Ander- 
son and Anderson, E. fiiiens Shear and Stevens. 

About 20% of the dry weight of P. rugulosum myce- 
lium is rugulosin. 

J. Breen, J. C. Dacre, H. Raistrick and G. S. Smith, 
Biochem. J. 60 618 (1955). 

Shoji Shibata, Osamu Tanaka, Goro Chihara and Horoshl 
Mitsuhashi, Pharm. Bull. (Tokyo) 1 302 (1953). 

Shoji Shibata, Takao Murakami, Osamu Tanaka, Goro Chi- 
hara, Isao Kitagawa, Masashi Sumimoto and Chikara Kaneko, 
ibid. 3 160 (1955). (Structure) 

Shoji Shibata, Takao Murakami, Osamu Tanaka, Goro Chi- 
hara and Masashi Sumimoto, ibid. 3 274 (1955). 

Yazuru Yamamoto, Akira Hamaguchi, Isao Yamamoto and 
Sumie Imai, J. Pharm. Soc. Japan 76 1428 (1956). 

Shoji-Shibata and Isao Kitagawa, Pharm. Bull. (Tokyo) 4 
309 (1956). (Structure) 

587 Rubroskyrin, C30H22O12, dark red plates, m.p. 289° (dec). 

O O OH 




271 Anthraquinones 

Penicilliinn islaudiciim Sopp. 

This pigment is produced in a mixture including is- 
landicin, iridoskyrin, erythroskyrin, catenarin, luteoskyrin 
and skyrin. The weight of the pigment mixture is about 
10% of the weight of the dry mycelium. 

Shoji Shibata and Isao Kitagawa, Pharm. Bull. (Tokyo) 4 
309 (1956). 

588 Luteoskyrin, C;^oH^,._.0,^,, yellow needles, m.p. 273° (dec), [aln^^ 
-880° (in acetone). 



HO 



OH 



HO 
HO 



O OH 
O OH 



CHs 



CH3 



^1 



HO 



OH 




Penicillium islandicum Sopp. 

Shoji Shibata and Isao Kitagawa, Pharm. Bull. (Tokyo) 4 
309 (1956). (Structure) 

589 Cercosporin, C^oH.sO^o, red crystals, m.p. 241°, [a]:,,,,!.'" +470° 
(c 0.5 in chloroform). 

This pigment contains two methoxyl groups, two 
quinoid carbonyls, two phenolic hydroxyls and two alco- 
holic hydroxyls. The yield was 79 mg. per gram of dry 
mycelium. 

Shimpei Kuyama and Teiichi Tamura, /. Am. Chem. Soc. 
79 5725, 5726 (1957). 

3,591 Chaetochrysin and Chaetoflavin, C^iHoj-Oi,, yellow crystals, no 
592 melting point, and Chaetoalbin, CaoHo^.-joOj], white crys- 

tals, no melting point. 

These uncharacterized compounds were isolated from 
mycelial extracts along with chrysophanol. They seem to 
be modified dianthraquinones. They yield some chrysoph- 
anol on alkaline oxidation, contain one methoxyl group 
and have high optical rotations. 

Chaetomiiim affine Corda 

Vincent Arkley, F. M. Dean, Peter Jones, Alexander Robert- 
son and John Tetaz, Croat. Chem. Acta 29 141 (1957). 



Pfizer Handbook of Microbial Metabolites 272 

593 Rifomycin B, C39H51O14N, m.p. 160-164° (dec). 

A dibasic acid (pKs 2.8, 6.7). Probably a quinone 
(U.V. peaks at 400-460, also at 223, 234). 

Streptomyces mediterranean 

P. Sensi, A. Greco and R. Ballotta, 7th Annual Symposium 
on Antibiotics, Washington, 1959. 

594 Vinacetin, yellow platelets, m.p. 157°. 

Apparently quinoid. Positive FeClg, violet color in 
alkali, positive Molisch, Liebermann, Fehling. Negative 
ninhydrin, Millon, Sakaguchi. 

Streptomyces sp. 

Kyuzo Omachi, J. Antibiotics (Japan) 6A 73 (1953). 

595 Rhodophyscin, red leaflets, m.p. 260° (dec). 

A quinone-like substance. 

Physica endococcina 

Wilhelm Zopf, Ann. 340 276 (1905). 



13 



Tetracycline, Analogues and Related Sub- 
stances 



The tetracycline antibiotics display features indicative of an 
acetate origin. The oxygenation pattern is generally consistent 
as are the points of occurrence of methyl groups and halogen 
atoms. There is also at least a superficial resemblance to 
proved acetate derivatives such as the anthraquinones. So far 
the experimental evidence published concerning the biosyn- 
thetic origin of the tetracyclines has been limited, and some in- 
teresting obscurities remain. 

The general concept of an acetate-derived precursor in the 
sense of a polyketomethylene chain is, in the case of oxytetra- 
cycline as follows: 



./ 



C, lO] 

C C 



/ 



[O] 



\ 



Ci 



10]C 




/ 



c 

/ \ 



o o o o 

loi 

A 6-demethyltetracychne has been isolated from a fermentation 
broth, and tetracycline itself is a 5-deoxyoxytetracychne as well 
as a 7-dechlorochlortetracycUne; so sometimes some of the steps 
in the biosynthetic scheme are omitted. 



Pfizer Handbook of Microbial Metabolites 274 

The production by Streptomyces rimosus of oxytetracy- 
cline-X\ a modification of Terramycin in which there is an 
acetyl group instead of a carboxamide group at position 2, sup- 
ports the acetate theory since terramycin-X is more directly in 
the line of descent from a polyketomethylene chain (ten head 
to tail condensed acetate units) than is Terramycin itself. 

The dehydro derivatives which have been isolated- also may 
be considered as precursors of the other tetracyclines since the 
additional double bond may be the (as yet unreduced) result of 
an aldol type of condensative ring closure with elimination of a 
water molecule. 

More experimental work has been reported on the biosyn- 
thetic origin of oxytetracycline than on that of related sub- 
stances. Addition of C^Hj-methionine and 2-C^*-acetic acid to 
oxytetracycline-producing fermentations yields radioactive oxy- 
tetracycline (Terramycin). Quantitative degradation and 
counting studies show that methionine furnishes the C,5-methyl 
and the N-methyl groups. The radioactivity of the degradation 
fragments from the molecule which had incorporated 2-C'^- 
acetic acid indicated that most of the molecule is in quantitative 
agreement with the theoretical requirements for acetate deriva- 
tion.^' * 

Results are entirely consistent with formation of the ring 
skeleton at least from C- to C^o by head to tail linkage of acetate 
units. Glutamic acid has been considered as a possible precur- 
sor of part of the A-ring (carboxamide side-chain, carbon atoms 
2, 3, 4, 4a and the 4-amino nitrogen) and 2-C^ '-labeled glutamic 
acid yielded a labeled oxytetracycline.-^ Later evidence'' indi- 
cates that acetate also is capable of furnishing these carbon 
atoms although the level of activity in the A-ring seems to be 
somewhat lower than the theoretical, particularly in Terramycin 
isolated from older fermentations. The degradation fragments 

^ F. A. Hochstein, M. Schach von Wittenau, Fred W. Tanner, Jr. 
and K. Murai, /. Am. Chem. Soc. 82 (1960). (In press) 

- J. R. D. McCormick, Philip A. Miller, John A. Growich, Newell O. 
Sjolander and Albert P. Doerschuk, ibid. 80 5572 (1958). 

3 A. J. Birch, J. F. Snell and P. L. Thompson, ibid. 82 2402 (1960). 

4 A. J. Birch and P. L. Thompson, ibid. 82 (1960). (In press) 
^ J. F. Snell, R. L. Wagner, Jr. and F. A. Hochstein, Internal. 

Conf. on Peaceful Uses of Atomic Energy, 431 (1955); J. F. Snell, 
Symposium on Uses of Isotopes, Uniontown, Pa., 1957. 

'A. J. Birch and P. L. Thompson, ]. Am. Chem. Soc. 82 (1960). 
(In press) 



275 Tetracycline, Analogues and Related Substances 

from this portion of the molecule are not satisfactory for the 
clarification of the origin of the A-ring. It remains to be seen 
whether or not a less direct mechanism of acetate incorporation 
prevails in this area. 

The isolation and identification of oxytetracycline-X (2-acetyl- 
2-decarboxamidooxytetracycline), a lower potency antibiotic, 
from cultures of Streptoviyces rimosus, the Terramycin pro- 
ducer, seem to support in a general way the idea of the acetate 
derivation of ring A. It is tempting to speculate that oxytetra- 

CHj CH3 

CH3 OHOH^N 




cycline-X may be a precursor of oxy tetracycline, but this has not 
been proved. 

With the acetate theory as a guide, it is possible to extrapolate 
some predictions from the tetracyclines isolated and character- 
ized to date. It would seem probable that other mutations of 
the producing organisms might be obtained in which one minor 
biosynthetic step is blocked. Thus, retention of an oxygen atom 
at position 8 might be expected. Similarly, other tetracyclines 
lacking the Cfi-methyl and/or hydroxyl groups, the Ci2a-hydroxyl 
group and perhaps the N-methyl groups may be found. It is 
also possible that glycosides may be isolated as in the pyrromy- 
cinones. 

The pyrromycinones are produced by streptomyces species, 
and they bear some resemblance to the tetracyclines. The four 
linear rings appearing in various states of oxidation and the 
similarity in the number of carbon atoms make it seem that 
their biogenetic origin may be similar to that of the tetracy- 
clines. Apparently no experimental work has been published 
on this point. There is probably a close relationship among the 
pyrromycinones, rhodomycinones and quinocyclines. All of 
these pigments are found occasionally as glycosides, but no 
tetracycline glycosides have been reported yet. 

The rhodomycins are a complex of red pigments produced 
by Streptomyces purpurascens. The original complex was sepa- 



Pfizer Handbook of Microbial Metabolites 276 

rated into four components; rhodomycin A, isorhodomycin A, 
rhodomycin B and isorhodomycin B. The first three were iso- 
lated in the crystalline state. These substances contained nitro- 
gen, and, on mild acid hydrolysis, yielded an amino sugar, rho- 
dosamine, CgHi703N, plus the aglycones (rhodomycinones, 
isorhodomycinones ) . 

The same organism has yielded a number of other pigments 
which do not contain nitrogen. These also have been desig- 
nated rhodomycinones. Three of these, ^, e and iso-e have been 
obtained crystalline. It has been reported (no experimental 
details) that a y-rhodomycinone and six other rhodomycinones 
have been isolated "in substance" and that three others have 
been demonstrated by paper chromatography. The rhodomy- 
cinones seem to resemble the pyrromycinones, quinocyclines, 
cinerubins and rutilantinone. 

596 Rhodomycin A (Hydrochloride), C20H29O7NHCI, fine, dark red 

needles, m.p. 193° (dec.) (preheated block). 

Hans Brockmann and Use Borchers, Chem. Ber. 86 261 
(1953). 

597 Isorhodomycin A (Hydrochloride), CooHsgOgNHCl (proposed), 

m.p. 220°, [a]606-76o'' +268 ±30° (c 0.1 in methanol). 

Hans Brockmann and Peter Patt, Chem. Ber. 88 1455 
(1955). 

598 Rhodomycin B (Hydrochloride), C19H27O7NHCI, red prisms, 

m.p. 180°, [a]606-76o'' +174 ±10° (c 0.05 in methanol). 

An isorhodomycin B also was present. 

Hans Brockmann and Peter Patt, Chem. Ber. 88 1455 
(1955). 

599 ^-Rhodomycinone, C2oHi40g (proposed), dark red needles, m.p. 

225°. 

Hans Brockmann and Burchard Franck, Chem. Ber. 88 
1792 (1955). (Isolation) 

Hans Brockmann and P. Boldt, Naturwissenschaften 44 616 
(1957). (Revised empirical formula) 

600 €-Rhodomycinone, C2iH220g, thick red prisms, m.p. 185° (dec. 

at 208°) 

and 

601 £ -Isorhodomycinone, C20H20O9, dark red leaflets, m.p. 245° 

(dec). 



277 



Tetracycline, Analogues and Related Substances 



Hans Brockmann and Burchard Franck, Chem Ber 88 1792 
(1955). (Isolation) 
Other references: 
Hans Brockmann and Klaus Bauer, Naturivissenschaften 37 

Hans Brockmann, Klaus Bauer and Use Borchers Chem 
Ber. 84 700 (1951). 

Hans Brockmann and Enno Spohler, Naturivissenschaften 
42 154 (1955). (Characterization of rhodosamine) 

Hans Brockmann, German Patent 913,813 (1954). 

602 7-Chloro-6-demethyltetracycline, C,iH,iOsNoCl (isolated as the 
sesquihydrate), yellow crystals, m.p. 174-178° (dec ) 
[<x]^-' -258° (0.5% in 0.1 N sulfuric acid). ' ' 




CONH2 



Streptomyces aureofaciens Duggar (mutant) 

J. R. D. McCormick, NeweU O. Sjolander, Ursula Hirsh 

Ekner R. Jensen and Albert P. Doerschuk, /. Am. Chem Soc 

79 4561 (1957). 

603 6-Demethyltetracycline, CaiHssOgNoCl (isolated as the hydro- 
chloride hemihydrate), yellow crystals, m.p. 203-209° 
(dec), [aJD -259° (c 0.5 in 0.1 N sulfuric acid). 




CONH2 



Streptomyces aureofaciens Duggar (mutant) 

J. R. D. McCormick, Newell O. Sjolander, Ursula Hirsh, 

Elmer R. Jensen and Albert P. Doerschuk, /. Am. Chem. Soc 

79 4561 (1957). 



Pfizer Handbook of Microbial Metabolites 278 

604 7^-Pyrromycinone, C22H16O7, red needles, m.p. 236° (sublimes). 
OH O OH 




CH3 

XH3^ 

OH O COOCH3 

Streptomyces spp. 

Hans Brockmann and Werner Lenk, Chem. Ber. 92 1880 
(1959). (Structure) 

Hans Brockmann and Hans Brockmann, Jr., Naturwissen- 
sctiaften 47 135 (1960). (Revised structure) 

605 ^-Pyrromycinone, CooHooO^, orange-red needles, m.p. 216° (sub- 
limes), [air,.'" +74 ±6° (in chloroform). 

OH O 




CH 
OH O COOCH3 

Streptomyces spp. 

Brockmann and collaborators have isolated about a 
dozen pigments of this type from various unclassified 
streptomycetes. 

Hans Brockmann and Burchard Franck, Chem. Ber. 88 1792 
(1955). 

H. Brockmann, Luis Costa Pla and W. Lenk, Angew. Chem. 
69 477 (1957). 

H. Brockmann and P. Boldt, Naturwissenschaften 44 616 
(1957). 

Hans Brockmann and Werner Lenk, Chem. Ber. 92 1880 
(1959). (Structure) 

Idem., Naturwissenschaften 47 135 (1960). (Revised 
structure) 

606 c-Pyrromycinone (Rutilantinone), C2L.H2()Of,, orange-red needles, 
m.p. 213°, [aW +143 ±7° (c 1.0 in chloroform). 




XHo 
OH O COOCH3 



Streptomyces spp. 



279 



Tetracycline, Analogues and Related Substances 



607 



6-Pyrromycinone occurs as such and also as the chro- 
mophore of the antibiotics pyrromycin and the cinerubins. 
It is identical with rutilantinone. 

Hans Brockmann and Werner Lenk, Chem. Ber. 92 1880 
(1959). (Structure) 

Idem., Naturwissenschaften 47 135 (1960). (Revised 
structure) 

H. Brockmann, H. Brockmann, Jr., J. J. Gordon, W. Keller- 
Schierlein, W. Lenk, W. D. Ollis, V. Prelog and I. O. Suther- 
land, Tetrahedron Letters No. 8, p. 25 (1960). 

W. D. Ollis, I. O. Sutherland and J. J. Gordon, Tetrahedron 
Letters No. 16, p. 17 (1959). 

7-Chloro-5a(Ila)-dehydrotetracycline, C<,.H.iOsN^Cl, [ajo^' 15.5° 
(c 0.65 in 0.03 N hydrochloric acid). 




CONH2 
OH O O O 

Streptomyces aiireofaciens Duggar inutant 

The analogous compounds in which the chlorine atom 

is replaced by H and Br are also claimed. 

J. R. D. McCormick, Philip A. Miller, John A. Growich, 

Newell O. Sjolander and Albert P. Doerschuk, /. A?n. Chem. 

Soc. 80 5572 (1958). 

608 Chlortetracycline (Aureomycin, Biomycin), C22H23O8N2CI, fine 
yellow crystals, m.p. 168°, [aln'' -274.9° (in methanol). 

CH3 CH3 




CONH2 
OH O OH O 

Streptomyces aureofaciens 

R. W. Broschard, A. C. Dornbush, S. Gordon, B. L. Hutch- 
ings, A. R. Kohler, G. Krupka, S. Kuchner, D. V. Lefemine and 
C. Pidacks, Science 109 199 (1949). (Isolation) 

Benjamin M. Duggar, U. S. Patent 2,482,055 (1949). 

C. R. Stephens, L. H. Conover, F. A. Hochstein, P. P. Regna, 
F. J. Pilgrim, K. J. Brunings and R. B. Woodward, /. Am. 
Chem. Soc. 74 4976 (1952). 



Pfizer Handbook of Microbial Metabolites 280 

C. W. Waller, B. L. Hutchings, R. W. Broschard, A. A. Gold- 
man, W. J. Stein, C. F. Wolf and J. H. Williams, ibid. 74 4981 
(1952). 

609 Bromotetracycline, CooHogOgN.Br, m.p. 170-172°, [ale'" -196° 
(in 0.1 N hydrochloric acid). 




000 
H H 



CONH2 



Streptomyces aureofaciens 

P. Sensi, G. A. DeFerrari, G. G. Gallo and G. Holland, II 
Farmaco Ed. sci. (Pavia) 10 337 (1955). 

610 Oxytetracycline ( Terramycin ) , C00H04O9N2, light-yellow crys- 
tals, m.p. (anhydride) '--'185° (dec), [ajo'^ (dihydrate) 
-196.6° (c 1.0 in 0.1 N hydrochloric acid). 

CH3 CH3 

CH3 OH OH N 

OH 




CONH2 
OH O OH ^ 

Streptomyces rimosus 

A. C. Finlay, G. L. Hobby, S. Y. P'An, P. P. Regna, J. B. 
Routien,-D. B. Seeley, G. M. ShuU, B. A. Sobin, I. A. Solomons, 
J. W. Vinson and J. H. Kane, Science 111 85 (1950). (Iso- 
lation) 

Ben A. Sobin, Alexander C. Finlay and Jasper H. Kane, 
U. S. Patent 2,516,080 (1950). 

Peter P. Regna, I. A. Solomons, Kotaro Murai, Albert E. 
Timreck, Karl J. Brunings and W. A. Lazier, J. Am. Chem. 
Soc. 73 4211 (1951). 

C. R. Stephens, L. H. Conover, F. A. Hochstein, P. P. Regna, 
F. J. Pilgrim, K. J. Brunings and R. B. Woodward, ibid. 74 
4976 (1952). 

F. A. Hochstein, C. R. Stephens, L. H. Conover, P. P. Regna, 
R. Pasternack, P. N. Gordon, F. J. Pilgrim, K. J. Brunings and 
R. B. Woodward, ibid. 75 5455 (1953). (Structure) 

611 Antibiotic X-340, CosHooOe, yellow needles, m.p. 330° (dec). 
An antibiotic isolated from the mycelium of an uniden- 



28l 



Tetracycline, Analogues and Related Substances 



tified streptomycete. The molecular weight was about 
390. Contained 3 — OH groups (one acidic) and one 
C — CH;5 group. Monomethyl derivative with diazometh- 
ane. Mono- and tri-acetates were formed, depending on 
method. The infrared absorption pattern was similar to 
that of Terramycin. The following partial structure was 
proposed : 

— CH3 
—OH 
— CH=CH 

I — 2H 
— CH=CH 
2 double bonds 
_1 unplaced O-atom 

V. C. Vora, K. Shete and M. M. Dhar, J. Sci. Ind. Research 
(India) 16C 182 (1957). (Isolation) 

612 2- Ace tyl-2-decarboxamidooxy tetracycline ( Terramycin-X ) (Hy- 
drochloride), CogHsgOgN-HCl, ycllow crystals, m.p. 201- 
203°, [ale'" -46.6° (c 0.9 in 0.1 N hydrochloric acid). 





Streptomyces rimosus 

F. A. Hochstein, M. Schach von Wittenau, F. W. Tanner, Jr. 
and K. Murai, /. Am. Chem. Soc. 82 (1960). (In press) 

613 Tetracycline (Achromycin, Tetracyn, Polycycline, Panmycin), 
C22H24OSN0, yellow crystals, m.p. 170-175° (dec), [aW^ 
-239° (c i.O in methanol). 




CONH2 



Streptomyces sp. 



Pfizer Handbook of Microbial Metabolites 



282 



Tetracycline was first prepared by catalytic dechlorina- 
tion of chlortetracycline but was later isolated as a pri- 
mary fermentation product. 

P. Paul Miuieri, Melvin C. Firman, A. G. Mistretta, Anthony 
Abbey, Clark E. Bricker, Neil E. Rigler and Herman Sokol, 
"Antibiotics Annual 1953-1954," Medical Encyclopedia, Inc., 
New York, p. 81. (Isolation) 

614 Quinocyclines (PA-121) 

A complex of tetracyclic amphoteric antibiotic yellow 
pigments, which in some respects resemble nitrogen-con- 
taining hydroxy anthraquinones. 

Six active components have been separated and analy- 
ses and color reactions were determined. 

Two components have an aglycone with the probable 
empirical formula Co-H^oOc.N^. 

Streptomyces sp. 

W. D. Celmer, K. Murai, K. V. Rao, F. W. Tanner, Jr. and 
W. S. Marsh, "Antibiotics Annual 1957-1958," Medical En- 
cyclopedia, Inc., New York, p. 484. (Isolation) 

Charles R. Stephens, unpublished. (Empirical formula) 

615 7;-Pyrromycin, C30H35O11N (Hydrochloride), red crystals, m.p. 

162° (dec), [a]c,r" +132 ±27° (c 0.4 in methanol). 



CH3I 




>Rhodosamine 



OH O 



CH2 
COOCH3 



A streptomycete 

The relationship to e-pyrromycinone and to the cineru- 
bins should be noted. 

Hans Brockmann and Werner Lenk, Chem. Ber. 92 1904 
(1959). (Structure) 

Hans Brockmann and Hans Brockmann, Jr., Naturwis- 
senschaften 47 135 (1960). 



283 Tetracycline, Analogues and Related Substances 

616 Aklavin, C:^oH;i-0,,N (Hydrochloride) orange crystals, m.p. 

197°. 

Amphoteric. Contains an amino sugar, C.^Hj^OiN, iso- 
meric with mycaminose or amosamine linked glycosidi- 
cally to the secondary hydroxyl group. 

Streptomyces sp. 

F. Strehtz, H. Flon, U. Weiss and I. N. Asheshor, ]. Bacte- 
riol. 72 90 (1956). 

617 Cinerubins 

Cinerubins A and B are isomeric red bases, with the 
empirical formula C44H.-,.,OisN±CHo. The chromophoric 
aglycone has been shown to be identical with e-pyrromy- 
cinone. Both cinerubins also contain three sugars, two 
of these being the same in both compounds, but the third 
one being characteristic. The structures of these sugars 
have not been reported yet. 

Streptomyces antihioticus (Waksman and Woodruff) 
Waksman et Henrici, S. galiloeus Etthnger et al., S. niveo- 
ruber Ettlinger et al. 

Leopold Ettlinger, Ernest Gaumann, Ralf Hiitter, Walter 
Keller-Schierlein, Frlederich Kradolfer, Lucien Neipp, Vlado 
Prelog, Pierre Reusser and Hans Zahner, Chem. Ber. 92 1867 
(1959). 



14 



Aromatic Compounds Not Classified 
Elsewhere 



This chapter includes a heterogeneous group of aromatic 
compounds which arise from different biosynthetic routes. Cin- 
namic acid and its derivatives undoubtedly are formed by way 
of the shikimic acid pathway.^' ^ The occurrence of anisalde- 
hyde and anisic acid derivatives in the same fermentation with 
methyl p-methoxycinnamate suggests that the former may be 
degradation products of the latter. 

Chloramphenicol, too, has a Ce-Ca skeleton which seems to 
relate it to the shikimic acid pathway. It has been shown that 
p-nitrophenylserinol does not act as a precursor, and, when it is 
added to fermentations, it is acetylated but not dichloroacety- 
lated. C"-Labeled p-nitrophenylserinol is not incorporated into 
the chloramphenicol molecule nor is C"-labeled dichloroacetic 
acid. Thus, what appears to be a logical step in the biosynthe- 
sis — the dichloroacetylation of p-nitrophenylserinol — does not 
occur.^ 

The tricarboxylic acid produced by Chaetomium indicum is 
evidently formed by the condensation of a-ketoglutaric acid with 
phenylpyruvic acid. 

The lichen acids of this chapter show a provocative symme- 
try, and the incorporation of amino acids into two of them is in- 
teresting. The diphenylbutadiene structure has been found 
also in xanthocillin. Apparently there has been no experimen- 
tal study of their biogenesis. 

^ Friedrich Weygand and Heinz Wendt, Z. Natiirforsch. 14b 421 
(1959). 

2T. A. Geissman and T. Swain, Chem. and Ind., 984 (1957). 

3 David Gottlieb, P. W. Robbins and H. E. Carter, J. Bacterial. 72 
153 (1956). 



285 Aromatic Compounds Not Classified Elsewhere 

618 Benzoic Acid, C7H,;0o, colorless tablets, m.p. 122.5°. 

COOH 



Yeast 

Richard Kuhn and Klaus Schwarz, Ber. 74 1617 (1941). 

619 Anisaldehyde, CsHgOo, oily liquid, b.p. 248°, n^'^ 1.5764. 

CHO 



OCH3 

Trametes suavolens (Linn.) Fr., Lentinus lepideus, 
Daedalea juniperina 

J. H. Birkinshaw, A. Bracken and W. P. K. Findlay, 
Biochem. }. 38 131 (1944). 

J. H. Birkinshaw and P. Chaplen, ibid. 60 255 (1955). 

620 trans-Cinnamic Acid, CgHgOs, colorless crystals, m.p. 133°. 

/ ^CH=CH— COOH 

Ceratostomella firnbriata (on sweet potato culture) 
Takashi Kubota and Keizo Naya, Chem. and Ind., 1427 
(1954). 

621 trans-Cinnamic Acid Amide, C9H9ON, colorless crystals, m.p. 

147-149°. 

/ '^CH=CH— CONH2 

Streptomyces sp. 

Yasuharu Sekizawa, /. Biochem. Japan 45 9 (1958). (Iso- 
lation ) 

622 Methyl Anisate, C9H10O3, colorless crystals, m.p. 48°, b.p. 256°. 

COOCH3 



OCH3 



Pfizer Handbook of Microbial Metabolites 286 

Travietes suavolens (Linn.) Fr., Lentinus lepideus 
J. H. Birkinshaw, A. Bracken and W. P. K. Findlay, Biochem. 
J. 38 131 (1944). 

623 Methyl trans-Cinnamate, CioHk.Oo, clear, pale yellow oil, b.p. 
94-110° (2-3 mm.) or white crystals, m.p. 35-37°, nn'^ 
1.5766 



rv< 



CH=CH— COOCH3 

Lentinus lepideus Fr. (artificial medium) 
John Howard Birkinshaw and Waher Phihp Kennedy Find- 
lay, Biochem. J. 34 82 (1940). 

624 Methyl p-Coumarate, CioHjoOg, colorless crystals, m.p. 137- 

139°. 

HO—/ \— CH=CH— COOCH3 

Lentinus lepideus 

H. Shimazono and F. F. Nord, Arch. Biochem. and Biophys. 
78 263 (1958). 

625 Methyl p-Methoxycinnamate, CnHioOy, colorless crystals, m.p. 

88°. 

CH3O— / y-CH=CH— COOCH3 

Lentinus lepideus Fr. (artificial medium) 
John Howard Birkinshaw and Waker PhiHp Kennedy Find- 
lay, Biochem. J. 34 82 (1940). 

626 Chloramphenicol (Chloromycetin, Levomycetin),CiiHi20.-,NoCl2, 

colorless crystals, m.p. 149.7°, [oc]d-'' —25.5° (in ethyl ace- 
tate). 

OH 

O2N— <^^>— CH— CH— CHoOH 

NHCOCHCI. 

Streptomyces venezuelae 

John Ehrlich, Quentin R. Bartz, Robert M. Smith, Dwight A. 
Joslyn and Paul R. Burkholder, Science 106 417 (1947). (Iso- 
lation) 

John Controulis, Mildred C. Rebstock and Harry M. Crooks, 
Jr., J. Am. Chem. Soc. 71 2463 (1949). (Synthesis) 



287 Aromatic Compounds Not Classified Elsewhere 

Loren M. Long and H. D. Troutman, ibid. 71 2469 (1949). 

627 1,8-Dimethoxynaphthalene, C,^.H,oO.>, colorless crystals, m.p. 
158-161°. 

CH3O OCH;, 



Daldinia concentrica 

8-Methoxyl-l-naphthol also was identified (by paper 
chromatography ) . 

D. C. Allport and J. D. Bu'Lock, /. Chem. Soc, 654 (1960). 

628 4-Carboxy-2-oxo-3-phenylhept-3-enedioic Acid, C14H12O7, colorless 
prisms, m.p. 170° (dec). 

O COOH 

HOOC— C— C=C— CH.— CH.— COOH 



Chaetomium indicum Corda 

The yield was 250-500 mg. per liter. In addition to 
the acid above, two uncharacterized compounds were iso- 
lated in smaller quantities: Metabolite A, C2,jH:^70(;N, pale 
yellow needles, m.p. 159°, [a],r" +11.4° (c 1.022 in chlo- 
roform). Yield 1.5-2.0 g. from 100 1. of broth. Soluble 
in aqueous NaHCOg. Wine red FeCl, test. Formed an 
insoluble green-blue copper derivative. 

Metabolite B, colorless prisms, m.p. 146°, [a],,-" +120° 
(c 1.01 in chloroform). 

Analysis: C 68.1, H 8.2, N 2.7, C-methyl 12^^. Same 
color tests as A above. 

D. H. Johnson, Alexander Robertson and W. B. Whalley, /. 
Chem. Soc, 2429 (1953). 

629 Pulvic Anhydride, CiyHi„04, yellow needles, m.p. 222-224°. 

o c=o 



o=c- 

Sticta aurata Ach. 



Pfizer Handbook of Microbial Metabolites 288 

O. Hesse, J. prakt. Chem. 170 334 (1900). 

630 Calycin, CisHjoOg, orange-red crystals, m.p. 244°. 

o c=o 

^-cJ-c=c-Q) 
\ c — -o 

OH,| 

o 

Lepraria candelaris Schaer., Sticta aurata Ach. and 
Sticta crocata Ach. 

Mitizo Asano and Yukio Kameda, Ber. 68 1568 (1935). 

631 Vulpinic Acid, C19H14O5, yellow crystals, m.p. 148°. 

COOCH3 

0-kc-c=c-Q) 

I OH I 

o c=o 

Evernia vul-pina L., Cyphelium chrysocephalum Ach., 
Calicium chlorinum Korper, Cetraria juniperina Fr. var. 
tubulosa Schaer and Cetraria pinastri (Scop.) 

P. Karrer, K. A. Gehrckens and W. Heuss, Helv. Chim. Acta 
9 446 (1926). (Structure) 

632 Pinastric Acid ( Chry socetraric Acid), CooHigOe, orange needles, 

m.p. 200-203°. 

OH COOCH3 

_ CH3O— /^^c=c— c=c— ^^> 
o=c o 

Lepraria fiava (Schreber. ) f. quercina, Cetraria pinastri 
(Scop.), Cetraria tubulosa (Schreb.), Cetraria juniperina 
L. (Ach.) 

Mitizo Asano and Yukio Kameda, Ber. 68 1565 (1935). 
(Structure) 

633 Leprapic Acid (Leprapinic Acid, Methyl 2-Methoxypulvlnate ) , 

CsoHieOg, golden plates, m.p. 159°. 

COOCH3 OH 

\^ o — c=o 
OCH3 



289 Aromatic Compounds Not Classified Elsewhere 

Lepraria citrina 

O. P. Mittal and T. R. Seshadri, J. Chem. Soc, 3053 (1955). 
(Isolation) 

Idem., ibid., 1734 (1956). (Synthesis) 

634 Mycolutein, C00H24O6N (proposed), bright yellow tablets, m.p. 

157°, [afn"' +54° (c 1 in chloroform). 

Contains an aromatic nucleus. Alkali-unstable. Neg- 
ative FeCla. Decolorizes bromine with HBr evolution. 
Streptomijces sp. 

Henry Schmitz and Robert Woodside, Antibiotics and 
Chemotherapy 5 652 (1955). 

635 Epanorin, CosHojOoN, yellow needles, m.p. 135°, [ajo^^ -1.86 

±0.2° (c 6.48 in chloroform). 

CH2— CH(CH3)2 
I 
NH— CH— COOCH3 

I 

C=0 OH 

<Q>-C=C-C=C-Q) 

o c=o 

Lecanora epanora Ach. 
Zeorin was found in the same extract. 
Robert L. Frank, S. Mark Cohen and James N. Coker, /. Am. 
Chem. Soc. 72 4454 (1950). (Structure and synthesis) 

636 Rhizocarpic Acid, CogHssOeN, yellow needles, m.p. 177°, [ajn^" 

+ 110.4° ±2.1° (c 1.22 in chloroform). 

COOCH3 

NH— CH— CH2— (^^> 
C=0 OH 

<Q_kc-c=c-Q> 
o c=o 

Rhizocarpon geographicum L.,R. viridiatrum Flk., Cali- 
cium hyperellum Ach. 

Robert L. Frank, S. Mark Cohen and James N. Coker, /. Am. 
Chem. Soc. 72 4454 (1950). (Synthesis) 



15 



Amines 



Much remains to be learned concerning the earlier stages of 
nitrogen metabolism in microorganisms. Practically, the abil- 
ity of certain soil bacteria (in combination with legumes) to fix 
gaseous nitrogen has been exploited for many years. Research 
in this area has been reviewed.^ Ammonia, methane, hydrogen 
and water probably were present in the atmosphere of the primi- 
tive earth, and it has been shown- that amino acids can be 
formed by electric discharges through such mixtures. 

While we are primarily concerned in this compilation with 
metabolites isolated from microorganisms growing in the wild 
state or cultivated on an essentially glucose medium, the more 
complex amines are generally only remotely derived from sugar, 
often by way of the amino acids. A large hterature exists on 
the ability of bacteria to decarboxylate amino acids to amines, 
these experiments generally involving addition of the amino 
acid to the medium. It has been shown,^ however, that many 
bacteria which produce amines on a casein hydrolysate medium 
do not do so on a synthetic medium with ammonium salts the 
only nitrogen source. Studies with Escherichia coli* indicate 
that aspartic acid and alanine and perhaps glutamic acid serve 
as important nitrogen entry vehicles. These acids can supply 
the total nitrogen requirement if no ammonium ion is available, 

^ William D. McElroy and Bentley Glass, "Inorganic Nitrogen 
Metabolism," Johns Hopkins Press, Baltimore, 1956. 

2 Stanley L. Miller, Science 117 528 (1953); idem., J. Am. Chem. 
Soc. 77 2351 (1955). 

3 H. Proom and A. J. Woiwod, J. Gen. Microbiol. 5 930 (1951). 

* "Studies of Biosynthesis in E. coli," Carnegie Institute of Wash- 
ington Publication 607, Washington, 1955. 



291 Amines 

and, even when it is, much of the cellular nitrogen is derived 
from them by transamination. 

Within the frame of our present endeavor there seems to have 
been little systematic, comparative study of the amine metab- 
olites of microorganisms. This has been true particularly of 
the fungi, which generally have been considered to have a 
poorer nitrogen metabolism than the bacteria. Apparently this 
situation is being remedied, at least for higher fungi. Recently 
the amine content of 105 species, representing 18 families of 
higher fungi, was investigated.^ It was found that ammonia 
was distributed universally, and that the ammonia content in- 
creased with the age of the fruiting body. Methylamine oc- 
curred in 22 species, diinethylamine in 10, trimethylamine in 8, 
isoamylamine in 19 and /^-phenylethylamine in 4. Earlier work 
was reviewed also, a distinction being made between the amines 
present in fresh fruiting bodies and those present after au- 
tolysis. 

Also an exceptionally thorough analysis was made recently 
of the basic constituents of the fruiting body of a single basidio- 
mycete, Polyporus sulfureusJ' These included amines, basic 
amino acids, nucleotides and betaines. Many of the simple 
amines produced by Claviceps purpurea have been identified 
during the extensive studies of ergot, and these are listed in the 
introduction to the section on ergot alkaloids in the chapter on 
Heterocycles. 

Muscarine, a compound which might have been classified un- 
der several different chapter headings, is apparently a deriva- 
tive of a l-amino-3,6-desoxyhexose and is probably more di- 
rectly connected with sugar metabolism than many of the 
amines listed here. 

Amino sugars and other complex amines are listed elsewhere 
under more appropriate classifications. 

It has been shown that putrescine furnishes the 4-carbon 
atom moiety of spermine and spermidine in Neurospora crassaj 
and that methionine supplies the 3-carbon chain of spermidine 
in the same organism."^ It is known that ATP and Mg"* are re- 

^Elard Stein von Kamienski, Planta 50 331 (1958). 
6 P. H. List, Planta Med. 6 424 (1958). 

^ H. Tabor, S. M. Rosenthal and C. W. Tabor, Federation Proc. 15 
367 (1956). 

8 Ronald C. Greene, /. Am. Chem. Soc. 79 3929 (1957). 



Pfizer Handbook of Microbial Metabolites 



292 



quired. A mechanism such as the one shown here (abbreviated) 





11'^+ HiNlCHilaNHlCHshNHi 



Spermidine 



CH3— S— CH2 



OH OH 



CH— COOH 

I 
NH2 

S-Methyl-S-adenosyl- 
methionine 



toH 



OH OH 



may be operative. 

637 Ammonia, NH3, colorless gas. 

NH3 

Widely distributed in the fruiting bodies of the higher 
fungi and lichens. The content increases with age. 
Elard Stein von Kamienski, Planta 50 331 (1958). 

638 Methylamine, CH5N, colorless gas. 

CH3NH2 

Russula (11 spp.), Lactarius deliciosus, L. vellereus, 
L. helvus. Boletus edulis, B. appendiculatus, Scleroderma 
vulgare-; Anthurus muellerianus, Mutinus caninus, Tra- 
chypus versipellis, Dermocybe (Cortinarius) cinnamo- 
mea, Lepiota clypeolaria, Pholiota mutabilis, Sticta fulig- 
inosa, S. sylvatica, Polyporus sulfureus 

Elard Stein von Kamienski, Planta 50 331 (1958). 

P. H. List, Planta Med. 6 424 (1958). 

639 Ethylamine, C2H7N, volatile liquid, b.p. 16.6°. 

CH3CH2NH2 

Claviceps purpurea, Polyporus sidfureus 
Maximilian Steiner and Elard Stein von Kamienski, Natur- 
wissenschaften 42 345 (1955). 

P. H. List, Planta Med. 6 424 (1958). 



293 Amines 

640 Dimethylamine, CoH^N, colorless gas, b.p. 7°. 

CHs— NH— CH3 

Phallus impudicus, Clathriis ruber, Russula aurata 

Gustav Klein and Max Steiner, Jahrb. wiss. Bot. 68 602 
(1928). 

R. sardonia, R. turci, R. lepida, R. cyanoxantha, R. 
grisea, R. olivacea, R. vesca, R. alutacea, Sticta sylvatica, 
Polyporus sulfureus 

Elard Stein von Kamienski, Planta 50 333 (1958). 

P. H. List, Planta Med. 6 424 (1958). 

641 Ethanolamine, C2H7ON, colorless oil, b.p. 171°, Hd^" 1.4539. 

HOCH0CH0NH2 

Neurospora crassa (and probably in) Boletus edulis, 
B. versipellis, Xerocomus badius, Lepiota clypeoloris, 
Pholiota mutabilis, Tricholoma nudum, Russula macu- 
lata, R. turci, Lactarius vellercus, Amanita muscaria, 
Polyporus sulfureus 

George L. Ellman and Herschel K. Mitchell, /. Am. Chem. 
Soc. 76 4028 (1954). 

Elard Stein von Kamienski, Planta 50 331 (1958). 

P. H. List, Planta Med. 6 424 (1958). 

642 Aminoacetone, C3H7ON, colorless crystals, m.p. 130.5°. 

CH3COCH2NH2 

Staphylococcus aureus 

W. H. Elliot, Nature 183 1051 (1959). 

643 Trimethylamine, C3H9N, colorless gas (fishy odor), b.p. 3°. 

CH3 CH3 

CH3 

Boletus edulis, Ustilago maydis. Phallus impudicus, 
Claviceps purpurea, Tilletia laevis, T. tritici, Clathrus 
ruber, Russula spp. Sticta spp. 

J. Zellner, Monatsh. 31 617 (1910). 

William Fielding Hanna, Hubert Bradford Vickery and 
George W. Pucher, /. Biol. Chem. 97 351 (1932). (Isolation) 

Maximilian Steiner and Elard Stein von Kamienski, Natur- 
wissenschaften 42 345 (1955). 



Pfizer Handbook of Microbial Metabolites 294 

644 n-Propylamine, C3H9N, liquid, b.p. 50°. 

CH3CH2CH2NH2 

Claviceps piirpiirea, Polyporiis sulfiirens 
Maximilian Steiner and Elard Stein von Kamienski, Natur- 
wissenschaften 42 345 (1955). 

P. H. List, PZanta Med. 6 424 (1958). 

645 Isopropylamine, C;{H,)N, liquid, b.p. 33°. 

CH3— CH— CHs 

NH2 

Claviceps purpurea 

Maximilian Steiner and Elard Stein von Kamienski, Natur- 
wissenschaften 42 345 (1955). 

646 Methylaminoethanol, C.iHgON, slightly viscous liquid, b.p. 159°. 

HOCH2CH2NHCH.S 

Neurospora crassa mutant 

N. H. Horowdtz, J. Biol. Chem. 162 413 (1946). 

647 n-Hexylamine, C6H,-,N, liquid, b.p. 129°. 

CH3(CH2)5NH2 

Claviceps purpurea 

Maximilian Steiner and Elard Stein von Kamienski, Natur- 
wissenschaften 42 345 (1955). 

648 Isobutylamine, C4H11N, liquid, b.p. 68°. 

CH3 

\ 

CH— CHo— NHo 

/ 
CH3 

Claviceps purpurea 

Maximilian Steiner and Elard Stein von Kamienski, Natur- 
wissenschaften 42 345 (1955). 

649 l-Amino-2-methyl-2-propanol, C,Hi,ON, liquid, b.p. 151°. 

CH3 

H2N— CH2— C— CH3 

OH 
Neurospora crassa 



^95 Amines 

George L. Ellman and Herschel K. Mitchell, 7. Am. Chem. 
Soc. 76 4028 (1954). 

650 Putrescine, C4H10N0, crystals, m.p. 27°. 

H2NCH.CH,CHoCH,NHo 

Boletus edulis, B. luteus, B. elegans, Amanita muscaria 
C. Reuter, Z. physiol. Chem. 78 167, 223 (1912). 
Albert Kiing, ibid. 91 241 (1914). 

Werner Keil and Hans Bartmann, Biochem. Z. 280 58 
(1935). 

651 Histamine, C.-^H^Nj^, deliquescent needles, m.p. 83° (Hydrochlo- 

ride) m.p. 244-246° (Picrate) m.p. 160°. 

r^^— CH2CH0NH2 



Nv. .NH 



Claviceps purpurea, Coprinus comatis Gray 
Paul Heinz List, Arch. Pharm. 291 502 (1958). 

652 Isoamylamine, C-,Hi;jN, liquid, b.p. 95-97°. 

CH3 

\ 

CH— CH2— CH2— NH2 

/ 
CH3 

Boletus edulis, B. sanguineus, B. queletii, B. luridus, 
B. regius, B. appendiculatus. Phallus impudicus, Claviceps 
purpurea, Amanita phalloides, Marasmium peronatus, 
Russula foetens, R. turei, R. maulata, Trachypus scaber, 
Xeroeomus sanguineus, X. suhtomentosiis , Mutinus cani- 
nus, Lycoperdon piriforme, L. gemmatum, Phlegmacium 
mellioleus, Nematoloma fasciculare, Polyporus sulfureus 

Maximilian Steiner and Elard Stein von Kamienski, Natur- 
wissenschaften 40 483 (1953). 

Elard Stein von Kamienski, Planta 50 334 (1958). 

P. H. List, Planta Med. 6 424 (1958). 

653 Dimethylhistamine, C-H,;^N;j ( Dihydrochloride ) m.p. 245-250° 

(dec). 

CH3 

/ 
1^^— CH2CH2N 

N^ NH CH3 



Pfizer Handbook of Microbial Metabolites 296 

Coprinus comatis Gray 

Paul Heinz List, Arch. Pharm. 291 502 (1958). 

654 Acetylcholine, C7H17O3N, colorless, hygroscopic powder. 

(CHslsN— CH2CH2OCOCH3 

® 

Streptobacteriuvi plantarum 

Acetylcholine is produced also by the ergot fungus, 
Claviceps purpurea, and probably by many other micro- 
organisms. 

Yield: about 160 y per milUUter from the first organ- 
ism above. 

Adolf Wacker, Adolf Roth, Heinz Sucker and Otto Dann, 
Ann. 601 202 (1957). 

655 Spermidine, C7H19N3, unstable oil, b.p. 128° (14 mm.). 

H2N(CH2)3NH(CH2)4NH2 

Yeast, Neurospora crassa 
Occurs as the phosphate. 

H. Tabor, S. M. Rosenthal and C. W. Tabor, Federation 
Proc. 15 367 (1956). 

Ronald C. Greene, J. Am. Chem. Soc. 79 3929 (1957), 

656 ^-Phenylethylamine, CgHuN, liquid, b.p. 196-198°. 

/ V-CHsCHzNHz 

Boletus edulis, B. luteus, Claviceps purpurea, Polyporus 
sulfureurS, Marasmius peronatus, Phlegmaciuni melliolus, 
Nematoloma fasciculare, Pholiota mutabilis 

C. Reuter, Z. physiol. Chem. 78 167 (1912). 

Werner Keil and Hans Bartmann, Biochem. Z. 280 58 
(1935). 

Elard Stein von Kamienski, Planta 50 335 (1958). 

P. H. List, Planta Med. 6 424 (1958). 

657 Tyramine, CsHj^ON, colorless crystals, m.p. 164°. (Picrate), 

m.p. 206°. 

HO—/ V-CH2CH,,NH2 



Coprinus comatis Gray, Claviceps purpurea 
Paul Heinz List, Arch. Pharm. 291 502 (1958). 



297 Amines 

658 Muscarine, CgHigOoN, white crystals, Hydrochloride [a]u^° 
+ 1.57° (in water). 




Amanita muscaria 

F. Kogl, C. A. Salemink, H. Schouten and F. Jellinck, Rec. 
trav. chim. 76 109 (1957). (Structure) 

E. Hardegger and F. Lohse, Helv. Chim. Acta 40 2383 
(1957). (Synthesis and configuration) 

P. J. Fraser, Brit. J. Pharmacol. 12 47 (1957). (Pharma- 
cology) 

659 Muscaridine, CgHooOoNCl, isolated as the chloroaurate, 

CgHooAuCl^O.N, " m.p. 129-131° (dec), [aW +20.5° 
±0.5° (c 8.3 in water). 

CI® CH3 

© I 
CH3— N— CH2— CH2— CHo— CH— CH— CH3 

I I I 

CH3 OH OH 

Amanita muscaria L. 

F. Kogl, C. A. Salemink and P. L. SchuUer, Rec. trav. 
chim. 79 485 (1960). (Isolation) 

C. A. Salemink and P. L. SchuUer, ibid. 79 278 (1960). 
(Synthesis) 

660 Spermine, C10H26N4, deliquescent, C02-absorbing crystals. 

Phosphate: m.p. 230-234° (dec). 

H2N(CH.2)3NH(CH2)4NH(CH2)3NH2 

Yeast, Neurospora crassa 
Occurs phosphorylated. 

H. Tabor, S. M. Rosenthal and C. W. Tabor, Federation 
Proc. 15 367 (1956). 

Ronald C. Greene, J. Am. Chem. Soc. 79 3929 (1957). 

661 Bufotenin, CioHigONo, colorless crystals, m.p. 146°. 

HO 

-CH2CH2N(CH3)2 



Oj 



H 



Pfizer Handbook of Microbial Metabolites 298 

Amanita mappa and certain related species 
Bufotenin occurs also in the skin secretions of toads. 
Theodor Wieland and Werner Motzel, Ann. 581 10 (1953). 

662 Necrosamine, C..,)H44N2 (Hydrochloride) crystals, m.p. ~275° 
(dec). 

CH3— (CHoJu— CH— CH— CH2— CH2— CH3 

NH2 NH2 

Escherichia coli 

This amine was a component of the phospholipide frac- 
tion. 

Miyoshi Ikawa, J. B. Koepfli, S. G. Mudd and Carl Niemann, 
J. Am. Chem. Soc. 75 3439 (1953). 



16 



Amino Acids and Related Compounds 



A general review of the intermediary metabolism of amino 
acids would be disproportionate to the scope of this book. It is 
only possible to sketch in here some relationships and biosyn- 
thetic sequences which may serve as reminders or as guides for 
the novice. 

As in acetate metabolism microorganisms have been used to 
explore the network of metabolic relationships among the amino 
acids. Many of these have proved quite general, yet it is only 
necessary to consider the unusual amino acids which have been 
isolated from microbial sources to realize the differences from 
human metabolism. 

In this section principally free amino acids are considered. 
Polypeptides are listed and discussed in the succeeding section. 
Amino acid isolation and assay formerly were tedious and gen- 
erally confined to analysis of hydrolysates of total proteins. Pa- 
per chromatography and reliable microbiological assays have 
made possible the separation and assay of the low concentra- 
tions of amino acids evolved into fermentation broths. 

The older work on fungi has been reviewed.^ A semiquanti- 
tative survey of the free amino acids of a taxonomic range of 
fungi gave the results shown in Table P on page 300. In 
general there were found no outstanding differences in the 
quantities or types of amino acids produced by the different 
fungi, nor in the types produced by fungi as compared with 
those of higher plants. The absence of tryptophan in all species 
examined is noteworthy. Four unidentified compounds were 
found in various fungi. These were suggested tentatively as 

1 Jackson W. Foster, "Chemical Activities of Fungi," Academic 
Press, New York, 1949. 

2R. Close, Nature 185 609 (1960). 



P P: c -^ 



— a 

UJ ^ 

< j: 



^1 


+ + + + 


+ + + 




+ +CN^ 


£ a 


+++ 1 1 +++ 1 


1 + + + 


1 + + + 


1 H — h -^' <> o 


CO 3 

a 


+ + + 






+ + 


i^ 


+ + + 


+ 




+ +CSO. 


D 1 


++++++++ 1 


+ + + 


1 ++ 1 


+ + + '*<)->»• 


if 


+++ +++ 


+ + 


+ + 


+ + 






+ 






IP 


+++ +++ 


+ 


+ + 


+ + +^r. 


s 1 

'11 


+++ 1 ++++ 1 


1 ++ 1 


1 + + + + + + ;; K^ 1 


+ + + + 


+ 




+ 


■omo- 

rea 

lulosa 


+++ +++ 


+ + 


+ + 


+ + +CN- 


+++ 1 ++++ 1 


+++++++ 1 


+ + +W «i^ 


5 "i 


+ + + 


+ 


+ 


+ + 


Thorn - 
nidium 
elegans 


+ + + + + 






+ +CNO 


+++++++ 1 + 
+ + + 


1 ++ 1 


+ + + 1 


+ + + '*' CO «0 

+ + 


• ^ + 

O u 


+++ ++++ 


+ + 


+ + 


+ + CNK 


1-g 


++++++++ 1 


+ + + 1 


+ + + 1 


1 + + ->t <d <o 


a. E .^ 


+ + + 


+ 




+ + 


■ S 1 
o i; ? 


+++++++++ 


+ + + 


+ + 


+ +^. 


>.. j: o 


+++++++++++++ 


1 + + + + + + ;; ^ o. 1 


■^ a 8 


+ + + + 


+ 




+ + 


E E "^ 

3 3 O 


+++ + ++ 


+ + + 


+ + 


+ + + ^<n 


:£ E Q 


+++ 1 ++++ 1 


1 + + + 


1 ++ 1 


+ + + CO uS o 


a. 3 ^ 


+ 


+ 




+ 




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II 


>• 




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fe "o 






^1 1^ 

" u 4> «> J° 4> 
u .= c c a, c 


0) 4) 4> 


£ c « 
° a .S «> 


E 

:2 41 ■■= 

U C 4) c 

5) .£ X 3- 




t = ■£ 'E .E (u c c «> 


c c C 


c "0 £ •- 


i! 5 p - ° 




<0 acQ.?-<oOf£a: 


2 o) ti •:; 


5J = 4) £ 


u < O ^ it ^ 



i; 



30I 



Amino Acids and Related Compounds 



a-aminoadipic acid, 3,4-dihydroxyphenylalanine, ethanolamine 
and taurine. 

The amino acids of some algae have been reported,^ and also 
those of Fusariinn lycopersici:' A quantitative study was made 
of the amino acid composition of Usiilago maydis fermentation 
broth.' Of the 3.5 mg. per milliliter of NH4" nitrogen added, 
2.9 mg. per milliliter remained extracellular. This extracellular 
nitrogen contained 1.17 mg. per milliliter of organic nitrogen 
and 1.74 mg. of residual NH4* nitrogen. 



TABLE II 
Amino Add Composifion of Usfilago maydis Fermenfafion Broth 



Amino acid* 


Unhydrolyzed broth 


Hydrolyzed broth 


/xgm /ml. 


jugm.N/ml. 


^igm./ml.f 


Atgm.N/ml.f 


Lysine 


387 
997 
155 
200 
894 
200 

290 
387 
276 
276 

263 
267 

139 
40 
65 


64.2 
320.5 
42.0 
21.0 
85.1 
38.1 

25.7 
41.3 
29.5 
30.1 

32.0 

22.6 

10.7 

5.6 

6.0 


413 
1136 
182 
506 
945 
295 
406 
279 
368 
212 
307 
237 
289 
389 
383 


79 2 


Arginine 

Histidine 

Aspartic acid . . . 
Glutamic acid. . . 

Glycine 

Alanine! 

Valine 


365.3 
49.3 
53.2 
90.0 
55.0 
63.8 
33 4 


Leucine 

Isoleucine 


39.3 
22.7 
40.9 


Threoninel 


27.9 
35.2 


Phenylalanine. . . 

Tyrosine 

Tryptophan 

Methionine 


33.0 
29.6 




4.79 mgm. 


0.774 mgm. 


6.35 mgm. 


1.02 mgm. 



* The amino acids in the hydrolyzed broth and the bosic amino acids in the unhydrolyzed broth were 
separated chromatographically and assayed colorimetrlcally. The other amino acids in the unhydrolyzed 
broth were assayed microbially. 

t Values expressed as ^gm. per milliliter in terms of original broth. 

t Valid microbial assays were not obtained. 

3L. Fowden, ibid. 167 1030 (1951); Borje Wickberg, Acta Chem. 
Scand. II 506 (1957). 

4V. Fliick and K. H. Richie, Phytopath. Z. 24 455 (1955). 

5 Eugene L. Dulaney, E. Bilinski and W. B. McConnell, Can. J. 
Biochem. and Physiol. 34 1195 (1956). 



Pfizer Handbook of Microbial Metabolites 



302 



In this case the broth was hydrolyzed and compared with the 
original to eUminate interference by small peptides in the mi- 
crobial assays. Tryptophan and methionine were destroyed by 
the hydrolysis and chromatography procedure and are absent 
from the second part of the table. It was found that 53% of 
the extracellular organic nitrogen was represented by free amino 
acids. Some strains of Ustilago maydis produce 200—300 jxg. 
of lysine per milliliter." 

A study of the extracellular nitrogen of several molds' gave 
the results in the accompanying table. 



TABLE ill 
Amounf of Nitrogen Assitnilafed Which Appeared in fhe Medium After Seven Days Growth 



Fungus 


Nitrogen 
source 


Extracellular nitrogen 

(as % initially added 

nitrogen) 




NH4 + 

NO.r 
NH4+ 
Nor 

NH4+ 

NOr 
NH4+ 


7.5 
3.5 




20.0 
7.5 




27.0 
36.0 




23.5 







The extracellular nitrogen was related to the nitrogen supplied 
in two cases: 



TABLE IV 
Formation of Extracellular Nitrogen in Relation to Initially Added Nitrogen Which 

Disappeared 





Nitrogen 
Source 


Amount of nitrogen supplied (mg. /flask) 


Fungus 


6.6 13.2 6.6 + 6.6 




Extracellular N as % N assimilated 


Scopulariopsis brevicaulis 

Penicillium griseofulvum 


NH4- 
NOr 
NO3- 


25.35 ± 5.58 25.80 ± 2.86 

20.03 ± 4.47 1 6.62 ± 2.08 1 6.50 ± 1 .98 

1 3.00 ± 3.35 1 2.50 ± 0.70 1 2.34 ± 2.73 



M. Richards and R. H. Haskins, Can. J. Microbiol. 3 543 (1957). 
A. G. Morton and D. Broadbent, 7. Gen. Microbiol. 12 248 (1955). 



303 



Amino Acids and Related Compounds 



In this earlier study most of the extracellular nitrogen appeared 
to be peptide in nature, yielding some 14 amino acids on hy- 
drolysis. In the one case tested one of the fungi was unable to 
use the extracellular nitrogen formed, but assimilated the con- 
stituent amino acids when these were liberated by acid hydroly- 
sis. 

A quantitative report has been made on the free amino acids 
present in an alcohol extract of Miicor miLcedo.^ They were as 
follows : 

TABLE V 
Amino Acids Present in 75% Alcohol Extracfs of Mucor mucedo (as % Tofai Nitrogen) 



Amino acid 




Hydrolysate of 
insoluble residue 



Alanine 

/3-Alanine 

Arginine 

Asparagine 

Aspartic acid 

-)-Aminobutyric acid 

Citrulline 

Cystine 

Giutamine 

Glutamic acid 

Glycine 

Histidine 

iso-Leucine 

Leucine 

Lysine 

Methionine 

Proline 

Phenylalanine 

Serine 

Threonine 

Tyrosine 

Valine 



6.7 
12.2 
16.4 

2.2 

35.2 
3.8 
1.6 
0.7 
5.2 
3.8 
1.1 
4.2 
1.0 
3.3 
2.2 
2.1 
4.4 



These values were compared with those of other plants over a 
taxonomic range. 

A report of the free amino acids produced by Penicillium 
roquefortii indicated the following to be most prominent:^ 

^K. Mansford and R. Raper, Nature 174 314 (1954). 
^ J. Kolousek and S. Michalik, Sbornik Ceskoslov. Akad. Zemedel 
Ved. 27A 281 (1954). (Chem. Abstr. 50 4295c) 



Pfizer Handbook of Microbial Metabolites 



304 



Aspartic Acid 
Glutamic Acid 
Serine 
Threonine 
a-Alanine 



Valine 
Methionine 
Leucine 
Isoleucine 



The free and combined amino acids of the uredospores of 
ten wheat rust strains have been determined quantitatively.^" 

The intracellular amino acids of microorganisms have been 
studied. Gale demonstrated the presence of such a pool in 
Streptococcus faecalis}^ Gale and Taylor extended the investi- 
gation to a variety of bacteria and yeasts with particular atten- 
tion to lysine and glutamic acid.^- Fuerst studied several 
fungi." The free intracellular amino acids of certain strains 
of Neurospora crassa have been explored." The relative quan- 
tities of amino acids present varied widely among the various 
mutants. In all some 35 ninhydrin-positive substances were en- 
countered among the 28 different strains studied. The free 
amino acids of Staphylococcus aureus have been determined, 
and the ability of bacteria to concentrate amino acids strikingly 
demonstrated by comparison of the concentrations of internal 
and external acids. ^^ 



TABLE VI 
Free Amino Acids in Exponenfially Growing Staphylococcus aureus Cells Growing in Synfhefic 

Medium 



Amino Acid 


Quantity (jumoie/g.) 
in internal pool 


Ratio of internal to 
external concentration 




39.6 

38 

16.8 
8.3 
2.6 
6.7 


25.4 




22.6 




23.2 




13.3 




4.5 




8.3 







1" M. E. McKillican, Can. J. Chem. 38 244 (1960). 

"E. F. Gale, /. Gen. Microbiol, 1 53 (1947). 

12 E. F. Gale and E. S. Taylor, ibid. I 77 (1947); E. S. Taylor, ibid. 
1 86 (1947). 

" R. Fuerst and J. Awapara, Texas Repts. Biol, and Med. 10 424 
(1952). 

^^ Robert Fuerst and Robert P. Wagner, Arch. Biochem. and Bio- 
phys. 70 311 (1957). 

15 R. Hancock, Biochim. et Biophys. Acta 28 402 (1958). 



305 



Amino Acids and Related Compounds 



TABLE VI — Continued 
Free Amino Acids in Exponentially Growing Sfaphylococcus aureus Cells Crowing in S/nfhef/c 

Medium 



Amino Acid 



Alanine 

Cystine and cysteine 

Serine , 

Glycine 

Tyrosine 

Lysine < 

Arginine ■ 

Histidine , 

Phenylalanine 

Threonine , 

Tryptophan , 



Quantity (^umole/g.) 


Ratio of internal to 


in internal pool 


external concentration 


8.1 


5.4 


5.5 




3.4 


5.4 


2.8 


2.3 


2.4 


3.1 


2.2 


4.6 


2.2 


3.0 


1.7 


2.2 


1.3 


1.8 


1.0 


1.6 


0.3 





All the amino acids found in the internal protein of the cell 
were present in the internal pool of free amino acids. 

A new amino acid, S-methyl-L-cysteine, has been isolated 
from Neurospora crassa.'^*' An isomer of ^-methyllanthionine 
has been isolated from yeast." Urocanic acid has been detected 
in Micrococcus lysodeikticus}^ 



CHa— S— CH2— CH— COOH 

NHo 
S-Methylcysteine 



HOOC— CH— CH2— S— CH— CH— COOH 

NHo CH3 NH2 

/3-Methyllanthionine 



HOOC— CH—CH— COOH 

NH2 NH2 
a,/3-Diaminosuccinic Acid 

New, partially characterized a-amino acids have been isolated 
from boletus and lactarius species. ^'''' ^"^ -^ a,/3-Diaminosuccinic 
acid has been isolated from production filtrates of the antibiotic 

^•^ James B. Ragland and James L. Liverman, Arch. Biochem. and 
Biophys. 65 574 (1956). 

"PhyUis F. Downey and Simon Black, /. Biol. Chem. 228 171 
(1957). 

'^^ Jana Gregoire and Jean Gregoire, Compt. rend. 245 2553 (1957). 

^^ A. I. Virtanen and O. Ayrapaa, Suomen Kern. SIB 190 (1958). 

20 Atsushi Komamine and Artturi I. Virtanen, Acta Chem. Scand. 
13 2141 (1959). 

21 J. Casimir and Artturi I. Virtanen, ibid. 13 2139 (1959). 



Pfizer Handbook of Microbial Metabolites 306 

oxytetracycline (Streptomyces rimosiis).-^ The structures of 
certain other unusual amino acids are listed in this section. 

Production of glutamic acid by streptomycetes on synthetic 
medium containing glycine has been investigated.-' Yields of 
extracellular glutamic acid were 0.25-1.75 g. per liter. It was 
the only amino acid or nitrogenous material present after four 
and seven days, but after ten days some alanine, phenylalanine, 
aspartic acid and glycine appeared. Strains examined were: 
Streptomyces anmilatus, S. aureofaciens, S. fradiae, S. olivaceus 
and S. rimosus. 

The high proportions and amounts of L-glutamic acid syn- 
thesized by microorganisms have led to the development of an 
economical process for its commercial production. Certain mi- 
crococcus and bacillus species produce more than a 20% yield 
(molar basis) from the glucose supplied.-^ A similar yield of 
valine has been reported.-"' 

L-Lysine is also produced commercially by a direct process 
(micrococcus, bacillus)-'* and by a two-stage process (Escher- 
ichia coli, Aerobacter aerogenes),-' 2,6-diaminopimelic acid be- 
ing the intermediate in the latter case. 

Tryptophan production by E. coli and by Salmonella typhi has 
been reported as small unless indole is added.-'' The indole 
apparently competitively inhibited tryptophanase. Many mi- 
croorganisms are able to synthesize tryptophan from indole and 
serine. 

A survey of 20 genera, 72 species and 334 strains of aerobic 
bacteria for amino acid accumulation revealed no marked tax- 
onomic difference except that facultative aerobes such as escher- 
ichia, aerobacter and bacillus species were superior to obligatory 
aerobes such, as pseudomonas. Production and accumulation 
were more dependent on strain and conditions.-" 

The biosynthesis and metabolic interrelationships of amino 
acids can be considered here only in briefest summary because 
of the breadth and complexity of the subject. More thorough re- 

22 F. A. Hochstein, /. Org. Chem. 24 679 (1959). 

23 D. Perlman and E. O'Brien, /. Bad. 75 611 (1958). 

-^ Toshinobu Asai, Ko Aida and Kunio Oishi, Bull. Agr. Chem. Soc. 
(Japan) 21 134 (1957). 

25 Zenjiro Sugisaki, Nippon Nogei-kagakii Kaishi -54 153 (1960). 

2" Shukuo Kinoshita, Kiyoshi Nakayama and Sohei Kitada, J. Gen. 
Appl. Microbiol. 4 128 (1958). 

27 Lester E. Caslda, Jr., U. S. Patent 2,711,396 (1956). 

2sp. Fildes, J. Gen. Microbiol. 15 636 (1956). 

2" Hiroshi lizuka and Kazuo Komagata, Nippon Nogei-kagashu 
Kaishi 34 27 (1960). 



307 



Amino Acids and Related Compounds 



views are available*"' ^^' ■*- and references to some of the vast 
literature on this subject can be found there. 

The occurrence studies cited demonstrate the importance of 
glutamic acid. It is a constituent of folic acid and related sub- 
stances, and of glutathione, and various antibiotics. It occurs 
in the cell wall of bacteria and, as a polypeptide, is the sole 
capsular substance of certain bacilli. Its wide distribution re- 
flects its cross-roads position in nitrogen metabolism. 

Synthesis of glutamic acid by most aerobic microorganisms 
involves ainination of a-ketoglutaric acid (a reversible reac- 
tion), thus tying it in with the citric acid cycle. It is a pre- 
cursor of ornithine, proline and in some cases lysine. 

O 

I amination 
HOOC— CHo— CH,— C— COOH ;:^ ^ HOOC— CHo— CH2— CH— COOH 

a-Ketoglutaric Acid I 

NH2 

Glutamic Acid 

In E. coli, at least, the route to ornithine involves N-acetylated 
intermediates. The intermediates shown accumulate in appro- 
HOOC— CHo— CH2— CH— COOH 



NH2 

Glutamic Acid 






reduction 






OHC— CH2— CH2— CH— COOH _-^ 


CH2— CH2 


NH2 

Glutamic Acid Semialdehyde 

. . I 
amination 




CH CH— COOH 

N 

A^-Pyrroline-5- 
carboxylic Acid 


H2N— CH2— CH2— CHo— CH- 


-COOH 


reduction 
CH2— CH2 


NHo 
Ornithine 








CH2 CH— COOH 

\ / 
N 
H 



Proline 

30 Bernard D. Davis, Advances in Enzymol. 16 247-312 (1955). 

^'^ Alton Melster, "Biochemistry of the Amino Acids," Academic 
Press, New York, 1957, pp. 256-394. 

■^-Joseph S. Fruton and Sofia Simmonds, "General Biochemistry," 
John Wiley and Sons, Inc., New York, 1958, pp. 771-896. 



Pfizer Handbook of Microbial Metabolites 308 

priate auxotrophs and can be isolated. This scheme has been 
found in a variety of molds, yeasts and bacteria. 

Ornithine reacts with carbamyl phosphate to form citrulline, 
an intermediate in the biosynthesis of arginine: 

H2N— CH2— CH2— CH2— CH— COOH 

I 

NH2 
Ornithine 

O 

I H2N— C— O— PO3H2 

o 

II 

H2N— C— NH— CH2— CH2— CH2— CH— COOH 

I 
NH2 

Citrulline 

HOOC— CH— CH2— COOH 



NH2 

e 

00c— CH—CH2— COOH 

I 
©NH 

H2N— C— NH— CH2— CH2— CH2— CH— COOH 

I 
NH2 

Argininosuccinate 

^ > HOOC— CH=CH— COOH 

NH ^ 

II 
H2N— C— NH— CH2— CH2— CH2— CH— COOH 

I 

NH2 
Arginine 

Arginine can complete the "urea cycle" by losing urea to form 
ornithine. Enzymes for all these steps have been found in var- 
ious microorganisms. 

Glutamic acid acts as an ammonia carrier by formation of its 
half amide, glutamine, and in this way contributes nitrogen to 
the biosynthesis of purines and amino sugars. 

Aspartic acid also occupies a central position in nitrogen 
metabolism. In microorganisms it can be synthesized either by 
amination of oxaloacetic acid or by the addition of ammonia 



309 Amino Acids and Related Compounds 

to fumaric acid, the former process probably being more preva- 
lent. 

O NH2 



HOOC- 


-c- 


-CHj— COOH 


amina 


Hon 


HOOC- 


-CH- 


-CHo- 


-COOH 


Oxc 
HOOC- 


iloacetic Acid 
-CH=CH— COOH 


NH3 




Aspartic Acid 
NH.. 

HOOC— CH— CH>- 


COOH 


F 


jmaric Acid 






A 


spartic Acid 





Either equation ties aspartic acid in with the citric acid cycle. 
Like glutamic acid, aspartic acid acts as an ammonia carrier 
through its half amide, asparagine. One role of aspartic acid 
was seen above in the biosynthesis of arginine. Aspartic acid 
has been proved a precursor of pyrimidines in certain microor- 
ganisms. It is also a precursor of threonine and of both a- and 
/3-alanines. Separate enzymes control the selective decarboxyla- 
tions to form the alanines. 





-CO2 




HOOC— CH2— CH— COOH - 

1 


> 


CH3— CH— COOH 

1 


NH2 X,^^ 




NH2 


Aspartic Acid 


X-COz 


a-Alanine 

HOOC— CH2— CH2— NH2 
j3-Alanine 



a-Alanine (either isomer) can be synthesized, too, from py- 
ruvic acid by a wide variety of biological systems. Some mi- 
croorganisms effect this amination directly from ammonia, but 
the transamination from glutamate is probably more prevalent. 
Alanine, therefore, is also closely connected with carbohydrate 
and fat metabolism, and it is used as an energy source by many 
microbes. Through pyruvate it also may be considered a pre- 
cursor of glycine, serine, cysteine and of valine, leucine and 

O NH2 

CH3— C— COOH + HOOC— CH—CH2—CH2— COOH ^ CH3— CH— COOH + 

NHo 
Pyruvic Acid Glutamic Acid a-Alanine 

O 

I 

HOOC— C—CH2—CH2— COOH 
a-Ketoglutaric Acid 



Pfizer Handbook of Microbial Metabolites 310 

isoleucine. D-Alanine occurs in bacterial cell walls and spores 
and frequently in antibiotics. Some bacteria even require an 
exogenous source of D-alanine, particularly on a medium devoid 
of pyridoxine, since pyridoxal phosphate is a coenzyme for the 
racemase. ^-Alanine is a component of coenzyme A. A re- 
lated substance, ^-nitropropionic acid has been isolated from an 
aspergillus species. 

Glycine and serine are reversibly interconvertible in most or- 
ganisms, tetrahydrofolic acid transferring the hydroxymethyl 
group. Glycine also is formed by amination of glyoxylate in 
some microorganisms. 

pyridoxal phosphate 
THFA CH2OH 
CH2— COOH ( : =± HOCH2— CH— COOH 

I I 

NH2 NH2 

Glycine Serine 

glutamate 
or 

ammonia 
OHC— COOH — ==^ NHo— CH2— COOH 
Glyoxylic Acid Glycine 

In E. coll serine is probably to be regarded as the precursor of 
glycine. The origin of serine is still obscure. There is a possi- 
bility that it may arise from phosphoglyceric acid from the gly- 
colysis scheme: 

COOH COOH COOH COOH 
CH— OH ;=iC=0 ;=±CH— NH2 ^ CH— NH2 

CH2OPO3H. CH2OPO3H2 CH2OPO3H2 CH2OH 

3-Phospho- 3-Phospho- Phospho- Serine 

glyceric hydroxy- serine 

Acid pyruvic Acid 

Glycine is a precursor of the porphyrins, purines, glutathione 
and sarcosine. 

Serine contributes the carbon skeleton of cysteine in most or- 
ganisms. Most microorganisms can use sulfate but not methio- 
nine as a sulfur source, while mammals require methionine for 
this purpose but cannot use sulfate. The conversion route of 
methionine to cysteine has been worked out for higher animals, 
but is not entirely understood in microorganisms. 

Thiosulfate is used by some molds, and cysteine-S-sulfonate 
has been found to be an intermediate. Hydrogen sulfide has 
been reported as a precursor in yeast. Threonine has been iso- 
lated as an intermediate to cysteine in a neurospora auxotroph. 



311 



Amino Acids and Related Compounds 



Cysteine is a component of glutathione and of penicillin. 
Methionine is important in transmethylation reactions. The 
entire topic of one-carbon metabolism cannot be reviewed here. 
The transfer of methyl groups from methionine to oxygen and 
nitrogen atoms, and probably to carbon atoms in biosynthetic 
sequences requires ATP, and the active complex has been iden- 
tified as S-adenosylmethionine. 

NH2 



^'^^M 



© "^\ Jl J 

OOC— CH— CH2— CH2— S— CH2 ^N-^-N^ 
NH2 CH3 




OH OH S-Adenosylmethionine 

Several labile methyl group compounds (choline, betaine, ser- 
ine) probably can contribute the methyl group of methionine 
by way of the proper coenzymes (Bj^, THFA). Some neurospora 
mutants have been found which seem to synthesize methionine 
from cysteine and, ultimately, from aspartic acid. The follow- 
ing scheme has been suggested: 

ATP TPN 
HOOC— CH,— CH— COOH > H2O3P— OOC— CH.— CH— COOH > 



I 

NH2 
Aspartic Acid 

OHC— CH,— CH— COOH 

NH.j 
Aspartic ^-Semialdehyde 



TPN 



NHo 
/i-Aspartyl Phosphate 

HOCH2— CHo— CH— COOH, 

NH.. 
Homoserine 



/ 



HS— CH>— CH,— CH— COOH CH,— CH,— CH— COOH 

NH, S NH2 

Homocysteine CH,— CH— COOH 

NH, 
Cystathionine 

CH3— S— CH2— CH2— CH— COOH 



CH3— CH— CH— COOH 

OH NH2 
Threonine 



CH2SH 
CH— NH2 

COOH 

Cysteine 



NH2 



Methionine 



Pfizer Handbook of Microbial Metabolites 312 

Homoserine also is a precursor of threonine in neurospora 
mutants, with ATP and pyridoxal phosphate required. Threo- 
nine is synthesized by most microorganisms although it is an 
essential in mammalian diets. 

The fact that lysine-requiring neurospora mutants use a-ami- 
noadipic acid makes probable a biosynthetic scheme in which 
the terminal carboxyl group is reduced and aminated as in the 
biosynthesis of ornithine from glutamic acid. Some molds even 
are able to use a-ketoadipic acid, which strengthens the argu- 
ment. Labeling studies indicate formation of the a-ketoadipic 
acid by condensation of acetate with either a-ketoglutarate or 
with the "active succinate" from the citric acid cycle, the acetate 
carboxyl furnishing the carboxyl group of lysine. Proposed 
lysine biosynthesis in molds: 



HOOC— CHo— CH2— CO— CoA(COOH) HOOC— CH2— CH2— CH2— C— COOH 
Active Succinate — > a-Ketoadipic Acid 

CH3-CO-C0A aminationj 

Acetate 

HOOC— CH2—CH2—CH2—CH— COOH 

I 

NH2 
OCH— CH2— CH2— CH2— CH— COOH < a-Aminoadipic Acid 



NH2 



H2N— CH2— CH2— CH2— CH2— CH— COOH 

NH2 



a-Aminoadipic Acid 
«-Semialdehyde 

amination 



Lysine 

a-Aminoadipic acid is produced by Penicillium chrysogenum as 
a component of a tripeptide isolated from the mycelium. It also 
occurs as a moiety of the antibiotic synnematin-B (cephalo- 
sporin N) produced by the mold Cephalosporium salmosynne- 
matum, and it has been isolated from Aspergillus oryzae. 

a,€-Diaminopimelic acid is a precursor of lysine in E. coli and 
in many other bacteria. L,L-Diaminopimelic acid is formed in 
E. coli by condensation of pyruvic acid with aspartic acid. 
Later a specific racemase converts it to the meso-form. A com- 
plete mechanism for lysine biosynthesis in bacteria has been 
proposed : 



313 



Amino Acids and Related Compounds 



COOH 
CH— NH, 


HOOC 


— CH 

Succ 


2— CH,— COOH 
nic Acid 


^COOH 
CH— NH 


-CO— CH2- 


-CH2- 


-COOH 


CHo 
COOH 






ATP 


> 


CH, 

c=o 








Aspartic CH3 
Acid 1 

C= 











CH2 

c=o 








COOH 
Pyruvic Acid 








_COOH 


1 




- 


COOH 










COOH 








CH— NH 


— CO- 


-CH:- 


-CH,- 


COOH 


CH— NH- 


-CO— CH2- 


-CH2- 


-COOH 


CHo 










CH2 








CH2 








- 


>CH2 








CH2 










CH2 








c=o 










CH— NH2 

1 








COOH 










COOH 









CH2— NH2 




\ 






COOH 




COOH 


—CO 


2 






/-u < 




+ 




^n2 < 






1 


CH— NH2 




CH, 


CH2 


1 




1 


1 


CH, 




CH, 


CH2 


1 




1 


1 


CH, 


COOH 


CH— NH, 


1 




Succinic 


1 


CH, 




Acid 


COOH 


1 






Lysine 


CH— NH, 
COOH 








Diaminopimeiic 






Acid 







Intermediate III has been isolated and identified, and there is 
some evidence for the existence of II. Rather similar interme- 



Pfizer Handbook of Microbial Metabolites 314 

diates have been suggested as precursors of 2,6-dipicolinic acid, 
which is formed in some bacterial spores. Free diaminopimelic 

O 

II 

c 




CHo CHo 

HOOC— I I — > /^N- 

CH C HOOC COOH 

\ ■i^\ Dipicolinic Acid 

NH2 O COOH 

acid has been isolated from vegetative cells of such spore-form- 
ers. It has never been found in yeasts and molds. 

a,£-Diaminopimelic acid replaces lysine in the repeating pen- 
tapeptide unit of the bacterial cell wall in Corynehacterium 
diphtheriae , E. coli and certain other bacteria (especially gram 
negatives). Some E. coli strains accumulate considerable quan- 
tities of diaminopimelic acid, and this faculty has been ex- 
ploited in a two-step commercial production process. 

Many microorganisms metabolize lysine to pipecolic acid, a 
component of several antibiotics. 



CH2 



CH2 CH2 



-"' o. 



CHo CH— COOH ^N' 

\ / H COOH 

_ NH2 NH2 Pipecolic Acid 
Lysine 

The amino acids discussed to date are closely integrated with 
carbohydrate and fat metabolism. Those remaining to be con- 
sidered are more remotely derived. 

Valine, isoleucine and leucine are essential to the mammalian 
diet and are required also by many microorganisms. This 
seems to indicate enzymatic difficulties in the biosynthesis of 
these branched-chain amino acids. 

Much evidence has accumulated concerning the biosynthesis 
of valine and isoleucine, and the following pathway is indi- 
cated (for valine): 



315 



Amino Acids and Related Compounds 



O OH 



CH3— C— COOH > CH3— C— C— COOH 

(3) ® ® CH3CHO® ® I®® 

Pyruvic Acid ® ® CH3 

"active acef- ® 

aldehyde" a-Acetolactic Acid 

thiamine 
O' pyrophosphate 

II 
CH3— C— COOH 



rearrangement 



O OH OH 

II H2O I I TPNH 
CH3— CH— C— COOH <- CH3— C— CH— COOH < 



CH3 
a-Ketoisovaleric 
Acid 
transamination 



reduction 



CH3 
a,/3-Dihydroxy- 
isovaleric Acid 



OHO 

CH3— C— C— COOH 

® ® I @ ® 

CH3 
® 
a-Keto-/3-hydroxy- 
isovaleric Acid 



CH3 



CH— CH— COOH 



CH3 



NH2 
Valine 



The intermediates, a-acetolactic acid and a,^-dihydroxyiso- 
valeric acid, have been isolated from a variety of microorgan- 
isms and are well characterized. a-Keto-^-hydroxyisovaleric 
acid has not been reported yet, although when it is mixed with 
enzyme preparations from molds and yeasts together with 
TPNH, it is reduced to a,^-dihydroxyisovaleric acid. a-Ketoiso- 
valeric acid is aminated by numerous microorganisms. 

The scheme for isoleucine is believed to be analogous, but 
with a-ketobutyric acid replacing pyruvic acid as the initial sub- 
stance. This four-carbon acid is, in turn, derived from homo- 
serine or threonine, and ultimately from aspartic acid. Some 
of the steps of the valine and isoleucine syntheses are known to 
share common enzymes. 

Leucine biosynthesis is apparently the same as that of valine 
up to the final amination step. Leucine, however, requires 3 
moles of pyruvate for its 6-carbon atom chain rather than the 



Pfizer Handbook of Microbial Metabolites 



316 



2 required by valine. The remaining steps of the proposed 
leucine biosynthesis in microorganisms are; 



CH3 O 

\ II -CO, 

CH— C=0 + CH3— C— COOH -* 

/ I Pyruvic Acid [O] 

CH3 COOH 

a-Ketoisovaleric 
Acid 



"CHa 



CH3 



CH3 CH3 

\ 

CH — CH2— CH— COOH amination 

/ I < 

CHs 



OH 

I 
CH— C— CH2— COOH 

' I 

COOH 



CH— CH2— C— COOH*- 



CH3 



NH2 
Leucine 



a-Ketoisocaproic Acid 

This partial scheme is based on labeled media experiments in 
yeasts, molds and bacteria. 

The biogenetic scheme of the aromatic amino acids phenyl- 
alanine and tyrosine was briefly outlined in the introduction to 
the section on simpler ahcyclic compounds. The final stages 
of this route are shown here, beginning with shikimic acid: 




COOH Pyruvic Acid 

^> V 



HO 



CH2— CO— COOH 



H — COOH 
Prephenic Acid 



Shikimic Acid 
5-Phosphate 



HO 



_ p-Hydroxyphenyllactic Acid 




-CH2— CH— COOH 



CH2— CO— COOH 

Phenylpyruvic Acid 
kNH3 



f2H 

HO— f V-CH2— CO— COOH ( \-CH2— CH— COOH 

\=/ \=/ I 

p-Hydroxyphenylpyruvic Acid NH2 

l^NHs Phenylalanine 



HO 



i 



CH2— CH— COOH 



NH2 



Tyrosine 



3^7 



Amino Acids and Related Compounds 



The benzene ring of tryptophan also arises from the shikimic 
acid route. The intermediates are unknown between shi- 
kimic acid and the first aromatic member of the sequence, 
anthranilic acid: 

NHo 




— COOH 



:OOH 
Anthranilic Acid 



Shikimic Acid 



The remainder of the sequence in its present state of devel- 
opment is as follows: 




COOH 5-Phosphoribosyl- 
1 -pyrophosphate 



COOH 



NH2 
Anthranilic Acid 




CH— CH— CH— CH— CH2— O— PO3H2 

NH OH OH 

N-(2-Carboxy phenyl)-! -aminoribose-5-phosphate 



Co 



^CH— CH— CH2 

I I I 
OH OH O— PO2H2 




Triose- ^^ 
phosphate 



lndolyl-3-glycerol 
Phosphate 



L-Se 



Amadori rearrangement 
COOH 



NH— CH2— CO— CH— CH— CH2— OPOaHs 



OH OH 

1 -Deoxy-1 -(o-carboxyanilino)-ribulose 



,CH.— CH— COOH 

I 
NH2 



Indole Tryptophan 

There appears to be some question as to whether the Amadori 
rearrangement product is a bona fide member of this sequence. 
It has been isolated from Aerobacter aerogenes and character- 
ized as derivatives, and it substitutes for anthranilic acid in bac- 
terial mutants requiring the latter. 

The anthranilic acid carboxyl group is known to be lost as 
carbon dioxide during the formation of the pyrrole ring, and 
the first two carbon atoms of ribose are known to form the 2 
and 3 positions of the indole ring. Glucose also can furnish 
these two carbon atoms. In this connection it should be men- 



Pfizer Handbook of Microbial Metabolites 



318 



tioned that N-fructosylanthranilic acid has been isolated from 
a yeast. Probably indole never exists in the free state to any 
appreciable extent during the tryptophan synthesis, but is en- 
zyme-bound. 

Ribose contributes also to the biosynthesis of histidine. Here 
purines are catalytic, furnishing a carbon atom and a nitrogen 
atom from the pyrimidine ring to form positions 2 and 3 of the 
histidine ring. The purine is then regenerated by reaction with 
a Ci substance. Adenine is the most efficient purine for this 
purpose. The following scheme has been worked out, largely 
on the basis of auxotroph work: (P = phosphate, R = ribose). 



HO 

CH, 

I 
CH2— CH— CH— CH 



NH2 

I Ribose-5- vik ^ 

^j:?:\^N. phosphate | ^^ 

I iT )> ^ CH2— CH— CH— CH I 



NH2 



ATP 



P— O OH OH OH 



R— P 
Adenosine 
Monophosphate 



Ci substance, 
reamination 



R— P 
Glutamine 



HC- 



CH>— CH— CH— C- 



P— O OH OH 

4-(D-ery tfiro- 1 ',2'-dihydroxy- 
3'-phosphopropyl) imidazole 
(Imidazoleglycerol Phosphate) 




R— P 

5-Amino-l-D- 

(5'-phospho- 

ribosyl)-4- 

imidazolecarboxamide 



Some chemicals which inhibit purine synthesis also cause ac- 
cumulation of such intermediates. To continue with the bio- 
synthesis of histidine: 



319 Amino Acids and Related Compounds 

HC C CH— CH-CH,— O— P H,0 HC=C— CH.— C— CHj— O— P 



N NH OH OH -^-^ N NH O 

c c 

H H 



Imidazole glycerol Imidazoleacetol 

Phosphate Phosphate 



glutamate 

— » a-ketoglutarate 



H3PO, + 



HC=^C— CHo— CH— CH,— OH H,0 HC=— C— CH,— CH— CH2— O— P 

N NH NH2 ^ '^— N NH NH2 

\/ \/ 

C C 

H H 

L-Histidinol L-Histidinol 

2DPN 



2DPNH 



HoO Phosphate 

© 
2H 



HC=C— CH,— CH— COOH 

N NH NH> 

\/ 
C 
H 

Histidine 
It is interesting that the final stages of this synthesis differ 
from those in the tryptophan sequence when some of the inter- 
mediates are so closely related. Perhaps in some species a 
lesser difference will be found. 

Histidine is converted to ergothioneine in microorganisms by 
methylation to form hercynine, followed by direct introduction 
of the thiol group. 

663 Glycine, C.H,,O.N, colorless crystals, m.p. -'280-290° (dec.) 

(rapid heating). 

H,N— CH,— COOH 
Widely distributed. 

664 Sarcosine, C3H7O0N, colorless crystals, m.p. 212° (dec). 

CH,NHCH,COOH 

Cladonia sylvatica 

Also a component of the actinomycin antibiotics. 
P. Linko, M. Alfthan, J. K. Miettinen and Artturi I. Virtanen, 
Acta Chem. Scarid. 7 1310 ri953). 



Pfizer Handbook of Microbial Metabolites 320 

665 L-Alanine, C3H7O2N, colorless crystals, m.p. 297° (dec), [cnW^ 

+8.5° (9.3% solution of the hydrochloride in water). 

CH3— CH— COOH 
NH2 
Widely distributed. 

666 /^-Alanine, C3H7O2N, colorless crystals, m.p. 207° (dec.) (pre- 

heated bath). 

H2N— CH2— CH2— COOH 
Widely distributed. 

667 L-Serine, C3H7O3N, colorless crystals, m.p. 228° (dec.) (sub- 

limes 150° at 10-* mm. Hg), [aW +14.45° (0.5 g. per 
5.6 g. of 1 N hydrochloric acid). 

HO— CH2CH— COOH 

I 
NH2 

Widely distributed. 

668 L-Aspartic Acid, C4H7O4N, colorless crystals, m.p. 270° (sealed 

capillary, preheated bath) (dec), [a]D-* +24.6° (c 2 in 6 N 
hydrochloric acid). 

HOOC— CH2— CH— COOH 
NH2 
Widely distributed. 

669 L-Asparagine, C4H8O3N2, colorless crystals ( Monohydrate ) , m.p. 

234°, (dec) (preheated bath), [ajn'" -5.5° (c 1.3 in wa- 
ter). 

O 
H2N— C— CH2— CH— COOH 
NH2 
Widely distributed. 



321 Amino Acids and Related Compounds 

670 d-Diaminosuccinic Acid, C4HSO4N0, colorless crystals, m.p. 

(dec), 240-290°, [aW +28° (c 2.0 in 5% sodium hy- 
droxide solution). 

HOOC— CH— CH— COOH 

I I 
NH2 NH2 

Streptomyces rimosus 

This amino acid sometimes crystallizes from oxytetra- 
cycline broth concentrates. The yield is about 250-500 
mg. per liter. 

F. A. Hochstein, /. Org. Chem. 24 679 (1959). 

671 O-Carbamyl-D-serine, C4H8O4N2, colorless needles, m.p. 226- 

234° (dec), [ah -19.6° (c 2 in N hydrochloric acid). 

O 

il 
H2N— C— O— CHo— CH— COOH 

I 
NH2 

Streptomyces polychromogenes 

D-Serine or derivatives is also present in polymyxin, 
echinomycin, cycloserine and amicetin. 

G. Hagemann, L. Penasse and J. Teillon, Biochim. et Bio- 
phys. Acta 17 240 (1955). 

672 Allantoic Acid, C4HSO4N4, colorless needles, m.p. 165° (dec). 

H2NCONH 

\ 

CH— COOH 

/ 
H2NCONH 

Coprinus miraceus, Collybia dryophila 

R. Fosse and A. Brunei, Compt. rend. 197 288 (1933). 

673 y-Aminobutyric Acid, C4H9O2N, colorless crystals, m.p. 202° 

(dec) rapid heating. 

H2N— CH2— CH2— CH2— COOH 
Widely distributed. 



Pfizer Handbook of Microbial Metabolites 322 

674 L-(+)-a-Aminobutyric Acid, C4H9O0N, colorless crystals, m.p. 

270-280° (dec), [a]ir" +8.0° (c 1.0 in water). 

CH3— CHo— CH— COOH 

I 
NH2 

Escherichia coli, Corynebacterium diphtheriae 

A. Poison, Nature 161 351 (1948). 

Elizabeth Work, Biochim. et Biophys. Acta 3 400 (1949). 

675 L-Threonine, C4H9O3N, colorless crystals, m.p. 255-257° (dec), 

[a]D-'" -28.3° (c 1.1 in water). 

CH3— CH— CH— COOH 

I I 
OH NHo 

Widely distributed. 

676 S-Methyl-L-cysteine, C4H,,03NS, colorless crystals, m.p. --164° 

(dec), [alo'' +125° (c 2.5 in water). 

CH3— S— CH>— CH— COOH 

NHo 

Neurospora crassa 

James B. Ragland and James L. Livermore, Arch. Biochem. 
and Biophys. 65 574 (1956). (Isolation from neurospora) 

Clayton J. Morris and John P. Thompson, /. Am. Chem. 
Soc. 78 1605 (1956). (Physical properties) 

677 4-Imidazolyacetic Acid, C-.H^OoN^, colorless needles (Hydrate), 

m.p. 222° (dec). 

N C— CH2— COOH 

II II 

HC CH 

H 

Polyporus sulfureus 

P. H. List, Planta Med. 6 424 (1958). 

678 Azaserine (Diazoacetyl-L-serine), C-,H704N;^, light yellow-green 

crystals, dec 146-162°, [a],,"' '' -0.5° (c 8.46 in water at 
pH 5.18). 

N.— CH— CO— O— CH,— CH— COOH 

NH, 
An unclassified streptomycete 



323 Amino Acids and Related Compounds 

James A. Moore, John R. Dice, Ernest D. Nicolaides, Roger 
D. Westland and Eugene L. Wittle, /. Am. Chern. Soc. 7(i 2884 
(1954). (Synthesis) 

C. Chester Stock, H. Christine Reilly, Sonja M. Buckley, 
Donald A. Clarke and C. P. Rhoads, Nature 173 71 (1954). 

John Ehrlich, Lucia E. Anderson, George L. Coffey, Arthur 
B. Hillegas, Mildred P. Knudsen, Harold J. Koepsell, Dorothy 
L. Kohberger and Julian E. Oyaas, ibid. 173 72 (1954). 

Quentin R. Bartz, Carole C. Elder, Roger P. Frohardt, Salva- 
tore A. Fusari, Theodore H. Haskell, Doris W. Johannessen and 
Albert Ryder, ibid. 173 72 (1954). (Isolation) 

679 L-Proline, C.^HciOoN, colorless crystals, m.p. 220-222° (dec.) 

(rapid heating), [x],,-"' —80° (c 1.0 in water). 

CH. — CH2 

CH2 CH— COOH 

H 

Widely distributed. 

680 L-GIutamic Acid, C-,H.,04N, colorless crystals, m.p. 247° (dec), 

[a]n--* +31.4° (c 1 in 6 N hydrochloric acid). 

HOOC— CH2— CHo— CH— COOH 

NH2 

Micrococcus varians 

A 17% molar yield (from glucose) was reported. 
Toshinobu Asai, Ko Aida, Kunio Oishi, Bull. Agr. Chem. Soc. 
(Japan) 21 134 (1957). 

681 L-Glutamine, C.-jHjoO^N^, colorless crystals, m.p. 185° (dec), 

[a]ir' +5.9° (c 4.0 in water). 

O 

H ,N— C— CH2— CH2— CH— COOH 

i 
NH2 

Widely distributed. 

682 L-Valine, C-,Hi,O.N, colorless crystals, m.p. 315° (dec.) (closed 
capillary). Sublimes, [a]ir'' +14°(c0.9in water). 

CH3 

\ 

CH— CH— COOH 

/ I 

CH3 NH2 



Pfizer Handbook of Microbial Metabolites 324 

Widely distributed. 

683 Betaine, C5H11O2N, white prisms or leaflets, m.p. 293° (dec.)- 

© e 

(CHalsN— CH2COO 

Aspergillus oryzae, Patella vulgata, Claviceps purpurea 
(Fries) Tul. and other fungi 

Jacqueline Etienne-Petitfils, Bull. soc. chim. biol. 38 1315 

(1956). 

684 L-Methionine, CgHnOgNS, colorless crystals, m.p. '-'280° (dec.) 

(sealed capillary), [aW^ —8° (c 1.0 in water). 

CH3S— CH2CH2CH— COOH 

I 
NH2 

Widely distributed. 

685 L-Ornithine, C5H12O2N0, colorless crystals, m.p. 140° (subl. 

120°), [alo'' +12° (c 6.5 in water). 

HoN— CH2— CH2— CH2— CH— COOH 

I 
NH2 

Widely distributed. 

686 Choline Sulfate, C5H13O4NS 

© 

(CHalsN— CH2CH2— O— SO3 

Aspergillus sydowi, Penicillium chrysogenum, lichens, 
yeasts 

Choline yields of 6000-7000 ^g. per gram of dry cell 
weight are available in certain Distillers' Dried Solubles. 

D. W. WooUey and W. H. Peterson, /. Biol. Chem. 122 213 
(1937). 

J. deFlines, J. Am. Chem. Soc. 77 1676 (1955). 

687 Imidazoleacetol (Hydrochloride), CgHgOoNaHCl, white needles, 

m.p. 171-174° (dec). 

O 

CH2— C— CH2OH 

/ 
©r=n 

HN:^ NH 

© 
CI 

Neuorspora crassa and E. coli mutants 



325 Amino Acids and Related Compounds 

Bruce N. Ames, Herschel K. Mitchell and Mary B. Mitchell, 
/. Am. Chem. Soc. 75 1015 (1953). 

688 L-Histidine, CoHoOsNa, colorless crystals, m.p. 287° (dec), 

[aln''' -39.7° "(c 1.13 in water). 

CH— C— CH2— CH—COOH 

I I I 

N NH NH, 

CH 

Claviceps purpurea (Fries) Tul. 

H. Heath and Jennifer Wildy, Biochem. J. 64 612 (1956). 

689 6-Diazo-5-oxo-L-norleucine (DON), C6H9O3N3, pale greenish 

yellow crystals, m.p. 145-155° (dec), [aW +21° (c 5.4 
in water). 

O 

N2CH—C—CH2—CH2— CH—COOH 

NH2 

An unclassified streptomycete 

Henry W. Dion, Salvatore A. Fusari, Zbigniew L. Jakubow- 
ski, John G. Zora and Quentin R. Bartz, /. Am. Chem.. Soc. 78 
3075 (1956). (Isolation and characterization) 

690 Imidazoleglycerol (Hydrochloride), CeHioOgNg-HCl, colorless 

crystals, m.p. 103° (dec), hW^ +13.3° (c 7.5 in water). 





CH- 


-CH— CH2 




/l 


1 1 


©r= 


n OH 


OH OH 


HN^ 


.NH 




e 






CI 







Neurospora crassa mutant 

Bruce N. Ames and Herschel K. Mitchell, /. Biol. Chem. 212 
687 (1955). 

691 L-Histidinol (Hydrochloride), CoH„ON3-2HCl, colorless crys- 
tals, m.p. 194° (dec). 





— 1 — 


CH2- 


-CH- 


-CH 


© 






1 © 


1 


HN^ 


NH 




NH3 


OH 


e ~^ 






e 




CI 






CI 





E. coli mutant 

Henry J. Vogel, Bernard D. Davis and Elizabeth S. Mingioli, 
/. Am. Chem. Soc. 73 1897 (1951). 



Pfizer Handbook of Microbial Metabolites 326 

692 L-Leucine, CcHj-iOMN, colorless crystals, m.p. ~295° (dec.) 

(sealed tube") (subl. from 140°), [a],,-' -11° (c 2.0 in 
water). 

CH3 

\ 

CH— CH2— CH— COOH 

/ I 

CH3 NH2 

Widely distributed. 

693 L-Isoleucine, CeHi.jO^N, colorless crystals, m.p. 284° (dec.) 

(subl. from 160°), [a],r" +11° (c 3.0 in water). 

CH3— CH2— CH— CH— COOH 

1 I 
CH3 NH, 

Widely distributed. 

694 L-a-Aminoadipic Acid, C(jHii04N, white crystals, m.p. 206° 

(dec). 

HOOC— CH,CH,CH,CHCOOH 

NH, 

Aspergillus oryzae 

Also a component of several antibiotics. 

Emmanuel Windsor, /. Biol. Chem. 192 595 (1951). 

695 L-Lysine, C,jH,40^.No. white needles, m.p. 224°. 

H,NCH,CH2CH2CH,CHCOOH 

NH.. 

Ustilago maydis PRL 1092 

The yield was 200-300 mg. per liter of free lysine in the 
broth as determined by a bioassay (not isolated). 

M. Richards and R. H. Raskins, Can. J. Microbiol. 3 543 
(1957). 

696 L-Arginine, C,jH,40oN4, colorless crystals (Dihydrate), m.p. 

245° (dec.) (browning above 200°), [a],,-" +13° (c 3.5 
in water). 

NH 

H N— C— NH— CH2— CH,— CH .— CH— COOH 

I 
NHs 

Widely distributed. 



3^7 Amino Acids and Related Compounds 

697 8-Oxy-L-lysine (a.f-Diamino-8-hydroxycaproic acid), C,.H,.,0:jNo. 

H .NCH,CHCH CH CHCOOH 

I I 

OH NH, 

Mycobacterium phlei 

Occurs bound in a phosphatide (yellow powder, m.p. 
180-190°), molecular weight about 16,000. It is the sole 
amino acid, and constitutes about 1 % of the phosphatide. 

M. Barbier and E. Lederer, Biochim. et Biophys. Acta 8 590 
(1952). 

698 Anthranilic Acid, C^H^O.N, leaflets, m.p. 144°. 

COOH 



NHa 

Corynebacterium diphtheriae 

Detected by paper chromatography. 

A. J. Woiwood and F. V. Lniggood, Intern. Congr. Biochem., 
Abstrs. of Communs. 1st Congr., Cambridge, England, 320 
(1949). 

Anthranilic acid has been isolated also from a pseu- 
domonas culture: 

Rokuro Takeda and 1. Nakanishi, /. Fermentation Technol. 
37 No. 2 (1959). 

It also accumulates in certain bacterial auxotrophs. 

699 p-Aminobenzoic Acid, C7H7O0N, yellowish red crystals, m.p. 
186°. 

H2N— / V-COOH 



Hansenula anomala, Mycotorula lipolytica 
Yields about 1 mg. per gram of dry cells. 
W. H. Peterson, "Yeasts in Feeding" Symposium, Milwaukee, 
1948. 

700 Trigonelline, CtH-O^.N, colorless crystals, m.p. (anhyd.) 218° 
(dec.) (Picrate) m.p. 205° (dec). 


COO 



© 



CH3 



Pfizer Handbook of Microbial Metabolites 328 

Polyporus sulfureiis 

P. H. List, Planta Med. 6 424 (1958). 

701 Homarine, C7H7O2N (Hydrochloride), m.p. 170-175° (dec.) 
(Picrate) m.p. 155-160°. 




c=o 

I e 

CH3 o 

Polyporus sulfureus 

P. H. List, Planta Med. 6 424 (1958). 

702 Stachydrine, C7H13O2N, white monohydrated crystals, m.p. 

(anhydr.) 235° (dec). 

CH2 CH2 

I I 

CH2 CH 

\ / \ 
N© C=0 

CH3 \oe 
CH3 

Aspergillus oryzae, other fungi (in small yields) 
R. Takata, J. Soc. Chem. Ind. Japan 32 155B (1929). 

703 2,6-Diaminopimelic Acid (Both l,l- and mesa forms occur nat- 

urally), C7H14O4N2, colorless needles, m.p. >305°. 

HOOC— CH— CHo— CHo— CHo— CH— COOH 

I I 

NH2 NHo 

Corynshacterium diphtheriae, Mycobacterium tubercu- 
losis. Bacillus anthracis, E. coli mutants 

Elizabeth Work, Biochem. J. 49 17 (1951). 

H. Smith, R. E. Strange and H. T. Zwartouw, Nature 178 
865 (1956). 

Lester E. Casida, Jr., U. S. Patent 2,771,396 (1956). 

704 /3-Methyllanthionine, C7H14O4N0S, [a]i>-° +37.6° (c 0.5 in 1 N 

hydrochloric acid). 

HOOC— CH— CH2— S— CH— CH— COOH 

I I I 

NH2 CHs NH2 

Yeast 

This isomer is not the same as the one isolated from 



329 Amino Acids and Related Compounds 

antibiotic hydrolysates. Desulfurization with Raney 
nickel yields L-alanine and D-a-amino-/2-butyric acid. 

Phyllis F. Downey and Simon Black, /. Biol. Chem. 228 171 
(1957). 

705 L-Phenylalanine, CgHnOoN, colorless crystals, m.p. 283° (dec.) 

(rapid heating), [aW^ —35° (c 2 in water). 

/ \— CH2— CH— COOH 

NH2 

Widely distributed. 

706 L-Tyrosine, C9H11O3N, colorless crystals, m.p. 342-344° (sealed 

capillary, preheated bath) (dec), [a]D" —10.6° (c 4 in 
1 N hydrochloric acid). 

HO—/ V-CHo— CH— COOH 

NH2 
Widely distributed. 

707 Hercynine (Histidine Betaine), C9H15O2N3, white crystals, no 

sharp m.p., forms mono- and dipicrates. 

HC— C— CH2— CH— COO ® 
N NH ©N(CH3)3 

H 

Amanita muscaria, Agaricus campestris, Boletus edulis 
Bull., Polyporus sulfureus 

Fr. Kutscher, Zentr. Physiol. 24 775 (1910). 

R. Engeland and F. Kutscher, ibid. 26 569 (1912). (Syn- 
thesis) 

Albert Kiing, Z. phijsiol. Chem. 91 241 (1914). 

708 Ergothioneine, C9H15O2N3S, colorless crystals, m.p. 290° (dec), 

[a]D-M16.5°. 

CH=C— CH2— CH— COO ® 

I I 1 

N NH N(CH3)3 

I 
SH 



Pfizer Handbook of Microbial Metabolites 330 

Claviceps purpurea (Fries), Tul. Coprinus comatus, 
Mycobacterium tuberculosis 

C. Tanret, J. pharm. chim. 30 145 (1909). 

H. Heath and Jennifer Wildy, Biochem. J. 64 612 (1956). 
(Biosynthesis) 

Paul Heinz List, Arch. Pharm. 290 517 (1957). 

Dorothy S. Genghof, Bad. Proc, 190 (1960). 

709 L-Tryptophan, CiiHi^O^,N2, colorless crystals, m.p. 289° (dec.) 
(rapid heating), [ix]i,-'^ —31.5 (c 1.0 in water). 

CH2— CH— COOH 



CO 



NH2 



H 
Widely distributed. 

710 Amino Acid from Lactarius helvus, CuHiyOaNo, colorless crys- 
tals, yellowing near 200° and darkening to 300°. Molecu- 
lar weight 251 by isothermal distillation. Adds 2 H^ and 
2 Bro. 

Partial structure: 

HOOC— CH CH— COOH 

NH,. ^ NH2 

C7H10 



C 

\^ 

containing C= and C=C 

/ 
C 

Lactarius helvus 

Ateushi Komamine and Artturi Virtanen, Acta Chem. Scaiid. 
13 2141 (1959). 

J. Casimir and A. I. Virtanen, ibid. 13 2139 (1959). (Iso- 
lation ) 

711 Elaiomycin, C,..iHo,;0;!N._,, pale yellow oil, [a],,"' +38.4° (c 2.8 in 
absolute ethanol). 

O OH 

T I 

CH3(CH2)5CH=CH— N=N— CH— CH— CH3 

CH0OCH3 
Streptomyces hepaticus 



33 1 Amino Acids and Related Compounds 

Theodore H. Haskell, Albert Ryder and Quentin R. Bartz, 
Antibiotics and Chcmotlierapy 4 141 (1954). (Isolation) 

John Ehrlich, Lucia E. Anderson, George L. Coffey, William 
H. Feldman, Myron W. Fisher, Arther B. Hillegas, Alfred G. 
Karlson, Mildred P. Knudsen, Jean K. Weston, Anne S. You- 
mans and Guy P. Youmans, ibid. 4 338 (1954). 

C. L. Stevens, B. T. Gillis, J. C. French and T. H. Haskell. 
;. Am. Chem. Soc. 78 3229 (1956). (Structure) 



17 



Polypeptides and Related Compounds 



Polypeptides are often intractable, difficultly crystallizable 
substances. The newer techniques of chromatography, end- 
group analysis and electrophoresis have facilitated their investi- 
gation. 

Most of the polypeptides and related compounds listed in this 
section are antibiotic isolates. Antibiosis may be a primary or 
only a secondary function of these materials. Polypeptides, of 
course, have hormonal and other functions in higher animals. 
Among microorganisms streptomycetes and bacteria have been 
the richest sources so far, perhaps in part because they have 
been examined more extensively for antibiotic activity than 
other microorganisms. 

Special types of polypeptides have been isolated from bac- 
terial cell walls by fragmentation with lysozyme or bacterio- 
phage. They also tend to accumulate when bacteria are in- 
hibited by certain antibiotics. Determination of their structures 
is beginning to elucidate the nature of the bacterial cell wall as 
well as the mode of action of the antibiotics involved. 

Some attention has been given to intracellular peptides, prin- 
cipally in connection with their role in protein synthesis. The 
fundamental process of polypeptide and protein biosynthesis is 
just beginning to yield some of its secrets. Before discussing it, 
some earlier work on simpler polypeptide biosynthesis will be 
reviewed. 

Glutathione is a widely distributed tripeptide which has a 
rapid metabolic turnover in yeast and also in mammalian tis- 
sues. Partly for this reason it has been suggested as an inter- 
mediate in protein biosynthesis, but because of its reversible 
oxidation-reduction properties, a respiratory role also has been 
proposed. In fact, it has not been proved satisfactorily that 
polypeptides serve as direct precursors for protein synthesis in 



333 Polypeptides and Related Compounds 

microorganisms, although strepogenins (glutamic acid contain- 
ing oligopeptides from the enzymic digests of certain proteins) 
stimulate the growth of some bacteria. There is evidence for 
the occurrence of independent uptake mechanisms for glycine 
and glycine peptides in Lactobacillus casei.^ 

Glutathione formation takes place in two separate reactions, 
each involving ATP : - 

( 1 ) L-Glutamic Acid + L-Cysteine + ATP^ 
L-y-Glutamylcysteine + ADP + H3PO4. 

(2) L-y-Glutamylcysteine + Glycine + ATP^ 
L-Glutathione + ADP + H3PO4. 

The biosynthesis of pantothenic acid probably proceeds as 
follows, the last step also being coupled with ATP cleavage, but 
with different products ■.^- * 

Ci unit from 
CH3 O tetrahydro- CH3 O 

\ II folic acid | || 

CH— C— COOH > HOCH2— C C— COOH 

CH3 CH3 \ 2H 

a-Ketoisovaleric Acid ^>i 

CH3 
I 
HOCH2— C CH— COOH 





CH3 OH 


Pyrophosphate + 


// Pantoic Acid 
/3-Alanine / 
CH, ^ ATP 
1 II 
-C CH— C— NH— CH2— CH2— COOH 


AMP + HOCH2- 




CH3 OH 




Pantothenic Acid 



^ Franklin R. Leach and Esmond E. Snell, Biochim. et Biophys. 
Acta 34 292 (1959). 

2 John E. Snoke and Konrad Bloch, /. Biol. Chem. 199 407 (1952); 
John E. Snoke, ibid. 213 813 (1955); John E. Snoke and Konrad 
Bloch, ibid. 213 825 (1955); Stanley Mandeles and Konrad Bloch, 
ibid. 214 639 (1955). 

3 Werner K. Maas and Henry J. Vogel, /. Bacteriol. 65 388 (1953); 
M. Purko, W. O. Nelson and W. A. Wood, /. Biol. Chem. 207 51 
(1954); E. Nelson Mcintosh, M. Purko and W. A. Wood, ibid., 228 
499 (1957). 

* Werner K. Maas, /. Biol. Chem. 198 23 (1952); Akira Matsuyama, 
Bull. Agr. Chem. Soc. (Japan) 21 47 (1957) and earlier papers in 
this series; Herbert S. Ginoza and Robert A. Altenbern, Arch. 
Biochem. and Biophys. 56 537 (1955). 



Pfizer Handbook of Microbial Metabolites 



334 



It appears that an adenylic acid-pantoate complex is the inter- 
mediate which couples with the enzyme. 



NHj 



sXn 




o o 

T T I 

CH,— O— P— O— P— O— CH2— C 



OH 



OH 



CH3 OH O 

I I II 

CH— C— NH— CH2— CH,— COOH 

CH3 



Pantothenylcysteine is a precursor of pantetheine in Lactobacil- 
lus helveticus.^ 

The red actinomycins were the first antibiotics isolated crys- 
talline from actinomycetes/' In the ensuing 20 years about a 
dozen named species of streptomycetes have been found to pro- 
duce actinomycins, and probably many other isolates have gone 
unreported. 



TABLE I 
Chronological List of Microorganisms Reported to Produce Actinomycins* 



Year reported 


Name given to 
microorganism 


Actinomycin 
complexf 


1941 


Sfrepfomyces ant/biot/cus 


A 


1946 


Non-chromogenic species 


A 


1947 


S. flovus 


A (J) 


1948 


S. parvus 


A 




S. flavovirens 


— 


1949 


Streptomyces sp. 


B 




S. chrysomallus 


C 


1951 


S. flaveolus 


A (J) 




Micromonospora globosa 


— 


1952 


Streptomyces sp. 


X 



5 Gene M. Brown, /. Biol. Chem. 226 651 (1957). 
« S. A. Waksman and H. B. Woodruff, Proc. Soc. Exp. Biol. 45 609 
(1940). 



335 



Polypeptides and Related Compounds 



TABLE 1 -Continued 



Year reported 


Name given to 
microorganism 


Actinomycin 
complexf 


1954 


S. 


flovus 




X(B) 




S. 


flavus 




X 




S. 


aniibioticus 




X 




S. 


flavus-parvus 




X(B) 




S. 


parvullus 




D 




S. 


cellulosae 




— 




S. 


michiganensis 




X 




S. 


anfibioficus 




M 


1956 


Strepfomyces sp. 




E 




Sfrepfomyces sp. 




F 


1958 


S. 


fradiae 




Z, X 



* By courtesy of Dr. H. Boyd Woodruff, Merck, Shorpe and Dohme, and the New York 
Academy of Science. 

t See the discussion of nomenclature preceding the actinomycin entries. 

Often these polypeptide pigments occur in closely related com- 
plexes, the individual members varying only by slight changes 
in the side-chains. 

The chromophore, actinocinin, resembles that of the ommo- 
chromes, a group of insect pigments studied by Butenandt,' and 
it is similar to the pigment cinnabarin from a higher fungus. 

COOH 
NH, 



COOH 





CH3 CH3 

Actinocinin 



^o--^=^\ 
o 

Xanthommatin 

^ Adolph Butenandt, Ulrich Schiedt, Ernst Biekert and Pierre 
Kornmann, Ann. 586 217 (1954); Adolph Butenandt, Ulrich Schiedt 
and Ernst Biekert, Ann. 586 229, 588 106 (1954); Adolph Butenandt, 
Ulrich Schiedt, Ernst Biekert and R. Jan. T. Cromartie, Ann. 590 75 
(1954). (Structure) 



Pfizer Handbook of Microbial Metabolites 



336 




CH3 CH3 

Actinomycin C3 



The dual nature of the actinomycin molecules makes it rather 
obvious that they must be formed by condensation of two similar 
halves. Butenandt showed that xanthommatin was derived 
from tryptophan by feeding experiments with the labeled amino 
acid. Similarly Katz has shown® that actinocinin is derived 
from tryptophan. Thus the entire actinomycin molecule is 
composed of amino acid derivatives. In the case of xanthom- 
matin the intermediate is kynurenine, a known degradation 
product of tryptophan. Kynurenine may also be an intermedi- 
ate in actinocinin biosynthesis, although the assumed inter- 
mediate, 3-oxy-4-methyl-anthranilic acid might equally well 
arise through a variation in the biosynthetic route to tryptophan. 

^ Edward Katz, N. Y. Acad. Sci. Conference on Actinomycins, 
March 31 to April 1, 1960. (Unpublished) 



337 



Polypeptides and Related Compounds 



CXJ 



CH2— CH— COOH 

1 
NH, 




Tryptophan 



CO— CHo— CH— COOH 
NHo 

NHo 

Kynurenine 

CO— CH.— CH— COOH 

I 
NH, 




3-Oxykynurenine 

It is likely that the peptide side-chain is attached before conden- 
sation to form the chromophore. 



COOR 



COOR 



COOR 




OH 

CH3 

3-Oxy-4-methyl- 

anthranilic Acid 

(R = Polypeptide 

Moiety) 



Actinomycins 



It is interesting that methyltryptophans (a,5,6-methyls) are 
inhibitory to actinomycin production. Methionine is probably 
the donor of the methyl groups in N-methylvaline and sarcosine.^ 

In two streptomycete species D-valine inhibits actinomycin 
synthesis while L-valine stimulates it, although D-vahne is the 
isomer present in the side-chains.^ This behavior is reminiscent 
of the results of similar earlier experiments with penicillin and 
with valinomycin. 

Schmidt-Kastner found that addition of a large quantity of 
sarcosine to the medium caused replacement of part or all of the 
side-chain proline by sarcosine. Analogously, addition of isoleu- 
cine caused replacement of N-methylvaline by N-methyhsoleu- 
cine.'' Since then many new actinomycins have been prepared 
by addition of various amino acids to the medium. Even 

^ G. Schmidt-Kastner, Naturwissenschaften 43 131 (1956). 



Pfizer Handbook of Microbial Metabolites 338 

pipecolic acid can be incorporated.'" It should be noted that 
certain amino acid analogues can be incorporated into enzymes 
and other proteins without impairing their normal functions." 

Professor Hans Brockmann and his collaborators at Gottingen, 
who were primarily responsible for determining the structure of 
the first well-characterized actinomycin, actinomycin C^,'- have 
succeeded in synthesizing this substance' ' and it should be pos- 
sible now to prepare an even wider variety of actinomycins. 

Valinomycin, shown opposite, can be hydrolyzed to its con- 
stituents: L-valine, D-valine, D-a-hydroxyisovaleric acid and 
L-lactic acid. It has been found'* that L-valine- l-C* in the 
medium was incorporated to an equal extent into the D-valyl 
and L-valyl portions of the molecule, to a lesser extent into the 
D-a-hydroxyisovaleric acid, and not at all into the lactic acid. 
D-Valine-l-C* was incorporated only to a slight extent. Similar 
results have been obtained in studies on the origin of the 
D-valine moieties in penicillin and in actinomycin. 

The co-occurrence of valine with the biosynthetically related 
-z-hydroxyisovaleric acid in several polypeptides is noteworthy. 
Also conjugated with certain polypeptides are 6-methyloctanoic 

CHs— CH2— CH— CH2— CH.— CH,— CH,— COOH 

I 
CH3 

6-Methyloctanoic Acid 

CH3— CH2— CH,— CHo— CH.— CH.— CH>— CH— CHo— COOH 

I 
OH 

3-Oxydecanoic Acid 

acid and 3-oxydecanoic acid. The latter substance has been 
found conjugated with bacterial carbohydrates too. 

10 Edward Katz and WilHam Goss, Nature 182 1668 (1958); S. A. 
Waksman, E. Katz, W. A. Goss, L. H. Pugh, M. Solvtorowsky, and 
N. A. Auerbach, Science 129 1290 (1959); E. Katz and W. A. Goss, 
Biochem. J. 73 458 (1959); A. W. Johnson and A. B. Mauger, ibid. 
73 535 (1959); WiUiam A. Goss and Edward Katz, Antibiotics and 
Chemotherapy 10 221 (1960). 

" E.g., Akira Yoshida and Mekoto Yamasaki, Biochim. et Biophys. 
Acta 34 158 (1959). 

1- H. Brockmann, G. Bohnsack, B. Franck, H. Grone, H. Muxfeldt 
and C. Siiling, Angeiv. Chem. 68 70 (1956); H. Brockmann, N. 
Grubhofer, H. Kalbe and W. Kass, Chem. Ber. 84 260 (1951); H. 
Brockmann, Angeiv. Chem. 66 1 (1954); H. Brockmann and B. 
Franck, ibid. 68 70 (1956) and other papers in this series. 

1-^ H. Brockmann, W. Sunderkotter, K. W. Ohly and P. Boldt, 
NatUTivissenschaften 47 230 (I960); H. Brockmann and H. Lackner, 
ibid. 47 320 (1960). 

"J. C. MacDonald, Can. J. Microbiol. 6 27 (1960). 



339 



Polypeptides and Related Compounds 



CHa 



\ 



CH 



CHs 





CH3 




II 
-C— 


CH3 




\ 


CH— CH 
~NH^^ / 

i-Valine CH 

X 

e 




L-Lactic 
Acid 




CH D-Valine 







NH 



D-a-Hydroxy- CH 
isovaleric Acid 



CH 



0=C 



CHa 



CH3 



CH3 



CH3 



c=o 



CH' 



D-a-Hydroxy- 
-CH isovaleric 
\ Acid 



NH 



D-Vaiine CH /"' 



CH 



./ 



^L-Va!ine 
CH^ 

/ 



CH 

/ \ 
CHa CH3 



NH 



L-Lactic X O 

Acid^.^0 
-- C -CH'"^ 

" CH 

Vaiinomycin 



\ 



CHs 



CH2 



/ 



NH. 

I 

CH, CH3 CH3 

I \ / 

CHa CHa CHo CH 

\ / 1 I 

CH CH2 CHo CH2 

I I I I 

CO— NHCHCO NHCHCO NHCHCO NHCHCO— N CH2 

I (l) (l) (l) (d) 

CHo— CH 

(l) (l) CH— CHo 

(d) (l) (l) (l) I 
CH2— N COCHNH COCHNH COCHNH COCHNH — CO 

CH2 



CH, 



CH2 

1 

CH 

/ \ 
CHa CHa 


CH2 
CH2 
CH2 
NH2 


CH 

/ \ 
CHa CH 


Gramicidin S 





Pfizer Handbook of Microbial Metabolites 



340 



The biosynthesis of gramicidin S has been studied. ^^ The 
conclusions were: (a) The five amino acids of the cyclic de- 
capeptide pass through a number of intermediates before or 
during incorporation, (b) Final formation of gramicidin S is a 
simple reaction not requiring free amino acids which occurs 
readily in cell-free suspensions, (c) Three peptides were iso- 
lated containing fragments of the amino acid sequences of the 
antibiotic. These may or may not have been intermediates. 

It is possible to extract intracellular peptides with suitable 
solvents. This has been done with mammalian pituitary tis- 
gyg 16. 17 ^fjj plant seeds^^ and with yeast^" and bacteria.-"' ^^ In 
all cases cited care was taken to obviate contamination by frag- 
ments of proteolysis. There is some indication that yields are 
higher from rapidly growing bacteria than from resting cells. 

The intracellular peptides of the torula yeast studied were 
found to be predominantly acidic with glutamic acid the princi- 
pal amino acid. About 40 peptides were purified in adequate 
quantity to permit hydrolysis and identification of constituent 
amino acids. These are tabulated below (x indicates an un- 
identified ninhydrin-positive substance ) : 



TABLE II 
Some Intracellular Peptides of Torula Yeast 



Peptide 
No. 


Amino acid content 


Peptide 
No. 


Amino acid content 


1 


Glu, x-6, Gly, Alo, Asp, Arg, 
Vol 


21 
22 


Glo (Gly, x-3, Ala) 

Glu, Gly, Cys (Glutathione) 



15 J. M. Barry and Elizabeth Ishihara, Nature 181 1274 (1958). 

I*' T. Winnick, R. E. Winnick, R. Acher and C. Fromageot, Biochim. 
et Biophys. Acta 18 488 (1955). 

" L. K. Ramachandran and T. Winnick, ibid. 23 533 (1957). 

1^ H. Borriss and G. Schneider, Naturwissenschaften 42 103 
(1955). 

"F. Turba and H. Esser, Biochem. Z. 327 93 (1956). 

2° G. E. Connell and R. W. Watson, Biochim. et Biophys. Acta 24 
226 (1957). 

21 R. B. Roberts, P. H. Abelson, D. C. Cowie, E. T. Bolton and R. J. 
Britten, "Studies of Biosynthesis in E. coli," Carnegie Institute, Wash- 
ington, D. €., 1955. 



341 



Polypeptides and Related Compounds 



TABLE ll-Continued 



Peptide 
No. 


Amino acid content 


Peptide 
No. 


Amino acid content 


2 
3 


Glu, Gly, Ala, Asp, Ser, Vol, x-7, 

Arg 

Glu, Gly, Asp, Ala, Thr? x-6. 


23 
24 


Glu, Gly, Ala, His, Arg or. Cys, 

x-6. Asp, Lys, Vol 

Glu, Gly, Ala, x-7, x-11. Asp, 


4 


Arg? 

Glu, Gly, Ala, Thr? Asp, Arg, His, 


25 


Ser, Leu, Vol, Arg, Lys 

Gly, Glu, x-6, Ser, Ala, His, Vol, 




Vol, x-5 




Leu, Asp 


5 
6 


Glu, Gly, Ala, His, Asp, x-4 
Glu, Gly, Ala, x-5, Asp, Arg, Vol, 


26 


Glu, Gly, x-4, Ala, His, Lys, 
Leu 


7 


His 

Glu, Gly, His, Ala, x-5. Asp, Arg, 


27 
28 


Gly, Ala, Glu, x-4. Vol, Arg 
Glu, Gly, x-11, Ser, Ala, Arg, 


8 


Vol 

Glu, Gly, Ala, Asp, x-4. His, 


29 


Thr, X-7, Asp, Vol, Lys 

Glu, Gly, Ser, Ala, x-11. Asp, 


9 


Arg 

Asp, Gly, Glu, Ala, x-5. Vol, 


30 


Thr, Vol, Lys, Arg, Leu 

Glu, Gly, Ser, Ala, x-8. Asp, Thr, 


10 


Arg 

Ala, Gly, Glu, x-5, x-6. Vol 


31 


x-11. Vol, Leu, Arg 

Gly, Glu, x-4, Ser, Ala, Asp, 


11 


x-4, x-7, x-5, Gly, Glu, Ala, 




Leu 


12 


Asp 

Asp (Gly, Glu, Ala, x-5, x-6) 


32 

33 


Gly, x-5, Glu, Ala, Asp, Arg 
Glu, Gly, x-6, Ala, a-But, Leu 


13 


Glu, Gly, Ala, Asp, x-5 


34 


x-3, Ala, Gly, Glu, x-7, Ser, Asp, 


14 


Glu, Gly, Ala, x-5, x-8, Ser, Asp, 




Vol, Arg, Leu 


15 


Vol, Arg? 

Ala, Gly, Glu, x-4, x-6. Asp 


35 


Gly, Glu, Ala, x-7, Arg, Asp, 
Vol, Leu 


16 


Glu, Gly, His, Ala, Cys, x-4 


36 


Arg? x-3, Gly, Glu, Ser., Ala, 


17 


Glu, Gly, Cys, x-10, Ala, Ser, 




x-8, x-12. Asp, Thr, Vol, Leu 




x-6. Asp, Arg, Vol, Leu 


37 


Gly, Glu, Ser, Ala, Asp, x-5, Arg, 


18 


Glu, Gly, x-9, Ser, Ala, Asp, Thr, 




x-9, Thr? Vol 




Cys? Arg, x-5 


38 


Gly, Ser, Gly, Ala, Asp, Vol, 


19 


Gly, Glu, x-6, Ala, Ser, Asp, Vol, 




x-6, Thr, Arg, Lys, His, x-12 




Leu, His 


39 


Gly, Glu, Ala, x-6. Leu, Vol, Thr, 


20 


Glu, Gly, Ala, Asp, x-7, Ser, Tyr, 




Asp, x-11. His, Lys 




Vol, Leu, His 


40 


Gly, Glu, x-5, Ala, a-But, Vol 



In a similar study with the use of E. coli ten intracellular 
peptides were purified in sufficient amounts to allow amino acid 
determination.-- In this case the N-terminal amino acids were 

^^ D. Griinberger, Jifina Cerna and F. Sorm, Experientia 16 54 

(1960). 



Pfizer Handbook of Microbial Metabolites 



342 



distinguished by formation of their dinitrophenyl derivatives. 
The results are shown in the following table: 



TABLE III 
Some Infraceltular Peptides of Escherichia co/i 



Peptide No. 


Terminal amino acid 


(Other amino acids) 


1 


Glu 


(Cys, Gly, Lys) 


2 


Glu 


(Ala, Cys, Gly, Lys) 


3 


Asp 


(Cys, Gly, Lys) 


4 


Lys 


(Ala, Arg, Asp, Cys, Gly, Glu, Ser) 


5 


Asp 


(Arg, Gly, Glu, 7-NH.But, Lys, Vol) 


6 


Ser 


(Asp, Gly, Lys) 


7 


Ala 


(Asp, Lys) 


8 


Glu 


(Ala, Asp, Cys, Gly, Lys, Leu, Vol) 


9 


Glu 


(Ala, Asp, Lys, Cys, Gly) 


10 


Glu 


(Cys, Gly) 



It has been reported that gram-negative bacteria contain 
much less intracellular free ninhydrin-positive substances than 
do gram-positive ones.-^ 

A basic polypeptide has been extracted from dried cells of the 
human strain of Mycobactenum tuberculosis H.jyR,, purified, 
crystallized and quantitatively analyzed for amino acid con- 
stituents.-' The pure peptide showed activity in the tuberculin 
test at least equal to that of standard old tuberculin. The amino 
acid content was as follows, subscripts indicating number of 
moles : 

Argio HiSi Lyso Phe, Tyr, Leu..^ Ileu^. Val- 
Alag Glye Glus Pro- Ser^ Thr.{ Asp,, Tryi 

The molecular weight was calculated to be 7180. 

Certain polypeptides accumulate in E. coli cells grown in the 
presence of chloramphenicol (a protein synthesis inhibitor). 
Two of these have been isolated and purified.-^ 

-' Yuichi Yamamura, Seisi Morizawa, Atsushi Tanaka, and Kenji 
Shojima, Proc. Jap. Acad. Sci. 35 295 (1959); Seisi Morizawa, Atsushi 
Tanaka, Kenji Shojima and Yuichi Yamamura, Biochivi. et Biophys. 
Acta 38 252 (1960). 

-' F. Sorm and Jifina Cerna, Collection Czechoslov. Chem. Com- 
muTi. 25 565 (1960). 



343 



Poln^eptklesandRelated Compounds 

Synthesis of the cell wall mucopeptides of staphjd^^o^^ 
unaffected by chloramphenicol, but inhibited (at least indi- 
rectly) by penicillin, bacitracin, cycloserine, novobiocin and 
gentian violet. None of these inhibits protein synthesis 

Penicillin-inhibited Staphylococcus aureus accumulates three 
closely related uridine nucleotides.- One of these has been as- 
signed the structure : '■"'■ -'■ -^ 



\/N 




^N-Acetylmuramic Acid 



CH— CH.— CHo— COOH 

NH2 

I 
NH CH, 

D-Ala D-Ala | I 

HOOC-CH-NH-C-CH-NH-C-CH-CH2-CH,-CH2 
I II I II L-Lys 

CH3 O CH3 O 

This fragment may be the repeating unit of an activated cell 
wall precursor, since the ratio of muramic acid: alanine glu- 
tamic acid:lysine is 1 :3:1 :1, the same ratio found in lysozyme 
digests of whole bacteria. In E. coli and Corynebacterium 
diphthenae the lysine in the peptide chain is replaced by its 
biosynthetic precursor, meso-diaminopimelic acid. 

^^ J. T. Park and N. J. Johnson, /. Biol. Chem. 179 585 (1949). 

J. T. Park and J. L. Strominger, Science 125 99 (1957). 
2^ J. L. Strominger, /. Biol. Chem. 234 1520 (1959). 
'-ndem.. Federation Proc. 18 334 (1959); Eijl Ito and Jack L. 
Strommger, J. Biol. Chem. 235 PC5 (1960). 



Pfizer Handbook of Microbial Metabolites 344 

There is increasing evidence that the antibiotics mentioned, 
lysozyme and bacteriophages, all bring about a similar result, 
the accumulation or liberation of a fundamental cell waU unit 
such as the one shown. Lysozyme and bacteriophages are able 
to liberate the unit from pre-formed walls, while the antibiotics 
merely block wall synthesis. Also, the unit obtained by lyso- 
zyme or phage action seems to contain glucosamine as well as 
muramic acid, and sometimes diaminopimelic acid (a lysine 
precursor) rather than lysine. There is evidence that N-acetyl- 
D-glucosamine is a direct precursor of muramic acid. 

COOH 

I 

c=o 

I 

CH2 

I 

CH— OH 

I 
H2N— CH 

! 

HO— CH 

1 
HC— OH 

I 
HC— OH 

I 
CH2OH 

Neuraminic Acid 

Several neuraminopeptides have been isolated from an E. coli 
mutant culture, and one of these has been purified.-^ It is com- 
posed of N-acetylneuraminic acid, glucosamine, alanine, lysine 
and glutamic, acid. '^" 

A model of cell wall structure in gram-positive bacteria has 
been postulated :^^ 

P P 

/ . . / . 

M — r^G >M— ^-*G >M -^ G 

t * M = Muramic Acid 

G G = N-Acetyl-D-glucosamine 

...T... P = Peptide Moiety 

P P ' *' • = Lysozyme Action 

/ . / : • 

M — r-^G >M — j->G >M 

« 

29 P. J. O'Brien and F. Zilliken, Biochim. et Biophys. Acta 31 543 
(1959). 

^° E. Kean, Dissertation. (In press) 

31 Friedrich Zilliken, Federation Proc. 18 966-973 (1959). 



345 Polypeptides and Related Compounds 

The spine is composed of alternating muramic acid and N- 
acetylglucosamine units with branching to adjacent chains from 
muramic acid, the latter bearing the peptide chain. Penicillin 
(and perhaps the other antibiotics mentioned) prevents incor- 
poration of M-P units, and cycloserine prevents incorporation of 
the terminal two D-alanine units into the side-chain. There is 
evidence that the dipeptide D-alanyl-D-alanine is preformed be- 
fore attachment to the peptide chain. 

A review of the chemistry of bacterial cell walls has been 
published. -^^ 

The newer general theory of polypeptide and protein synthesis 
can be sketched in only briefly here.^- It is thought that the 
DNA of the cell nucleus lays out the pattern for replication of 
the ribosomal RNA, and this pattern is characteristic of each 
genus, species and type of organism. The ribosomal RNA in 
turn serves as the template for protein construction. Smaller, 
more soluble molecules, which seem to be RNA fragments end- 
ing in the nucleotide adenylic acid, attach themselves at this end 
to individual amino acids. This attachment requires an enzyme 
specific for each of the 20 or more amino acids plus ATP. There 
is also a different transfer RNA molecule for each amino acid. 
These activated amino acids can be isolated and purified. In 
this form the amino acid is able to fit into the proper place on 
the RNA template, probably due to the unique geometry of a 
short sequence of nucleotides in the chain. Once attached to 
RNA, condensation of the amino acids to form polypeptides or 
proteins is facilitated by the favorable arrangement and proxim- 
ity of the reacting groups. This scheme is believed to be quite 
general in metabolism. 

A more specific discussion by E. F. Gale of current knowledge 
about the incorporation of amino acids into bacterial proteins 
and polypeptides has been published. ^^ It is obvious that con- 
siderable differences must exist between mechanisms of polypep- 
tide synthesis in microbial and mammalian metabolism in view 
of the D-amino acids and other abnormal amino acids which oc- 
cur in microbial polypeptides. It is apparently these differences 

32 Robert B. Loftfield, Prog. Biophys., Biophys. Chem. No. 8 348 
(1957); F. H. C. Crick, Symposia of the Society for Exp. Biol. No. 12 
138 (1958); Mahlon B. Hoagland, Scientific American 201 55 (1959); 
Alton Meister, Rev. Mod. Phys. 31 210-220 (1959); Leo Szilard, Proc. 
Nat. Acad. Sci. U. S. 46 277 (1960). 

33 "CIBA Lectures in Microbial Chemistry," E. F. Gale, Synthesis 
and organization in the bacterial cell, John Wiley and Sons, New 
York, 1959, 106 pp. 



Pfizer Handbook of Microbial Metabolites 346 

which are exploited by some of the more successful antibiotics. 
Certain compounds listed elsewhere might have been classed 
as polypeptides. Examples are: penicillins, gliotoxin, certain 
ergot alkaloids, various diketopiperazines, netropsin, amicetin, 
Vitamin B^. conjugate and other fohc acids. 

712 DL-Fumarylyl Alanine (Fumaromono-D,L-alanide), CYHgOgN, col- 

orless needles, m.p. 229° (dec). 

HOOC— CH=CH— CO— NH— CH— COOH 

I 

CH3 
Penicillium resticnlosum 

John Howard Birkinshaw, Harold Raistrick and George 
Smith, Biochem. J. 36 829 (1942). 

713 Nocardamin, CgHi602N2, white needles, m.p. 184°, no optical 

activity. 

O 





II 


OH 




. / 


CH2 




^N 


/ 




\ 


CH2 




CH2 


NH 

\ 




CH2 


CH2 




CH2 



^CHa^^ 

Actinomyces buchanan 

A. StoU, J. Renz and A. Brack, Helv. Chim. Acta 34 862 
(1951). 

R. F.jC. Brown and G. Biichi, unpublished. (Revised struc- 
ture ) 

714 N-Succinyl-L-glutamic Acid, C9H13O7N (Monohydrate), colorless 
hygroscopic crystals, m.p. 62-64°, [«]»-" —11° (c 1.07 in 
water). 

HOOC— CH2—CH2—CH— COOH 

NH— C— CH2— CH2— COOH 

II 
O 

Bacillus megatherium 

This substance appears during the sporulating phase 

before the appearance of dipicolinic acid. 



347 Polypeptides and Related Compounds 

Jean Paul Aubert, Jacqueline Millet, Elisabeth Pineau and 
Gerard Milhaud, Compt. rend. 249 1956 (1959). 

715 Lycomarasmine, CjiHi-.O-N.., white powder, m.p. 227-229 (dec). 
Tentative structure : 

H,N-CO-CH, CH3 

HOOC— CH— NH— CO— CH,— NH— C— OH 

COOH 

Fusariinn lycopersici Sacc. 

This is the toxin of fusarium wilt. A second compound, 
C9H1.O-N0, white powder, m.p. 273-276° (dec), has been 
isolated from the mother Hquors. It is produced in up to 
three times the yield of lycomarasmine, but is biologically 
inactive. It is also produced (with evolution of ammonia) 
by boiling lycomarasmine with water. 

The yield of lycomarasmine in the initial isolation was 
80-110 mg. per liter. 

There is still some dissatisfaction with this structure. 

PI. A. Plattner and N. Clauson-Kaas, Helv. Chim. Acta 28 
188 (1945). (Isolation) 

D. W. WooUey, ;. Biol. Chem. 176 1291 (1948). (Struc- 
ture) 

M. Brenner, R. Tamm and P. Quitt, Helv. Chim. Acta 41 
763 (1958). (Criticism of structure) 

716 d-Pantothenic Acid, C^Hi-OsN, viscous oil, [aji,-' +37.5° (in wa- 
ter). 

CH3 O 

HOCH2— C— CH— C— NH— CH— CHCOOH 

I 1 
CH3OH 

Penicillin liquors yield 600-800 /xg. per gram of dry 
cell weight. 

Yeasts contain 150-300 fxg. per gram of dry cell weight. 

D. W. WooUey, /. Am. Chem. Soc. 62 2251 (1940). (Syn- 
thesis ) 

Leland A. Underkofier and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
1954 Vol. II, J. M. Van Lanen, Production of vitamins other 
than riboflavin, chap. 6, pp. 191-216. 



Pfizer Handbook of Microbial Metabolites 348 

717 Toxin of tobacco wild-fire disease, C10H16O6N2. 

Probable structure: 

NH2 OH 

I I 

HOOC— CH— CH.— CHo— CH— CH— C=0 

I I 
NH O 

1 I 
0=C CH— CH3 

Pseudomonas tabaci 

The toxin can by hydrolyzed to lactic acid and the 
amino acid, tabtoxinin, C7H14O5N2, (a,e-diamino-/3-hy- 
droxypimelic acid ) : 

HOOC— CH— CH2— CHo— CH— CH— COOH 

I II 

NH2 OH NH2 

D. W. Woolley, G. Schaffner and Armin C. Braun, J. Biol. 
Chem. 198 807 (1952). (Isolation) 

Idem., ibid. 215 485 (1955). (Structure) 

718 Glutathione (Glutamylcysteinylglycine) CiyHi^OoNgS, colorless 

crystals, m.p. 190-192° (dec). Unstable. [a]Hg.'' ' -9.4° 
in water, —85° in 10% hydrochloric acid. 

O O 

II ■ II 

HOOC— CH— CH2— CH2— C— NH— CH— C— NH— CH2— COOH 

I I 

NH2 CH2SH 

Yeasts 

F. G. Hopkins, Biochem. J. 15 286 (1921). (Isolation) 
Charles Robert Harington and Thomas Hobson Mead, ibid. 
29 1602 '(1935). (Synthesis) 

719 N-Succinyl-L-diaminopimelic Acid, CnHigOjN^. 

NH2 

HOOC— CH—CH,CH2CH2—CH— COOH 

NH— C— CH2CH2COOH 

II 
O 

Charles Gilvarg, Biochim. et Biophys. Acta 24 216 (1957). 

Lactobacillus bulgaricus Factor (Pantetheine and Pantethine), 
C11H00O4N0S and C22H4.O8N4S.. 



349 Polypeptides and Related Compounds 

720 Pantetheine: Colorless, hygroscopic, amorphous powder, [a]i,^" 

+ 12.9° (in water). 

CHs 

I OH 

HOCH,— C— CH— CO— NH— CH,— CH2— CO— NH— CH,— CH,— SH 

CH3 

721 Pantethine: viscous oil. 

CH3 
I 
IHOCH2— C CH— CO— NH— CH2— CH2— CO— NH— CH2— CH2— S— 12 

I I 
CH3 OH 

Yeasts, Ashbya gossypi, many other microorganisms 

William L. Williams, E. Hoff-Jorgensen and Esmond E. 
SneU, /. Biol. Chem. 177 933 (1949). 

Esmond E. Snell, Gene M. Brown, Vincent J. Peters, Jean A. 
Craig, E. L. Wittle, J. A. Moore, V. M. McGlohon and O. D. 
Bird, J. Am. Chem. Soc. 72 5349 (1950). 

Vincent J. Peters, Gene M. Brown, William L. Williams and 
Esmond E. SneU, ibid. 75 1688 (1953). 

Gene M. Brown and Esmond E. Snell, ibid. 75 1691 (1953). 

722 Glutathione-Cysteine Disulfide, C13H22O8N4S0. 

H2N— CH— CH2— CH,— CO— NH— CH— CO— NH— CH2— COOH 

I 1 

COOH CH2 

I 

S— S— CH>— CH— COOH 

I 
NH2 

Saccharomyces cerevisiae 

Glutathione itself occurs in yeasts. The disulfide above 
was not isolated. 

Arthur H. Livermore and Edward C. Muecke, Nature 173 
265 (1954). 

723 Serratamic Acid, CjaHo.-.OsN, colorless crystals, m.p. 138° (dec.), 

[(x]v-° -10.2° (c 5.0 in ethanol). 

O CH.OH 

II I 

CH3(CH2)6CHCH2C— NH— CH 

I I 

OH COOH 



Pfizer Handbook of Microbial Metabolites 350 

Serratia species 

Yields as high as 8 g. per liter have been reported. 
Hydrolysis gives L-serine and ( — )-3-oxydecanoic acid. 
The latter acid also has been found in conjugation with 
rhamnose and with other amino acids (see Viscosin). 

N. J. Cartwright, Biochem. J. 60 238 (1955). 

Idem., ibid. 67 663 (1957). (Structure) 

724 8-(a-Aminoadipyl) cysteinylvaline, C14H25O6N3S. 

CH3 CH3 

\ / 
CH 

O CH,— SH 

HOOC— CH— CHo— CHo— CHo— C— NH— CH— C— NH— CH— COOH 

NH2 O 

Fenicillium chrysogenum 

This tripeptide was isolated from the mycelium of the 
penicillin-producing mold. It may be a penicillin precur- 
sor since cyclization in the proper way would yield syn- 
nematin-B (cephalosporin-N) which differs from penicil- 
lin only in its side-chain. Synnematin never has been 
isolated from P. chrysogenum, however. 

H. R. V. Arnstein, D. Morris and E. J. Toins, Biochim. et 
Biophys. Acta 35 561 (1959). 

725 Alazopeptin, Ci.^Ho^OeN^, no definite m.p., [a]i,-' +9.5° (c 4.7 in 

water). 

A peptide containing 1 mole of a-alanine and 2 moles 
of 6-diazo-5-oxoaminohexanoic acid (DON) or an isomer. 

Streptomyces griseoplanus 

S. E. DeVoe, N. E. Rigler, A. J. Shay, J. H. Martin, T. C. 
Boyd, E. J. Backus, J. H. Mowat and N. Bohonos, "Antibiotics 
Annual 1956-1957," Medical Encyclopedia, Inc., New York, 
p. 730. 

726 Antibiotic I.C.I. 13,959. 



Acid hydrolysis yielded: 
a-Aminoisobutyric Acid 



CH3 COOH 



CH3 NH2 



35 1 Polypeptides and Related Compounds 

j3-Hydroxyleucine 

CH3 

\ 

CH— CH— CH— COOH 

/ I ! 

CH3 OH NH> 

as well as L-leucine, ^-alanine and y-methylproline. The 
ytJ-hydroxyleucine, which had not been reported previously 
as a natural product, has either the d- or L-t/zreo but not 
the erythro configuration. 

A Paecilomyces strain 

G. W. Kenner and R. C. Sheppard, Nature 181 48 (1958). 

727 Viomycin (Vinactin A, Vinactane, Celiomycin, Viocin), 

CiT-isHsi-sjOsNg, Sulfate: m.p. (anhydrous) 252° (dec.) 
(hydrated) 280° (dec), [a],,--' -32° (c 1 in water). Rota- 
tion varies with pH. 

A strongly basic polypeptide. The following compo- 
nents have been identified: a,/^-diaminopropionic acid, /?- 
lysine, L-serine and a guanidino compound. Salts are 
neutral. 

Streptomyces fioridae, S. puniceus, S. vinaceus 

A. C. Finlay, G. L. Hobby, F. Hochstein, T. M. Lees, T. F. 
Lenert, J. A. Means, S. Y. P'An, P. P. Regna, J. B. Routlen, 
B. A. Sobin, K. B. Tate and J. H. Kane, Am. Rev. Tuberc. 63 1 
(1951). 

Quentin R. Bartz, John Ehrllch, James D. Mold, Mildred A. 
Penner and Robert M. Smith, ibid. 63 4 (1951). 

Theodore H. Haskell, Salvatore A. Fusari, Roger P. Frohardt 
and Quentin R. Bartz, J. Am. Chem. Soc. 74 599 (1952). 

R. L. Mayer, P. C. Eisman and E. A. Konopka, Experientia 
10 335 (1954). 

728 Phthiomycin, white powder. 

A basic polypeptide resembling viomycin. 
Streptomyces luteochromogenes n. sp. 
Kenji Maeda, Yoshiro Okami, Ryozo Utahara, Hiroko Kosaka 
and Hamao Umezawa, /. Antibiotics (Japan) 6A 183 (1953). 
Yasushi Miyamoto and Kenji Maeda, ibid. 7A 17 (1954). 

729 Streptolin A, Ci^H^iOsN,, or C.4H4,,OiiN-, m.p. 206° (dec), sul- 

fate UW-' -20°. 

Streptolins A and B are similar. They resemble strepto- 
thricin, viomycin, geomycin and roseothricin in their acid 
hydrolysates, which contain L-/;j-lysine, a-D-gulosamine, 
streptolidine, ammonia and carbon dioxide. 



Pfizer Handbook o£ Microbial Metabolites 



352 



Streptomyces spp. 

R. W. Rivett and W. H. Peterson, J. Am. Chem. Soc. 69 
3006 (1947). (Isolation) 

Edward E. Smissman, Robert W. Sharpe, B. F. Aycock, 
Eugene E. van Tamelen and W. H. Peterson, ibid. 75 2029 
(1953). 

Eugene E. van Tamelen and Edward E. Smissman, ibid. 75 
2031 (1953). 

Eugene E. van Tamelen, John R. Dyer, Herbert E. Carter, 
Jack V. Pierce and Edward E. Daniels, ibid. 78 4817 (1956). 

730 Noformicin* (Sulfate), Ci7H3405Nio( 804)2, m.p. (Hydrochlo- 

ride) 265° (dec.). 

Hydrolysis yields glutamic acid, ammonia and two other 
ninhydrin-positive compounds which are not ordinary 
amino acids. 

Nocardia formica 

Dale A. Harris and H. Boyd Woodruff, "Antibiotics Annual 
1953-1954," Medical Encyclopedia, Inc., New York, p. 609. 

731 Streptothricin, CooHj^qOoNs, platelets (Reineckate), m.p. 192-194° 

(Hydrochloride)' [a] d'' -51.3°. 

A basic polypeptide. Hydrolysis yields: 



L-/3-Lysine: 



H2NCH2CH2CH2CHCH2COOH 



NH2 



D-Gulosamine: 



H— C— OH 

I 
H— C— NH2 

I 
H— C— OH 

I 
HO— C— H 

I 
H— C 



Streptolidine: 



CH2OH 

C6H,2N403 



Several structures have been proposed for this moiety. 
See C. Sweeley, Ph.D. Dissertation, Univ. of Illinois, 1955. 
See entry 915 for structure. 



353 



Polypeptides and Related Compounds 



732 



It may be identical with the amino acid known as roseo- 
nine or ge amine. 

OH 

I 
N CH— C— COOH 

II I I 

H2N— C CH2 CH2NH2 

\ / 
N 
H 

Carbon dioxide and ammonia also have been identified 
in hydrolysates. 

Streptomyces lavendulae and other streptomyces species 

Selman A. Waksman and H. Boyd Woodruff, Proc. Soc. Exp. 
Biol. Med. 49 207 (1942). (Isolation) 

Herbert E. Carter, Walter R. Hearn, Edwin M. Lansford, Jr., 
A. C. Page, Jr., Norman P. Salzman, David Shapiro and W. R. 
Taylor, /. Am. Chem. Soc. 74 3704 (1952). (Structure) 

H. E. Carter, R. K. Clark, Jr., Paul Kohn, John W. Rathrock, 
W. R. Taylor, C. A. West, George B. Whitfield and Wilham G. 
Jackson, ibid. 76 566 (1954). 

Koji Nakamishi, Tashito Ito and Yoshimasa Hirata, ibid. 76 
2845 (1954). 

Eugene E. van Tamelen, John R. Dyer, Herbert E. Carter, 
Jack V. Pierce and Edward E. Daniels, ibid. 78 4817 (1956). 

R. Colin, Ph.D. Dissertation, Gottingen, 1957. 

Roseothricins. 

A polypeptide antibiotic complex of the streptothricin 
type. Acid hydrolysis of Roseothricin A yields /? -lysine 
and roseonine (geamine) I in a 1:1 ratio, an isomer of 



OH 

I 
N CH— C— COOH 

HC CH2 CH2NH2 




Pfizer Handbook of Microbial Metabolites 354 

glucosamine, and a substance resistant to further hydroly- 
sis which was assigned structure II. 

Streptomyces roseochromogenes 

Seigo Hosoya, Momoe Soeda, Nobuhiko Komatsu and 
Susumu Imamura, J. Antibiotics (Japan) 4 79 (1951). 

Y. Saburi, ibid. 6B 402 (1953). 

Tashio Goto, Yosimasa Hirata, Seigo Hosoya and Nabukiko 
Komatsu, Bull. Chem. Soc. Japan 30 304, 729 (1957). (Struc- 
ture) 

733 Pleocidin, a hygroscopic white powder. 

A polypeptide resembling streptothricin. 
S. lavendulae or related sp. 

Jesse Charney, Wm. S. Roberts and W. P. Fisher, Antibiotics 
and Chemotherapy 2 307 (1952). 

734, 735 Mycothricins (A and B). 

Basic polypeptides related to streptothricin. Acid hy- 
drolysis yielded ^-lysine, (present in streptothricin, pleo- 
cidin, geomycin and viomycin), roseonine (geamine) 
present in streptothricin, geomycin and pleocidin, and 
serine (present in viomycin). 

Streptomyces lavendulae type 

G. Rangaswami, C. P. SchafFner and S. A. Waksman, Anti- 
biotics and Chemotherapy 6 675 (1956). 

736 Grasseriomycin, pale yellow hydrochloride, m.p. (Reineckate) 

187-190° (dec). Molecular weight 610. 

A polypeptide resembling streptothricin. Negative 
biuret, Millon, FeClg. Positive ninhydrin, Molisch, 
Fehling. 

Streptomyces lavendulae, S. griseolavendus 

Kasububo Ueda, Youichiro Okimoto, Heiichi Sakai and Kei 
Arima, /. Antibiotics (Japan) 8A 91 (1955). 

Yusuke Sumiki, Kinichiro Sakaguchi and Takenori Asai, 
Japanese Patent 6296 (1957). 

737 Actinorubin (C6H14O3N0 or C9H22O4N5) (Hehanthate), reddish 

orange clusters, m.p. 206-214° (dec). 

A basic polypeptide related to streptothricin. Positive 
biuret, reduces KMnOi, Fehlings solution. Negative 
Molisch, Sakaguchi. 

Streptomyces spp. resembling S. erythreus, S. fradii, S. 
albosporeus 

Renate Junowicz-Kocholaty and Walter Kocholaty, /. Biol. 
Chem. 168 757 (1947). 



355 Polypeptides and Related Compounds 

738 Enniatin-B, CooH;{sO,;N._., colorless needles, m.p. 174°, [a],," 

-106.3° (c 0.695 in chloroform). 

CHj 

\ O 

CH3 /H -N^ / 

O CH \ 

/ \ ""■ 

o=c c=o 

CH, \ / 

\ /CH O 

/" N-^r^CH^ CH. 

CHa II CH^ 

CHj 

Fusaria species 
Yield about 0.5 g. per liter. 

PI. A. Plattner and U. Nager, Experientia 3 325 (1947). 
PL A. Plattner, U. Nager and A. Boiler, Helv. Chim. Acta 31 
594 (1948). 

PI. A. Plattner and U. Nager, ibid. 31 665 (1948). 

739 Islanditoxin, C24H31O7N5CI0, colorless, amorphous solid, m.p. 

258°. 

CH2OH O 

o m — <^"— -c 






CHaCH.^^/ CH 



c=o 



^"V ^ -^ C\ CI 



Penicillium islandicum Sopp. 



Pfizer Handbook of Microbial Metabolites 356 

Shingo Marumo and Yusuke Sumiki, /. Agr. Chem. Soc. 
Japan 29 305 (1955). (Isolation) 

Shingo Marumo, Bull. Agr. Chem. Soc. (Japan) 19 258 
(1955). 

Idem., ibid. 23 428 (1959). (Structure) 

740 Enniatin-A ( Lateritiin-I ) , C24H42O6N2, colorless needles, m.p. 
122°, [aW^ -91.9° (c 0.926 in chloroform). 



CH3 



CH,— CH 







\ 


II 
— c — 


CH3 
^ / CH3 


/ 











CH \ 


/ 




\ CH3 


o-c 




c=o 


CH2CH3 \ 




/ 






/ 


^ ^^ 




^CH 


CH3 / 


—c— 


•\ 


CHs 


II 


CH— CH3 



Fusarium orthoceras var. enniatinum, F. scirpi Lamb, 
et Fautr., F. lateritium 

The yield was about 1 g. per liter. 

E. Gaiimann, Stephi Roth, L. Ettlinger, PI. A. Plattner and 
U. Nager, Experientia 3 202 (1947). (Isolation) 

PI. A. Plattner and U. Nager, ibid. 3 325 (1947). 

PI. A. Plattner, U. Nager and A. Boiler, Helv. Chim. Acta 31 
594 (1948). 

PI. A. Plattner and U. Nager, ibid. 31 2192, 2203 (1948). 

A. H. Cook, S. F. Fox and T. H. Farmer, /. Chem. Soc, 1022 
(1949). 



357 Polypeptides and Related Compounds 

741 Enniatin-C, C.^H^.O.jNo, m.p. 123°, [aln" -83° (c 1.162 in chlo- 
roform ) . 

Proposed structure: 

CH3 



CH 

\ 
CH' 



O 



/ \ II CH3 CH3 

CH3^ >H--C--../ -./ 



N. ^CH 



/ 



\ /Ch: \ 



0=9 c==o 



CH3 /CH 

\ /CHr \ / 

CH^ \n^ CH^ CH3 

CHs CH3 II CH 

O \ 

CH3 

Fusaria species 

The yield was about 0.6 g. per liter. 

PI. A. Plattner and U. Nager, Helv. Chim. Acta 31 2203 
(1948). 

742 Eulicin, C24H52O2N8, m.p. (Helianthate) 139°. 

NH 

II 
HoN— C— NH— (CH2)8— CH— CH— (CHsls— NHo 

! I 

OH NH— C— (CHsJsNH- C— NH2 

II II 

O NH 

Streptomyces sp. resembling S. parvus 

An actinomycin and a basic substance also were pro- 
duced. 

Jesse Charney, Roy A. Machlowitz, Frank J. McCarthy, 
Gertrude A. Rutkowski, Alfred A. Tytell and W. P. Fisher, 
"Antibiotics Annual 1955-1956," Medical Encyclopedia, Inc., 
New York, p. 228. (Isolation) 

Robert E. Harman, Edward A. Ham, William A. Bolhofer 
and Norman G. Brink, /. Am. Chem. Soc. 80 5173 (1958). 
(Structure) 



Pfizer Handbook of Microbial Metabolites 



358 



PA 114 Antibiotics. 

743 PA 114A,* Cor.H^iOeNa or C35H42O9N4 (proposed), colorless nee- 

dles, m.p. 200° (dec), [a],,'' -207° (c 0.5 in methanol). 
A neutral substance, green FeCla test. Negative ninhy- 
drin, carbohydrate tests. 

744 PA 114B,t C^oHeaO^oN;, (proposed), colorless crystals, m.p. 265° 

(dec), [alD^-" -59.7° (c 0.5 in methanol). 

A weak acid, red FeCl^ test. Negative ninhydrin, carbo- 
hydrate tests, 2,4-DNPH. 

Streptomyces olivaceus 

Walter D. Celmer and Ben A. Sobin, "Antibiotics Annual 
1955-1956," Medical Encyclopedia, Inc., New York, p. 437. 

745 PA 114B-3, colorless needles, m.p. 207°, [a]i,-'' -37.2° (in 

methanol). 

A polypeptide antibiotic similar to PA-114B. Analysis: 
C 62.77, H6.52, N 12.61. 

A Streptomyces olivaceus strain 

D. C. Hobbs and W. D. Celmer, Federation Proc. 18 246 
(1959). 

746 Streptogramin, approximate formula CocH^^O-N,, m.p. 155°, 

[all, -134°. 

Neutral compound. 

Streptomyces gr amino faciens 

Jesse Charney, W. P. Fisher, Charles Curran, Roy A. 
Machlowitz and Alfred A. Tytell, "Antibiotics Annual 1953- 
1954," Medical Encyclopedia, Inc., New York, p. 171. 

Lateritiin Group 

Several colorless compounds similar to the enniatins 
were isolated from fusaria species in England. One of 
these, lateritiin I, has been shown identical vsdth enniatin 
A. The others are : 



747 

748 
749 
750 



Name 


Suggested formula 


Melting point 


WW 


Lateritiin II 


C26H46O7N0 

C26H,,-,607N2 
C24H«07N2 


125° 

139° 

129° 

86° 


— 92° 




-101° 




-103° 




-83° 







A). 



All these compounds yield D( — )-a-hydroxyiso valeric 
May be identical with staphylomycin M^, E-129A (ostreogrycin 



t See addendum for structure. 



359 Polypeptides and Related Compounds 

acid, C,H,„0,. m.p. 65', [al„''^ -21° (c 1.25 in chloroform), 
and N-methyl-L-valine on acid hydrolysis. 

The enniatins also uniformly contain d( — )-a-hydroxy- 
isovaleric acid, but each contains a characteristic 
N-methylamino acid. (cf. valinomycin, amidomycin). 

A. H. Cook, S. F. Cox, T. H. Farmer and M. S. Lacey, Nature 
160 31 (1947). 

A. H. Cook, S. F. Cox and T. H. Farmer, ibid. 162 61 (1948). 

Idem., J. Chem. Soc, 1022 (1949). 

751 Chlorine-containing Peptide, C2.-,H:{,,0sN-,CL, white needles, m.p. 

251° (dec), [a],/'' -92.9° (in methanol). 

Positive biuret and Pauly reactions, negative Sakaguchi, 
Neubauer-Rhode, ninhydrin, Millon reactions. 

Acid hydrolysis yielded serine (2 to 3 moles), a-amino- 
butyric acid (1 mole), ^-phenyl-/?-aminopropionic acid 
( 1 mole ) and an unidentified substance yielding a posi- 
tive Ehrlich reaction. 

Penicilliinn islandicum Sopp. 

Yoshita Kobayashi, Kenji Uraguchi, Takashi Tatsuno, 
Fuminori Sakai, Michio Tsukioka, Yutaka Sakai, Osamu 
Yonemitsu, Taiko Sato, Masashi Miyake, Mamoru Saito, 
Makoto Enomoto, Toshio Shlkata and Toshitaka Ishlko, Proc. 
Japan Acad. 34 736 (1958). 

752 Pyridomycin, C26-27H32OSN4, colorless needles, m.p. 218-222°. 

Apparently rather closely related to etamycin. Alka- 
line fusion yields : 

OH CH3 CH2CH.3 

, glycine and HOOC— CHCH— COOH 




COOH '^' OH 

Acid hydrolysis yields : 

OH 

, threonine and another degradation product incor- 
•-%^i;:^\ porating picoline and glycine 

■COOH 

Streptomyces pyridomyceticus 

Kenji Maeda, J. Antibiotics (Japan) lOA 94 (1957) and 
earlier papers in the series. 

753 Levomycin, Co7H:^sOtoN,; (proposed), colorless crystals, m.p. 222- 
224°, [a],r' -323° (c 1 in chloroform). 

A polypeptide containing an aromatic group. 
Streptomyces sp. 



Pfizer Handbook of Microbial Metabolites 360 

Herbert E. Carter, Carl P. SchafFner and David Gottlieb, 
Arch. Biochem. and Biophys. 53 282 (1954). 

754 Staphylomycin Mj, CosHagOsNg (probable), m.p. 165-167° (dec), 

[a]D -190° ±2° (c 0.5 in ethanol). 

A neutral compound. Carbonyl group present. Gly- 
cine and proline liberated on acid hydrolysis. Related to 
PA 114A.* 

755 Staphylomycin S, C3g_39H47_4s09N6 (proposed, but see structure 

below), white crystals, m.p. 240-242° [a]i, -28.0° (c 1.0 
in ethanol). 

A weak acid. Threonine, norvaline, a-aminobutryic 
acid, phenylalanine and proline were produced on acid 
hydrolysis. Related to PA 114B (or identical). 

Staphylomycin Mo. This third factor has not been ob- 
tained pure. 

There appears to be a relationship between the staphy- 
lomycin complex and streptogramin, etamycin, etc. 

Streptomyces sp. resembling S. virginiae 

H. Vanderhaeghe, P. Van Dijck, G. Parmentier and P. De 
Somer, Antibiotics and Chemotherapy 7 606 (1957). 

The probable structure of one of the staphylomycins 
recently was reported to be : 











HO 










^r\ 










k } 




- 


/ L-Phenyl- 
NH glycine 


— 


CHa / 
L-Thr CH 

v° 




0^ 


/ 

=^C 

/ L-4-Oxo- 
1 pipecolic 
~CH Acid 

\ 




\ 

NH 


CH2 
0^ 


CH2 




D-a-Amino- 

butyric CH-CH,-CH, 
Acid / 


— cHr 


C L-N-Me 
^ X Phe 
CH 




/ ° 

L-Pro N 






/ ^N 


-Q- 






CH2 / 


11 






/ CH3 

/. A 





^"^^^CHj 






r\ 







♦Identical with PA 114A. 



36i 



Polypeptides and Related Compounds 



H. Vanderhaege, Abstr. Biochem. Symposium, XVIIth 
Internat. Congress Pure and Appl. Chem., Munich 1959. 

H. Vanderhaege and G. Parmentier, Bull. Soc. Chim. beiges 
68 716 (1959). 



756 Phalloidin, C35H46O10NSS + SHoO. 
CHs 



HOCH2 



:=CH— CH— NH 




CHOH 



Amanita phalloides 

From 100 g. of fresh fungus were obtained 10 mg. of 
phalloidin, 8 mg. of x-amanitin, 5 mg. of /?-amanitin and 
about 0.5 mg. of y-amanitin. The amanitins have not 
been characterized thoroughly, but seem to be related to 
phalloidin. 

Theodor Wieland, Angew. Chem. 69 44 (1957). 

Theodor Wieland and Werner Schon, Ann. 593 157 (1955). 

Theodor Wieland and Christoph Dudensing, ibid. 600 156 
(1956). 

757 Phalloin, CssH^gOgNsS, colorless needles, m.p. 250-280° (dec). 
Probable structure: 

CHo CHOH 



CH3 



CH3 



C=CH— CH- 



NH- 



CO- 



CH2 



-CH— NH— CO— CH 



CH2 



^N" 



CH3 



c=o 

I 

NH 

I 
-CH 

I 

c=o 

NH 



X 



-CH 



CO 



CH.. 



CO 

CH— CH3 



CH 



:O^NH 



-NH 



HCOH 

I 
CHs 



Pfizer Handbook of Microbial Metabolites 362 

Amanita phalloides 

Theodor Wieland and Karl Mannes, Angew. Chem. 69 389 
(1957). 

Idem., Ann. 617 152 (1958). 

758 Valinomycin, C;jf;H(;oOi:,N4, colorless platelets, m.p. 190°, [air'" 
+31° (c 1.6 in benzene). 

CH3 

\ II CH-CHa 

^0\-—^ ^NH.^ / 

O /O L-Lactic Acid CH Q 

\ > L-Valine K 

^"\ / \ 

/ CH O CHs 

CH3 7 D-Valine \ / 

W\\ D-a-Hydroxyisovaleric CM \ 

o=c c=o 

^^. \ D-a-Hydroxyisovaleric / 

\ CH Acid NH 

/ \ D-Valine / CH3 

CH3 q /CH / 



C L-Vaiine L-Lactic ,C 

/ \ Acid y V 



CH 



CH3 



NH^ n -CH 



c- 



CH \ \ 



CH3 CHa 



O 



CH 



Streptomyces fulvissimus 

The yield was about 100 mg. per liter. Acid hydrolysis 
gives 2 moles of L( + )-valine, 2 moles of d( — )-vahne, 2 
moles of l( — )-lactic acid and 2 moles of d( — )-a-hydroxy- 
isovaleric acid. (Cf. the enniatin and lateritiin groups, 
and amidomycin.) 

H. Brockmann and G. Schmidt-Kastner, Chem. Ber. 88 57 
(1955). 

Hans Brockmann and Hermann Geeren, Ann. 603 213 
(1957). 

759 Viscosin, C;{,.H,j,;0]„N,;, amorphous white powder, m.p. 269° 
(dec), [a],r" -162°. 



363 Polypeptides and Related Compounds 

:H3(CH )6— CH— CH2— CO— NH CH CO— NH— CH. CO NH CH CO— 

I I " I 

OH CH, CHo 

I 1 

CH OH 

/ \ 

CH3 CH:, 

NH— CH— CO— NH— CH— CO— NH— CH— COOH 

I I I 

CH CHOH CH2 

/ \ I I 

CH3 CHa CH3 CH 

/ \ 
CH3 CH3 

Pseudomonas viscosa 

Mitsuyuki Kochi, Vincent Groupe, Leonora H. Pugh and 
David Weiss, Bad. Proc, 29 (1951). 

Takashi Ohno, Shigeru Tajinia and Katsuyuki Toki, /. Agr. 
Chem. Soc. Japan 27 665 (1953). 

Doki and Ohno (unpublished). Total structure determina- 
tion. Reported by S. Otani in a lecture on polypeptide anti- 
biotics in 1957. 

Takashi Ohno, Shigeru Tajima and Katsuyuki Toki, /. Agr. 
Chem. Soc. Japan 27 665 (1953). 

760 Bottromycin (B-Mycin), C3sH57_r,i07_.sN7S, white amorphous ma- 
terial, [a],;--' -14.2° (c 0.5 in 96%, ethanol). 

Bottromycin is a weakly basic polypeptide. Acid hy- 
drolysis yields six ninhydrin-positive compounds. Two of 
these are glycine and valine. Two others are new amino 
acids : 



a-Amino-/3-phenylbutyric Acid 



CH CH— COOH 



CH3 NH2 

and 
/3-(2-Thiazole)-/3-alanine 

CH— CHo— COOH 

I 
NH2 

Streptomyces bottropensis 



Pfizer Handbook of Microbial Metabolites 364 

J. M. Waisvisz, M. G. van der Hoeven, J. van Peppen and 
W. C. M. Zwennis, J. Am. Chem. Soc. 79 4520 (1957). (Iso- 
lation ) 

J. M. Waisvisz, M. G. van der Hoeven, J. F. Holscher and 
B. te Nijenhuis, ibid. 79 4522 (1957). 

J. M. Waisvisz, M. G. van der Hoeven and B. te Nijenhuis, 
ibid. 79 4524 (1957). (Structure) 

Micrococcins. 

761 Micrococcin, white needles, m.p. 222-228° (dec.), [aln^^ 116° 

±1° (c 5.0 in 90% ethanol), molecular weight >2000. 

A Micrococcus sp. 

T. L. Su, Brit. J. Exptl. Path. 29 473 (1948). 

N. G. Heatley and Hazel M. Doery, Biochem. J. 50 247 
(1951). 

762 Micrococcin-P, white crystals, yellowing in light, m.p. 252° (dec. 

from 232°), [ale"' +63.7° (c 1.19 in 90% ethanol), mo- 
lecular weight '-'2200. 

Two fragments have been identified as : 



HOOC HOOC 

C— CH2CH3 CH— CH 



CH3 



O NH2 CH3 

2-Propionylthiazole-4- 2-(l -Amino-2-methylpropyl) 

carboxylic Acid thiazole-4-carboxylic Acid 

Acid-catalyzed esterification gave a dimethyl ester, 
C24Ho365Nr,S4 and a base CifiHictO^N^S-^. Also threonine, 
ammonia and propionic acid were isolated. 

This antibiotic seems to be similar to or identical with 
the earlier one, but is distinguished by the letter P until 
identity is proved. 

Bacillus pumilis 

A. T. Fuller, Nature 175 722 (1955). (Isolation) 

E. P. Abraham, N. G. Headey, P. Brookes, A. T. Fuller and 
James Walker, ibid. 178 44 (1956). 

P. Brookes, A. T. Fuller and James Walker, J. Chem. Soc, 
689 (1957). 



3^5 Polypeptides and Related Compounds 

763 Esperin, CsgH^-OnN-, colorless crystals, m.p. 238° (dec ) [al,^'^ 

-24° (c 0.66 in methanol). 

CH,CH,COOH 

CHslCH.lgCHCH.CONHCHCONHCHCONHCHCONHCHCONHCHCOOH 

I I I 

CH CH, 

O OC— CH., / \ CH- I 

CHa CH3 I CH 

CH /\ 

/ \ CH3 CH3 

CH3 CH3 

Bacillus mesentericus 

Hiroshi Ogawa and Teiichiro Ito, ]. Agr. Chem. Soc. Japan 
24 191 (1950). (Isolation) 

Idem., ibid. 26 432 (1952). 

Idem., Bull. Agr. Chem. Soc. (Japan) 23 536 (1959). 
(Structure) 

764 Actinochrysin, C4oHr.-0„N-, a brick red pigment. 

Similar to but distinct from actinomycins. A weak 
base with two acid groups. Molecular weight 811. Solu- 
ble in acetone. 

Streptomyces chrysomallus 

Hans Brockmann and Arnold Bohne, German Patent 912,- 
010 (1954). (Chem. Abstr. 52 12334g) 

765 Grisein, C4i,H(;i02„Ni„SFe, red, amorphous powder. 

Isolated from acid hydrolysate: 

O H 

I ll + Glutamic Acid 



y^.^J^ + '^n unidentified amino acid 
CH, II 

3-Methyluracil 

The iron is Fe'" and can be removed and readded to the 
complex. 

Streptomyces griseus 

The Russian antibiotic, albomycin, produced by Strepto- 
myces subtropicus seems to be similar to or identical with 
grisein. 



Pfizer Handbook of Microbial Metabolites 366 

Donald M. Reynolds, Albert Schatz and Selman A. Waks- 
man, Proc. Soc. Exp. Biol. 64 50 (1947). (Isolation) 

Donald M. Reynolds and Selman A. Waksman, /. Bad. 
55 739 (1948). 

Frederick A. Kuehl, Jr., Mary Neale Bishop, Louis Chaiet 
and Karl Folkers, J. Am. Chem. Soc. 73 1770 (1951). 

766 Albomycin (Sulfate), red amorphous powder, molecular weight 

>1300. 

Partial Constitution: 

Albomycin is a basic, cyclic polypeptide containing iron 
(^4% by weight). Iron can be removed with acetone 
(color loss) and restored with FeCl^. Hydrolysis yields: 
ornithine, serine, glutamic acid, alanine, glycine, proline 
and one unidentified amino acid. 

Streptomyces siibtropicus n. sp. 

Albomycin may be identical with grisein, produced by 
Streptomyces griseus. 

M. G. Brazhnikova, N. N. Lomakina and L. I. Murayeva, 
Dokladij Akad. Nauk S.S.S.R. 99 827 (1954). 

G. F. Gause, Brit. Med. J. 2 1177 (1955). 

Yu. O. Sazykin, Mikrobiologiya 24 75 (1955). 

767 Amidomycin, C4i|H,.sOi2N4, colorless needles, m.p. 192°, [a]i)^*' 

+ 19.2° (c 1.2 in ethanol). 

CHj CH3 
\ / CH3 CH3 

CH O \ / 

\ It CH 

^CH C — -Q / 

CH3 O. /NH CH o 



CH 



D-Hiv 



/ ^CH D-Hiv NH CH3 

^ i / 

o 



o=c 

CH3 \ 

rw— CH D-Val 

/ \ 

CH3 NH 

\ 

CH 

/ \ 

CH3 CH:, 





\ 


/ 
-CH 




D-Val CH' 

\ 


\ 
CHj 




c= 


=0 




/ 













D-Hiv CH /^"^ 
_/ CH 


D-Val 


y^r. 


\ 


^NH 


1 


CH3 


— CH 






\ 






CH 






/ \ 






CH3 CHj 







Streptomyces species (PRL 1642) 

Amidomycin contains 4 moles each of d( — ) valine and 
d( — ) a-hydroxyisovaleric acid. (Cf. Valinomycin, lateri- 
tiins, enniatins.) 

L. C. Vining and W. A. Taber, Can. J. Chem. 35 1109 
(1957). 



3^7 Polypeptides and Related Compounds 

768 Toxin of Helminthosporium victoriae. 

This toxin consists of two loosely connected moieties. 
The first is a tricyclic secondary amine called victoxinine, 
and the second a pentapeptide. The intact toxin shows a 
negative ninhydrin test, and a molecular weight of 800 
was assumed. 

Victoxinine, C,-Ho,,ON (Hydrochloride), colorless nee- 
dles, m.p. 172°, [a],r' -78° (c 3.2 in 95% alcohol). 
Negative U.V. 

Pentapeptide : 

On acid hydrolysis yielded: 
Aspartic acid, glutamic acid, glycine, valine and one of 
the leucines. 

Helminthosporiinn victoriae 

Ross B. Pringle and Armin C. Braun, Nature 181 1205 
(1958). 

769 Telomycin, cream colored amorphous solid. 

A polypeptide antibiotic, containing glycine, alanine, 
threonine and aspartic acid. Molecular weight about 
1000. Contains no sulfur. Negative Fehling, ninhydrin, 
biuret. Similar to etamycin. 

Streptomyces sp. 

M. Misiek, O. B. Fardig, A. Gourevitch, D. L. Johnson, I. R. 
Hooper and J. Lein, "Antibiotics Annual 1957-1958," Medical 
Encyclopedia, Inc., New York, p. 852. 

770 Etamycin (Viridogrisein), C44H6oOi,.N,s, white crystals, hydro- 

chloride m.p. 163-170° (dec), [a]ir' conflicting reports. 

HO, 
O 

\ ^^C -^"^CH^ / 
CH3 CH ^CH 



/N Sore \^ 

o 



P NH CH, 

/ \ ^ ^ 

CHj— CH L-Ala D-Leu CM— CH2 ^H 

I 1 ^"' 

NH C=o 

\ I 

" \ OH-Pro / CH, 

CH 3,N-DIMe CH \ 

C\ Ch' X^ Leu Sore ^ CH"^"^ 

CH CH, / C- CH ^^ Au 

/ ^"^ CH3 // ^"^ \ O OH 

CHj o CH, 



Pfizer Handbook of Microbial Metabolites 



368 



Streptomyces sp. resembling S. lavendulae 

Cf. Pyridomycin staphylomycin, osteogrycin, PA-114, 
mikamycin, streptogramin, telomycin, echinomycin. 

This class of polypeptides appears to be related bi- 
ogenically to the actinomycins. 

B. Heinemann, A. Gourevitch, J. Lein, D. L. Johnson, M. A. 
Kaplan, D. Vanas and I. R. Hooper, "Antibiotics Annual 1954— 
1955," Medical Encyclopedia, Inc., New York, p. 728. 

Quentin R. Bartz, Jean Standiford, James D. Mold, Doris W. 
Johannessen, Albert Ryder, Andrew Maretzki and Theodore H. 
Haskell, ibid., p. 777. 

Theodore H. Haskell, Andrew Maretzki and Quentin R. 
Bartz, ibid., p. 784. 

John C. Sheehan, Hans Georg Zachau and WiUiam B. Law- 
son, /. Am. Chem. Soc. 79 3933 (1957). (Structure) 



771 Colistin, C4gH850ioNi3. 



CH3 



CH3 



\ 

I 

/ 



co- 



NHo 

CH2 
I 
CH2 

I 
-CH- 



-NH.^ 



CHCH2 NH' 



CO 



/ 



NH 
HaNCHjCrij— CH 

I 

CO 

\ 

NH 



i-Diamino- ^CO CH2CH2NH2 

butyric Acid \^ / 

CH 
D-Leucine L-Diamino- \ 

butyric Acid \li 

\ 

CO 

l-Diaminobutyric \ 

Acid CH — CH0CH2NH2 

NH 

/ 



aminobutyric 
Acid 



CH3 



L-Leucine 



\ 



CH 



CHCHf 



CO. 



L-Diaminobutyric 
Acid 



L-Threonine CO 

/ 

CH 
NH CH 



OH 



-NH 



CO' 



\ 



CH3 



CH3 



-CH- 

] 
CHo 

I 
CH2 

I 

NH— CO(CH2)4CHCH2CH3 
I 
CHs 

Yasuo Koyama, Akio Kurosasa, Atsushi Tsuchiya and Kin- 
suke Takakuta, J. Antibiotics (Japan) 3 457 (1950). 



3^9 Polypep t ides a nd Related Compounds 

Taiichi Ito, Sadao Miyamura, Seihachiro Niwayama, Masa- 
nobu Oishi, Nobuhiro Igarashi, Hiromichi Hoshino and Shozo 
Muto, ibid. 7B 147 (1954). 

Yasuo Kayama, Japanese Patent 1546 (1952). 

Takeshi Oda, Mitsuhiro Kinoshita, Osamu Yamanaka and 
Fumio Ueda, /. Pharm. Soc. Japan 74 1234 (1954). 

Takeshi Oda and Fumio Ueda, ibid. 74 1246 (1954). 

772 Mycobactin, C^-H^.^Oi^N-, microcrystalline white powder with 
pale green fluorescence, m.p. 165-166.5°, [a],,'' -19° (c 
4.9 in chloroform). 

Mycobactin is a weak acid believed to have one of the 
following structures: 




O— CH2 

/ 

ru C COOR OH 

^"^ \ II 

N— CH— CO— NH— CH— (CH2)4— N— CO— CH=CH(CH2)uCH3 

trans 

or 




CH3 



C OH COOR 

\ 

N— CH— CO— N— (CH.)4— CH— NH— CO— CH=CH(CH2),4CH3 

trans 

CH2— CH,— CH2 

I / 

R = CH3CH2CHCHCONH— CH 

I \ 
CH3 CO N CH2 

OH 

Mycobacterium phlei 

The yield was about 67 g. from 41 kg. of moist cells. 
G. A. Snow, J. Chem. Soc, 4080 (1954) and earlier papers 
in the series. 

73 Geomycin (C,jHio02N2)8-i,„ HeHanthate red platelets, m.p. 205- 
215° (dec). Hydrochloride [a],,-" +16°. 

A basic polypeptide. Acid hydrolysis yields: geamine, 
/3-lysine, and an amino sugar, plus small amounts of 



Pfizer Handbook of Microbial Metabolites 



370 



aspartic acid, glutamic acid, serine, threonine, glycine 
and alanine. 

The structural evidence has been well summarized and 
a partial structure postulated by R. Colin, Ph.D. Disserta- 
tion, Gottingen, 1957. The partial structure is: 

NH NH2 

-C— NH— R^ 

O 

HOOC— C CH N 

I I II 

CH2 CH2 C 

I \m/ 

NHo ^^ 





"O 






NH 


C— CH2- 


-CH— (CH2)3- 


-NH 


(CHo)3 


_ 


NH2 


_ 


CH— NH2 

1 






1 
CH2 






c=o 






NH2 









NH2 



Streptomyces xanthophaeus , n.sp. 

Hans Brockmann and Burchard Franck, Naturwissenschaf- 
ten 41 451 (1954). 

Hans Brockmann and Hans Musso, Chem. Ber. 87 1779 
(1954). 

Idem., ibid. 88 648 (1955). 

774 Lavendulin (Helianthate), C49H63O18N13S (proposed), orange 

crystals, m.p. 212-220° (dec). 

A basic polypeptide. Positive FeCIs, Fehhng, biuret, 
KMn04. Negative Molisch, Sakaguchi. 

Streptomyces sp. similar to S. lavendulae 

Albert Kelner and Harry E. Morton, J. Bad. 53 695 (1947). 
(Isolation) 

Harry E. Morton, Proc. Soc. Exp. Biol. Med. 64 327 (1947). 

775 Echinomycin (X-948),* C-,„H„„Oi2Ni^,S2, granular, nearly color- 

less hygroscopic powder, m.p. 217°, [a]i> —310° (c 0.86 
in chloroform). 



CH-CO NH— CH— CO— N - C— CO -N— CH— CO- O CH. 

/ \ 
S CH, 

I I 

CH: S 

CH,— O— CO— CH— N— CO-C-N CO— CH NH-CO— CH 

II II 

CH CH, CH, CH, 

/ \_ I 



CH, 



CH, 



CO 



* Antibiotic X-1008 (unclassified) resembles echinomycin. 



371 Polypep tides and Related Compounds 

Streptornyces echinatus n. sp. 

R. Corbaz. L. Ettlinger, E. Gaumann, W. Keller-Schierlein, 
F. Kradolfer, L. Neipp, V. Prelog, P. Reusser and H. Zahner' 
Helv. Chim. Acta 40 199 (1957). (Isolation) 

W. Keller-Schierlein, M. Lj. Mikhailovich and V Prelog 
iWd. 42 305 (1959). (Structure) 

Circulins, C-,3Hi„„Oi3Ni(5 (Sulfate), amorphous solid, m.p. 226- 
228° (dec), [«]„" -61.6°. 

776 Circulin A: 



CHs O 

I II 

CH3CH2CHCH2CH2CH2CH2— C— NH 

NH2 ^"- NHo* 

\ ru O / 

CH ^CH O 

% / \ 

CH. C NH 



CH3 



CH3 



CH3 / 

NH 



CH' 



CH L-Isoleu t-Thre CH' 



\ OH 

c==o 



0=C NH 

-„.^CH D-Leu l-Dia CH 

,CH2 \^ / 



CH2 

^CHo 



/ NH C^ --KJU 

CH^ L-Thre CH 

CH2 N"^^ ^-Dia ^C^ \ „ ^^ 

CH2 o' J.^ O CH3 



H2N 



/ 



CH2 
CH2 

I 

NHo 



Bacillus circulans 

Hydrolysis yields 6 moles of L-a, y-diaminobutyric acid, 
2 moles of L-threonine, 1 mole of D-leucine, 1 mole of 
L-isoleucine and 1 mole of ( + )6-methyloctanoic acid. 

'7 Circulin B has essentially the same structure, but the 6-methyl- 
j octanoic acid moiety is attached at the starred amino 



Pfizer Handbook of Microbial Metabolites 372 

group. There may be other similar compounds in the 
complex also. 

F. J. Murray, P. A. Tetrault, O. W. Kaufman, H. Koffler, 
D. H. Peterson and D. R. Colingsworth, J. Bad. 57 305 
(1949). 

D. H. Peterson and L. M. Reineke, J. Biol. Chem. 181 95 
(1949). 

Tashio Kobayashi, J. E. Grady, J. L. Parsons, Henry Koffler 
and P. A. Tetrault, Abstr. 133rd Meeting Am. Cheyn. Soc, 25C 
(1958). 

H. Koffler and T. Kobayashi, Abstr. 4th Intern. Congr. 
Biochem., 9 (1958). 

Henry Koffler, Science 130 1419 (1959). 

778 Fungisporin, Cr.gHyoOgNg, colorless crystals, m.p. 355-360° 
(dec.) (subl. from 250°), molecular weight 980. 
Proposed structure: 

CHs CH3 

CH O 




O 1 // CH3 

\ X D-Valine \ / \ 

CH L-Phenyl- CH CH3 

// alanine L-Valine \ q 

NH C^ 

=C NH 

/ '^— CH2— CH D-Phenyl- D-Phenyl- CH— CH.— '^^^ 

1 alanine alanine I 

NH C^^ 

\ / ° 

^C NH 

O^ \ L-Valine L-Phenyl- / 

CH3 CH alanine CH 



CH 



\ / \ D-Valine _/^ \ 

CH NH. Xf Yh 

^C-^CH—NH-^X CH. 

// I o 

O CH 

/ \ 
CH3 CH3 

Penicillium and Aspergillus spp. 

This polypeptide was obtained by destructive distilla- 
tion of spores, when it separated by sublimation. 



373 Polype ptides and Related Compounds 

U. Sumiki and K. Miyao, /. Agr. Chem. Soc. Japan 26 27 
(1952). 

Idem., Bull. Agr. Chem. Soc. (Japan) 19 86 (1955). 
Kohei Miyao, ibid. 24 23 (1960). 

779 Polypeptin (formerly called circulin, but not identical with the 
polypeptide now known as circuHn), C^gHooOiaN,.., color- 
less crystals, m.p. 176°, [a],,-" (Sulfate) -93.3° (c 3.0 in 
70% isopropyl alcohol). 

A basic polypeptide, containing: three a,y-diaminobu- 
tyric acids, one L-threonine, one D-vaHne, one L-isoleucine, 
two L-leucines and one D-phenylalanine. 

Bacillus krzemieniewski, a B. circulans mucoid variant 

Stacey F. Howell, /. Biol. Chem. 186 863 (1950). 

Werner Hausmann and Lyman C. Craig, ibid. 198 405 
(1952). 

Polymyxins: 

'80 Polymyxin A (Aerosporin) 

'81 Polymyxin Bi 

'82 Polymyxin B, 

'83 Polymyxin C 

'84 Polymyxin D 

'85 Polymyxin E 

A complex of related polypeptides produced by Bacillus 
polymyxa. Initially five components, A, B, C, D and E 
were separated. Then B was resolved into two compo- 
nents Bi and B^, differing only in the fatty acid moiety. 
All polymyxins contain L-a, y-diaminobutyric acid and 
L-threonine. All but B^ apparently contain d-6-methyloc- 
tanoic acid, and it contains a C-8 acid instead. Poly- 
myxin A has been reported to contain D-leucine but no 
phenylalanine. It is also known as aerosporin because it 
is produced by Bacillus aerosporus. Polymyxin C con- 
tains phenylalanine but no leucine. Polymyxin D con- 
tains leucine but no phenylalanine, and it also has been 
reported to contain D-serine. Polymyxin E has the same 
quahtative composition as A, but is distinct. 

Two alternative structures have been suggested for 
polymyxin Bj as the result of degradative studies. These 
structures are shown here, the amino acids being abbrevi- 
ated in the following manner: 



Pfizer Handbook of Microbial Metabolites 



374 



HjN- 



Dia = a,y-Diaminobutyric Acid 
Thr = Threonine 
Phe = Phenylalanine 
Leu = Leucine 

NH2 NH, 

\ I 

CH, I I 

).„ O O CH— OH O CHi 

Jh J-,H^ /CH.-CH.MH-C-CH-NH-C-CH 
CH X ''°'° l-Dia \^ I 

/\/ \ c=o 

CH3 CH L-Thr NH I 

/ \ (CH2), 

NH L-Dia CH^^"^^"^N"2 



/ 

o=c 



,CHr 



CH— CH3 

1 I 

\ r 1"" 

CHo-'S" i-Dia NH ^Hj 

\ y 6-Methy|. 

NH o-Phe CH octanoic 

\ / ^CH ^'''^ 



CHo O 



CH2 



/ ■ ^CH 



NH: 



CH3 CHj 



NHo 



Or CH2 CH3 OH 

1 \ / 

CH, OH O CH2 O CH 

\ / " II I II I 

\C O CH,— NH— C— CH— NH— C— CH 

\ II / I 

CH C — .^^ CH; ^.Dio L.Thr NH 

a NH "^CH I „ 

X^/ L-Thr ,.Dia \ .O C=0 

^"'--rw / \ CH-(CHo)oNH2 

9" NH NH 

NH i-Dia CH— CHj— CH.NH. C=0 



= C C^ 

\ L-Dio /"° 



I 

(CHJ^ 



CH 

^CH ^(l / \ 

/CH. \ ^ o-Phe / ^H. CH3 

CH2 X. l-Leu CH ^.j^ 

H2N C-^ , _C^ \ 



^^— CH-NH--^^ "\^ 

CH 

/ \ 
CH3 CH3 



octanoic 
Acid 



375 



Polypeptides and Related Compounds 



Commercial polymyxin is essentially polymyxin B sul- 
fate, a white powder, m.p. 228-232° (dec.), Whr' -45° 
(c 0.1). The empirical formula of the free base is 
CseH.tc.gsOjsNie. 

G. C. Ainsworth, A. M. Brown and G. Brownlee, Nature 160 
263 (1947). (Isolation) 

George Brownlee, Ann. N. Y. Acad. Sci. 51 875 (1949). 
(Polymyxin A) 

P. H. Bell, J. F. Bone, J. P. English, C. E. Fellows, K. S. 
Howard, M. M. Rogers, R. G. Shepherd and R. Winterbottom, 
ibid. 51 897 (1949). (Degradations, identification of amino 
acids) 

Tudor S. G. Jones, ibid. 51 909 (1949). (Separations, deg- 
radations, identification of amino acids) 

J. R. Catch, Tudor S. G. Jones and S. Wilkinson, ibid. 51 
917 (1949). 

P. P. Regna, I. A. Solomons, B. K. Forscher and A. E. 
Timreck, /. Clin. Invest. 28 1022 (1949). (Purification of B) 

Werner Hausmann and Lyman C. Craig, J. Am. Chem. Soc. 
76 4892 (1954). (Resolution of B into two parts) 

Werner Hausmann, ibid. 78 3663 (1956). (Proposal of de- 
tailed cyclic structures) 

Gerard Biserte and Michel Dautrevaux, Bull. soc. chim. biol. 
39 795 (1957). (Structure) 

Gramicidins. 

A mixture of polypeptides produced by Bacillus brevis 
and originally called tyrothricin was separated into two 
groups, the tyrocidines (about 80 "^r ) and the gramicidins 
(about 20%). Each of these groups has been fraction- 
ated further into pure polypeptides. 

The original gramicidin consisted of a mixture of three 
closely related neutral polypeptides. It was assigned an 
average empirical formula of C]4SjH2io02(iN3o, colorless 
platelets, m.p. 228-231°, [aW +3°. 



Fraction A 


Fraction B 


Fraction C 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


+ 




+ 


+ 



D-Leucine 

L-Trypotophon. 

l-Alanine 

DL- Valine 

Glycine 

Phenylalanine. 
Tyrosine 



Pfizer Handbook of Microbial Metabolites 



376 



RoUin D. Hotchkiss and Rene J. Dubos, /. Biol. Chem. 132 
791 (1940). 

Idem., ibid. 141 155 (1941). (Isolation) 

Max Tishler, J. L. Stokes, N. R. Trenner and John B. Conn, 
ibid. 141 197 (1941). 

Rollin D. Hotchkiss, Advances in Enzymol. 4 153 (1944). 



786 Gramicidin Jo, C35H56O6N8. 




NH2 

1 




1 
CH2 

1 




1 
CH2 

1 




1 
CH2 

1 





1 
^r^ D-Orn 
CH2 CH 


^cC^ 


NH 

\ 

L-Val CH 



.CH 



\ / 



CHa 



CH3 



"N i-Pro 



o=c 



\ 



CH D-Phe 



NH 

/ 

L-Orn CH 



CH2 



CH2 



NH 



/ 



D-Leu 
-CH^ 

CH2 

I 
CH 



-NH 



\ 



XH2 



CH2 



CH3 



CH3 



'NH2 



Bacillus brevis 

Shokei Otani, H. Nagano and Y. Saito, Osaka Shiritsu 
Daigaku Igaku Zasshi 7 640-650 (1958). (Chem. Abstr. 
12403g) 



377 Polypeptides and Related Compounds 

787 Gramicidin J,, C44H6.r,0-N{,. 



NH2 

I 
CH2 

I 
CH2 

I 

CH2 o 

_— -CH2 S- ^ NH CH 



CH CH 

/ \ 



CH3 



/^y_CH2 y 



■CH2 



/ ^NH Qr^ \ 

CH 



CH 

6 




-CH2 



NH C ^NH2 

\ / O 

>\ D-Phe y 

-^ ^CH ^CH^ 



/ 
CH 

/ \ 
CH3 CH3 



Bacillus brevis 

Shokei Otani and Yoshitaka Saito, Proc. Japan. Acad. 30 
991 (1954). 

Idem., Congr. intern, biochim.. Resumes Communs., 3e 
Congr., Brussels, 88 (1955). 



Pfizer Handbook of Microbial Metabolites 378 



788 Gramicidin S (Gramicidin C), CeoHgoOioNio, colorless needles, 
m.p. 277° (dec), [2;]..'' -289° ±10° (c 0.43 in 70% etha- 
nol). 





NH2 






CH2 


CHs CHs 


CH3 CHs 


CHo 


CH 


\/ 


1 


1 


CH 


CH2 


CH2 



CH 



CH2 



CO— NHCHCO— NHCHCO— NHCHCO— NHCHCO— N CHo 

I (l) (l) (l) (d) 
CH,— CH 

[l) (l) CH— CH'2 

(d) (l) (l) (l) 1 

CH2— N COCHNH— COCHNH— COCHNH— COCHNH— CO 

I I I 1 

CH. CHo CH2 CH 

I I I / \ 

CH CH, CHs CHs 

/ \ 1 

CHs CHs CH2 

I 
NH2 

Bacillus brevis var. Gause-Brazhnikova 

G. F. Cause and M. G. Brazhnikova, Am. Rev. Soviet Med. 
2 134 (1944). 

R. L. M. Synge, Biochem. }. 39 363 (1945). (Character- 
istics) 

F. Sanger, ibid. 40 261 (1946). 

R. Consden, A. H. Gordon, A. J. P. Martin and R. L. M. 
Synge, ibid. 40 xciii (1946). 

Idem., ibid. 41 596 (1947). 

Alan R. Battersby and Lyman C. Craig, /. Am. Chem. Soc. 
73 1887 (1951). 

R. Schwyzer and P. Sieber, Helv. Chim. Acta 40 624 (1957). 
(Synthesis) 

789 Gramicidin D (Gramicidin Dubos), colorless crystals, m.p. 229° 
(dec). 

A crystalline component of tyrothricin. A cyclic poly- 
peptide composed of 4 moles of D-Leucine, 4 moles of l- 
tryptophan, 2 moles of o-Valine, 2 moles of L-Valine, 2 
moles of L-alanine, 1 mole of glycine and 1 mole of 
ethanolamine. 

Bacillus brevis 

Rene J. Dubos and Rollin D. Hotchkiss, J. Exptl. Med. 73 
629 (1941). (Isolation) 

A. H. Gordon, A. T. P. Martin and R. L. M. Synge, Biochem. 
J. 37 86 (1943). 



379 Polypeptides and Related Compounds 

Rollin D. Hotchkiss, Advmices in Enzymol. 4 153 (1944). 

R. L. M. Synge, Biochem. J. 39 355 (1945). 

T. S. Work, The relation of optical form to biological 
activity in the amino acid series, Biochem. Soc. Symposia 1 
61 (1948). 

790 Racemomvcin B, C,;oHio>,0;,.,N..„. white, hygroscopic powder, m.p. 

150°, [2W -34° (c 6.5 in water). 

A basic antibiotic resembhng streptothricln. Acid 
hydrolysis gives a reducing sugar and carbon dioxide, 
^-lysine and roseonine in the ratio 2:3:2. Racemomycin 
B occurs in a complex with two (apparently similar) sub- 
stances, racemomycins A and C. 

Streptomyces racemochromogemis n. sp. 

Hyozo Taniyama and Shoji Takemura, J. Pharm. Soc. Ja- 
pan 77 1210 (1957). 

Idem., ibid. 78 742 (1958). 

791 Tyrocidine A, C66Hji60i3Ni3, colorless needles or rods, m.p. 240- 
242° (dec), [a]v-'' -111°. A component of the tyrothri- 



cin complex. 



NH, 



CH2 

I 

I /"^ 

CH3 CH, o CH 

I O I // / \ 

CH3-CH \^NH-CH-^C^ / CH3 

CH i-Orn ^CH ^ 

X L-Val L-Leo \ /^ 



CH L-Tyr D-Phe CH"^ " 

NH C=0 

I 1 

0-=C N — -CH2 

^CH i-Glu i-Pro CH._ ^ 

CH=-^"- \ / ^"= 

OC-^ NH C^ 

\ / % 

O ^CH L-Asp ^.p,^ CH 

r/ NH_ .C^ \ 

CH. --C^CH-NH-^ \\ CH. 

OC I \ ° 

/ ° CH2 

H2N I \ J 



CH, 



Pfizer Handbook of Microbial Metabolites 380 

Bacillus brevis 

Rollin D. Hotchkiss and Rene J. Dubos, /. Biol. Chem. 132 
791 (1940). (Isolation) 

R. L. M. Synge and A. Tiselius, Acta Chem. Scand. 1 749 
(1947). 

R. L. M. Synge, Quart. Rev. 3 245 (1949). (Review of 
work to that date) 

Alan R. Battersby and Lyman C. Craig, /. Am. Chem,. Soc. 
74 4019, 4023 (1952). (Separation) 

Alejandro Paladini and Lyman C. Craig, ibid. 76 688 
(1954). (Structure) 

792 Tyrocidine B, C68H88O13N14. 

A component of the tyrothricin complex. 




H2NOC 



381 Polypeptides and Related Compounds 

Bacillus brevis 

T. P. King and L. C. Craig. /. Am. Chcm. Soc. 77 6627 
(1955). (Final structure) 

Actinomycins. 

The nomenclature of the actinomycins is confused be- 
cause they occur in difficulty separable complex mixtures, 
several different research groups have investigated them, 
and, even when pure, one substance cannot be compared 
with another by techniques as simple as a mixed melting 
point. This problem has been discussed by Brockmann 
in a review of the actinomycins. 

L. Zechmeister (editor), "Fortschritte der Chemie organis- 
cher NaturstofFe" XVIII, Hans Brockmann, The actinomycins. 
Springer Verlag, Vienna, 1960. 

At first actinomycins A, B and C were isolated, but later 
these were found to be mixtures. As such complexes 
were resolved by paper chromatography, Arabic numeral 
subscripts were attached to the capital Roman letter in 
order of appearance on the developed chromatogram, the 
origin on the paper being zero (e.g., Ci, Co, C3). When 
some of the separated actinomycins were resolved even 
further, a further subdivision in nomenclature was re- 
quired; so a lower case Roman letter was attached to give, 
^■9-, Coj, which appeared between Co and C3. When the 
Xo complex at the origin was resolved, a slightly different 
system was used, Greek letters being attached to the 
Arabic numeral subscript, e.g., Xo^ was less polar than X,,;,. 

Few series are complete because often names have been 
eliminated due to duplication, further resolution, etc. 
Thus, a complex designated I was resolved into Ii and L, 
but these later were shown to be the same as Cj and C2 
and the I names eliminated. 

Still this method of nomenclature does have a ration- 
ale, although it may not be readily apparent, and it is used 
in Germany and in Switzerland. 

Other groups continue to refer to various complexes as 
A, B or D types. These consist essentially of various ratios 
of actinomycin Xo and its reduction product, actinomycin 
Ci, actinomycin D being nearly pure Cj. 

The E and F series arose when it was discovered that 
addition of certain amino acids to the medium in large 
amounts caused displacement of certain other amino 
acids in the peptide side-chains, thus creating new "bio- 
synthetic" actinomycins. 



Pfizer Handbook of Microbial Metabolites 



382 



Beyond historical interest there seems to be little point 
in attempting to standardize the nomenclature of actino- 
mycin mixtures. Waksman has proposed that a Roman 
numeral be assigned to each pure actinomycin, and John- 
son's group has taken up this practice, actinomycins II 
and III being distinct from those characterized elsewhere, 
while IV is identical with Cj or D, etc. Brockmann views 
this as one more contribution to the confusion of the liter- 
ature and, claiming the right of discoverer of many of the 
actinomycins, has made the suggestion that no nomen- 
clature system will relieve the confusion unless it makes 
apparent the amino acid sequences of the side-chains. 

Although this does not solve the problem of trivial 
nomenclature, Brockmann uses a shorthand method of 
demonstrating the structures of the actinomycins in 

which a symbol < represents the actinocinin moiety, 

the branches at the right symbolizing the amino and 
quinonoid carbonyl groups. The abbreviated amino 
acid names are then attached in proper sequence. In 
most of the asymmetric actinomycins the chains in which 
the differing amino acids occur have not yet been speci- 
fied, and this is indicated by an E -symbol, indicating pos- 
sible reversal of position. 

The structure of actinomycin C3 (which has been 
synthesized) is given somewhat more fully to show struc- 
tural details. The custom of arrangement by empirical 
formula is ignored here to permit grouping by related 
structures. 

The mitomycins (unclassified) may be actinomycins. 

There is an apparent striking biogenetic similarity 
among the etamycin, staphylomycin, etc. group of poly- 
peptides on the one hand and the actinomycins on the 
other. 



793 Actinomycin C.^ (VII) Cc4H9oOi6N,o 
235° (dec), [2W -321° ±lo"= 



red crystals, m.p. 232- 




383 Polypeptides and Related Compounds 

Below is shown one of the peptide side-chains of actino- 
mycin C3 to permit comparison with etamycin, etc. 



CH, 







CH 




Point of aftachn 


i 


\\ 




/ 
~^CH 


/ 


to actinocinii 


CH3-CH 

\ 


-0- 


NH 

/ 




CH 






i-Thre 


CH 




CH, / 


L-N-Me 
Vol 






\^^° 




/ 








\ 




o=c 

\ so 








NH 

/ 




re 








CH. 






D-0//0- 


P^ 




\ 






Isoleu 


/ ^ 


CH-CH,— CH 


N 




L-Pro 


C 


\ 


/ 


// 




\ 


\ 


CH3 


CH3 


-CH- 












CH, 


CH 


2 





" CH.. 

Streptomyces antibioticus, S. chrysomallus 
H. Brockmann, G. Bohnsack, B. Franck, H. Grone, H. Mux- 
feldt and C. Siiling, Angew. Chem. 68 70 (1956) and preced- 
ing papers. (Structure) 

H. Brockmann, W. Sunderkotter, K. W. Ohly and P. Boldt, 
Naturwissenschaften 47 230 (1960). 
' H. Brockmann and L. Lackner, ibid., 47 230 (1960). 

794 Actinomycin C^ (D,IV,Xi,Bi,Ii) C(.iHjj„Oip,Nio red prisms, m.p. 
241° (235.5-236.5) (dec.) [a],r" -349° ±10° (337°). 

\ 
^L-Thre — D-Val — L-Pro — Sar — L-Meval — O Indicates position 

of lactone not 

— L-Thre — D-Val — L-Pro — Sar — L-Meval — O , proved 



Where 

r \° NH, 

N 

CH3 ° 

Streptomyces chrysomallus, S. antibioticus; S. parvul- 
lus 

A. W. Johnson and A. B. Mauger, Biochem. J. 73 535 
(1959). 

Hans Brockmann and Hans-Sieghard Petras, Naturwis- 
senschaften 46 400 (1959). 



Pfizer Handbook of Microbial Metabolites 



384 



Hans Brockmann, P. Boldt and Hans-Sieghard Petras, 
ibid. 47 62 (1960). 

795 Actinomycin Co (VI) CeoHgoOieNjo red crystals, m.p. 237° (dec), 
[alD^' -325° ±10°. 



— L-Thre — D- Va I — L- Pro — Sa r — L-Meva I — O 



-L-Thre — D-0//0 — lieu — L-Pro — Sar — L-Meval — O 



\ 



Indicates it is not known in which chain the 
two acids ore. 



Streptomyces chrysomallus 

A. W. Johnson and A. B. Mauger, Biochem. J. 73 535 
(1959). 

Hans Brockmann and Hans-Sieghard Petras, Naturwis- 
senschaften 46 400 (1959). 

Hans Brockmann, P. Boldt and Hans-Sieghard Petras, ibid. 
47 62 (1960). 

C2a C62H90O26N12 an isomer of C2 found by paper 
chromatography. 

Streptomyces chrysomallus 

Hans Brockmann and B. Franck, ibid. 47 15 (1960). 

796 Actinomycin E^, C64H960i6Ni2- 



— Thre — a— lieu — Pro — Sar- 



Meval — O 



-Thre — a — lieu — Pro — Sar — Meileu — O 



Streptomyces sp. 

Giinther Schmidt-Kastner, Naturwissenschaften 43 131 
(1956). 

797 Actinomycin E2, C65H9„Oi6Ni2. 



-Thre — a — lieu — Pro — Sar — Meileu — O 
-Thre — a — lieu — Pro — Sar — Meileu — -O 



Streptomyces sp. 



385 



Polypeptides and Related Compounds 



Giinther Schmidt-Kastner, Naturwissenschaften 43 131 

(1956). 



798 Actinomycin Fp CngHssOieNjo. 



— Thre— 



Va I Sa r— Sa r Meva I — O 



— Thre — a — lieu — Sar — Sar — Meva! — O 



Streptomyces sp. 

Giinther Schmidt-Kastner, Naturwissenschaften 43 131 
(1956). 



799 Actinomycin Fo, CgoHg^^OifiNis. 



-Thre— Val- 



Pro Sar Meva I — O 



Thre — a — lieu Sar — Sar — Meval — O 



Streptomyces sp. 

Giinther Schmidt-Kastner, Naturwissenschaften 43 131 
(1956). 



800 Actinomycin F3, CsgHgoOjeNio- 

I 
-Thre — o — I leo — Sar — Sa r — Meva I — O 

-Thre — a — lieu — Sar— Sar — Meval — O 



Streptomyces sp. 

Giinther Schmidt-Kastner, Naturwissenschaften 43 131 
(1956). 

801 Actinomycin F4, CeiHgoOieNja. 

r— I 

— Thre — a — lieu — |Pro — Sar — Meval — O 

— ^Thre — a — lieu — Sar — Sar — Meval — O 



Streptomyces sp. 

Giinther Schmidt-Kastner, Naturwissenschaften 43 131 
(1956). 



Pfizer Handbook of Microbial Metabolites 



386 



802 Actinomycin Xi„ C59H87O17N] 



-Thre— Val- 



Oxopro — Sar — Meval — O 



Thre- Vol Sar — Sar — Meval — O 



Streptomyces chrysomallus, S. fradiae 

Hans Brockmann and H. Grone, Chem Ber. 87 1036 

(1954). 

803 Actinomycin X2 (V, Bo) CeiHggOiTNio red plates, m.p. 244-246°, 
[aW> -341° ±10°. 



-Thre — Val — Oxopro — Sar — Meval — O 



—Thre — Vol — Pro —Sar— Meval— O 



Streptomyces chrysomallus, S. fradiae 
Hans Brockmann and Hans Grone, Chem. Ber. 87 1036 
(1954). 

804 Actinomycin X3, needles. 

An actinomycin X3, containing threonine, sarcosine, 
proline, valine, isoleucine and N-methyl valine, also has 
been isolated. 

H. Brockmann and H. Grone, Chem. Ber. 87 1036 (1954). 

Werner Frommer, Arch. Mikrobiol. 34 1 (1959). 

805 Actinomycin -X^^ (I) C61H90O17N10 yellow needles, m.p. 245— 

247, [aW -260° ±10° (c 0.22 acetone). 



-Thre— Val- 



Hypro — Sar — Meval — O 



Thre Val Pro — Sar— Meval— O 



Streptomyces chrysomallus, S. fradiae 
Hans Brockmann, Gottfried Pampus and Jost H. Manegold, 
Chem. Ber. 92 1294 (1959). 

Hans Brockmann and H. Grone, Chem Ber. 87 1036 

(1954). 



387 Polypeptides and Related Compounds 

806 Actinomycin X,,,, C-.gHssOieNj.. 



— Thre— Val- 



Pro — Sar — Meval — O 



— Thre — Vol — Sar — Sar — Meval — O 



Streptomyces chrysomallus, S. fradiae 

Hans Brockmann and Gottfried Pampus, Angew. Chem. 67 

519 (1955). 

H. H. Martin and Gottfried Pampus, Arch. Mikrobiol. 25 

90 (1956). 

807 Actinomycin X„„ Cfi4H9oOi7Ni2. 

— Thre — Vol — p — Hypro — Sar — Meval — O 
Thre Vol Pro— —Sar— Meval — O 



Streptomyces chrysomallus, S. fradiae 
Same references as Actinomycin X„.^. 

Actinomycins Z. 

All contain the same five amino acids on hydrolysis: 
threonine, sarcosine, N-methylalanine, vaUne and N- 
methylvaline. 

808 Actinomycin Z,„ amorphous orange-red powder, m.p. 250° (dec. ). 

Streptomyces fradiae 

R. Bossi, R. Hiitter, W. Keller-Schierlein, L. Neipp and 
H. Zahner, Helv. Chim. Acta 41 1645 (1958). 

809 Actinomycin Z^, orange-red crystals, m.p. 256-260 (dec), [(x]v 

-362° (c 0.185 in CHCI3). 

Streptomyces fradiae 

R. Bossi, R. Hiitter, W. Keller-Schierlein, L. Neipp and 
H. Zahner, Helv. Chim.. Acta 41 1645 (1958). 

Actinomycins 'L<^,'L._<^,'L\, an inseparable mixture, m.p. 256-260° 
(dec), [a"]„ -246 (c 0.257 in CHCl,)- 

R. Bossi, R. Hiitter, W. Keller-Schierlein, L. Neipp and 
H. Zahner, Helv. Chim. Acta 41 1645 (1958). 



Pfizer Handbook of Microbial Metabolites 



388 



810 Actinomycin Z5, short red staffs, m.p. 261-267 (dec), [<x]v 

-284° (c 0.244 in CHCI3). 

Streptomyces fradiae 

R. Bossi, R. Hiitter, W. Keller-Schierlein, L. Neipp and 
H. Zahner, Helv. Chim. Acta 41 1645 (1958). 

811 Actinomycin II, Cr.^HseOigNio red plates, m.p. 215° [aln'^ -157° 

(c 0.24 in CHCI3). 



-L-Thre — D- Va I — Sar — Sa r — N — Meva I — O 
-L-Thre— D- Va I — Sa r — Sa r — N — Meva I — O 



Streptomyces chrysomallus 

A. W. Johnson and A. Mauger, Biochem. }. 73 535 (1959). 
WiUiam A. Goss and Edward Katz, Antibiotics and Chemo- 
therapy 10 221 (1960). 

812 Actinomycin III, CggHj^eOieNio red prisms, m.p. 237°, [air" 
-205° (c 0.22 in CHCI3)'. 



=— L-Thre — D-Vol- 



Sor— Sar— N— Me— Vol O 



L-Thre— D-Val—L-Pro —Sar— N — Me— Vol- O 



Streptomyces chrysomallus 

A. W. Johnson and A. Mauger, Biochem. J. 73 535 (1959). 
William A. Goss and Edward Katz, Antibiotics and Chemo- 
therapy 10 221 (1960). 



389 



Polypeptides and Related Compounds 



813 Mycobacillin, Co.-.HssOsoNi;^. colorless needles. 





O 




CH- 











\\ 




1 




^CH, 


CHj 


-C 




-CH— 


■NH / 


CHi 


\ 


r. \ ^ 


NH-^ 






C^ 


^ / 


CHj 


v.r 






Asp 




'V 


/ \ 


/C Ala 










Pro 1 


N 


HOOC\ / 












\^-/' .COOH 
\ /CH: 


CH Asp 












Asp CH'^ 


°-/ 












\ 


^C 












NH 


/ 
NH 

"°°'-"-CH,-c'h G,. 












\- 












01. 1-CH.-CH.-COOH 


0- 












T 


\ 

NH 

\ 

^\ ^CH Leu 
CH \ 












h 












Tyr CH^ 

/ CHj /f \ 
/ ^/ ^>— OH 


c/ oA 












NH \=/ 
/ 


NH 












cC 


\ Asp 












/\) 


CH 










Asp 


CH " 


/ \ 










X \ 


CH. X, 
/ q// ^NH 
HOOC ° 


Ser 

^'CH— . 

/ 


-c 

II 


NH 


Tyr 

^CH-^ 

\ 


NH 

c 


CH 

COOH 




CH2 




CHa 








/ 







\ 








OH 






OH 







Bacillus subtilis 

Hydrolysis yields five aspartic acids, two glutamic ac- 
ids, two tyrosines, one proline, one serine, one leucine and 
one alanine. (Unspecified configurations) 

S. K. Majumdar and S. K. Bose, Nature 181 134 (1958). 
(Isolation) 

Idem., Biochem. J. 74 596 (1960). (Structure) 



Pfizer Handbook of Microbial Metabolites 



390 



814 Bacitracin A, CeeHiogOigN^yS, white, hygroscopic, amorphous 
powder, [a]i,'' +5° (±2.5°). 











CH3CH: 


S— CHj 






\ 






\ 

CH— CH- 

/ 1 


/ 
-C 

\ 












CHa NH2 


N— CH 






CH2 COOH 
















\ / 








c=o 

1 






D-Asp CH 
















\ ^ 








NH 


CH3 




NH 













/ 




\/ 


II 






-Leu CH— CHr 


-CH 




CH 


,-— C- 


— -NH 


^CH. 


1 

c=o 

1 


\ 
CH, 




NH L-Asp 






l-Lys CH. 

CH, ^ 


1 

NH 

1 






0^/ 






-Glu CH— CH.- 


-CHi— COOH 


u^ r 


^C 
^CH.-^CH L-His 






\ 


1 

c=o 




HC 1 — 

1 1 






CH2 
CH— 

/ 








HN N 








NH 




V 

H 


NH 

ic 






NH-c_CH-CH- 


-CH,-CH3 








C^ 




CH3 






° \ D-Phe 
CH 

y^^"- NH L 






NH 
/ 




L-lleu 






-lleu 


D- 


Orn CH 










r^ ^c^ 






"K CH'^ 










// 


~CH — 

/ 


-NH^ 


V \„, 













CH 




\ 









CH3 



/ 



CH2 



CHo 



\ 



NH2 



CH3 



Bacillus subtilis, B. licheniforTnis 

The bacitracins are a difficultly separable polypeptide 
complex. Bacitracins A, B, C, D, E, F^, F^, F;^ and G have 
been differentiated. The F series may be artifacts. The 
structure of bacitracin A has received the most attention. 
In certain of the other bacitracins isoleucine is replaced 
by valine. The complex from B. liche7iiformis was origi- 
nally called ayfivin. 

I. M. Lockhart, E. P. Abraham and G. G. F. Newton, 
Biochem. J. 61 534 (1955). 

J. R. Weisiger, W. Hausmann and L. C. Craig, /. Am. Chem. 
Soc. 77 731, 3123 (1955). 

Dorothy Wrinch, Nature 179 536 (1957). 

E. P. Abraham, "CIBA Lectures in Microbial Biochemistry," 



39^ Polypeptides and Related Compounds 

The bacitracins, John Wiley and Sons, New York, 1957, pp. 
1-30. (A review which also covers the earlier work) 

815 Subtilin, amorphous white powder, [a],,-' —29° to —35° 

Subtilin is a basic polypeptide, molecular weight 3188, 
which yields 11 common amino acids, lanthionine: 

HOOC— CH— CH2— S— CH.— CH— COOH 
NH.> NH^ 

and a new S-amino acid, probably yS-methyllanthionine : 
(0 CH3(d) 

HOOC— CH— CH.— S— CH— CH— COOH 
NH. NH., 

The common amino acids identified are: glycine, ala- 
nine, valine, leucine, isoleucine, proline, phenylalanine, 
tryptophan, lysine, asparagine and glutamic acid. 
Bacillus subtilis 

Eugene F. Jansen and Doris J. Hirschmann, Arch. Biochem 
4 297 (1944). 

A. J. Salle and Gregory J. Jann, Proc. Soc. Exp. Biol. 60 60 
(1945). 

W. Steenken, Jr. and E. Wolinsky, /. Bad. 57 453 (1949). 

J. C. Lewis and N. S. Snell, /. Am. Chem. Soc. 73 4812 
(1951). 

Gordon Alderton, ibid. 75 2391 (1953). 

Nisins, nearly white needles. 

Consist of four active cyclic polypeptides. All contain 
lanthionine and yS-methyllanthionine. These amino acids 
also occur in the antibiotics, subtilin, cinnamycin and 
duramycin. 

816, 817, 818 Nisins A, B and C contain leucine and/or isoleucine, val- 
ine, alanine, glycine, proline, aspartic acid, histidine, 
lysine and methionine. 

819 Nisin D contains glutamic acid, but no valine or methionine. 

Nisin A has a molecular weight of --7000 and also 
I contains serine. 

I Streptococcus lactis, S. cremoris 

N. J. Berridge, G. G. F. Newton and E. P. Abraham, 
Biochem. J. 52 529 (1952). 
I G. G. F. Newton and E. P. Abraham, Nature 171 606 (1953). 



Pfizer Handbook of Microbial Metabolites 392 

G. Cheeseman and N. Berridge, Biochem. J. 71 185 (1959). 

820 Duramycin, colorless amorphous soUd, no definite m.p., Hydro- 

chloride: [a]rr' -6.4° (c 3.9 in water). 

Duramycin is a polypeptide, containing at least one 
free amino group and several free carboxyl groups. Acid 
hydrolysis yielded: lanthionine, ^-methyllanthionine, as- 
partic acid, glutamic acid, glycine, valine, proline, phenyl- 
alanine and possibly ornithine and hydroxyproUne. Du- 
ramycin is related to, but distinct from, cinnamycin. 
Streptomyces cinnamoneus f . azacoluta 
Odette L. Shotwell, Frank H. Stodola, William R. Michael, 
Lloyd A. Lindenfelser, Robert G. Dworschack and Thomas G. 
Pridham, J. Am. Chem. Soc. 80 3912 (1958). 

821 Cinnamycin. 

A polypeptide containing (probably): glutamic acid, 
aspartic acid, proline, phenylalanine, valine, arginine, 
lanthionine and /3-methyllanthionine. 

Streptomyces cinnamoneus 

Robert G. Benedict, William Dvonch, Odette L. Shotwell, 
Thomas G. Pridham and Lloyd A. Lindenfelser, Antibiotics 
and Chemotherapy 2 591 (1952). 

Robert G. Benedict, Bot. Rev. 19 229 (1953). 

822 Matamycin, colorless crystals, m.p. 173° (dec), [ajn^" +36.6° 

(c 0.11 in methanol). 

An essentially neutral antibiotic of low solubility. 
Analysis: C 43.95, H 4.06, N 14.45, S 13.57. Halogen- 
free. Positive Fehlings, ToUens, permanganate, DNPH, 
and (after hydrolysis) ninhydrin tests. Negative ferric 
chloride and Sakaguchi tests. A hydrolysate contained: 
cysteic acid, glycine, serine, alanine, arginine and two 
other amino acids. 

Streptomyces matensis n. sp. 

P. Sensi, R. Ballotta and G. G. Gallo, Antibiotics and Chem- 
otherapy 9 76 (1959). 

An inactive compound, "Compound I," evidently of 
analogous structure was isolated from the same culture: 

823 Compound I, colorless crystals, m.p. 189° (dec), [aln'" +151.6° 

(c 0.1 in dioxane). 

Analysis: C 45.84, H 3.90, N 14.99, S 14.64. It may 
be a dehydration product of matamycin. 



393 Polypeptides and Related Compounds 

824 Comirin, nearly colorless powder, m.p. 230-235° (dec.)- 

A polypeptide containing the following amino acids: 
serine, aspartic acid, glycine, a,y-diaminobutyric acid, 
lysine, leucine, isoleucine, tyrosine and arginine. An 
ether-soluble moiety also was present. Negative ninhy- 
drin, positive biuret. No free amino acid groups. 
Pseudomonas antimycetica 
W. G. C. Forsyth, Biochem. J. 59 500 (1955). 

825 Colimycin. 

A crystalline polypeptide, containing mainly D-leucine 
and L-threonine. 
Bacillus colistinus 
P. V. Forni and E. Guidetti, Minerva med. II 823 (1956). 

826 Brevin. 
Brevin is a polypeptide containing: aspartic acid, gly- 
cine, tyrosine, serine, an unidentified basic substance 
(and also a fatty acid component?). 

Bacillus brevis 

I Ella M. Barnes and G. G. F. Newton, Antibiotics and Chem- 

otherapy 3 866 (1953). 

827 Brevolin, Hydrochloride yellowish white amorphous, [ajn^*' 

-18.9°. 

Brevolin is a polypeptide, probably related to brevin. 

Bacillus brevis 

Junichi Kawamata and Yutaka Motomura, J. Antibiotics 
(Japan) 7A 25 (1954). 

Antibiotics from Yeast. 

Two amorphous compounds have been isolated from 
bakers' yeast. They have antibacterial and antifungal 
effects, and seem to be cyclic polypeptides. Acid hydroly- 

828 sis of one of these (Y^) gave leucine, valine, alanine, 

829 glutamic acid and glycine. Acid hydrolysis of Y2 gave the 
same amino acids plus y-aminobutyric acid. 

■ Werner Motzel and Elton S. Cook, Nature 182 455 (1958). 

F ,, . 

830 Alvem. 

A basic polypeptide containing arginine. 
Bacillus alvei 

K. Gilliver, A. M. Holmes and E. P. Abraham, Brit. J. 
Exptl. Path. 30 209 (1949). 



Pfizer Handbook of Microbial Metabolites 394 

831 Thiostrepton, colorless crystals, m.p. 246-256° (dec), [ajo"^ 

—98.5° (c 1 in glacial acetic acid). 

A weakly basic polypetide. Probable amino acid con- 
tent: leucine (or isoleucine), valine, alanine, threonine, 
proline, lysine, glycine, aspartic acid, glutamic acid, 
cystine and tryptophan. 

Streptomyces sp. 

John Vandeputte and James D. Dutcher, "Antibiotics An- 
nual 1955-1956," Medical Encyclopedia, Inc., New York, p. 
560. 

832 Antibiotic 899, reddish yellow amorphous powder, m.p. 115- 

120°. 

A neutral compound with spectra similar to those of 
streptogramin. 

Streptomyces sp. resembling S. virginiae 
P. De Somer and P. Van Dijck, Antibiotics and Chemother- 
apy 5 632 (1955). 

833 Amphomycin, colorless crystals, [ajn"' +7.5° ±5 (c 1 in water 

at pH 6). 

An acidic (amphoteric) polypeptide, minimal molecu- 
lar weight about 1500. 

Streptomyces canus 

Bernard Helnemann, Murray A. Kaplan, Robert D. Muir 
and Irving R. Hooper, Antibiotics and Chemotherapy 3 1239 
(1953). 

834 Aspartocin. 

An acidic polypeptide similar to amphomycin. C 53.2, 
H 7.6, N 13.2, S 0.42, no halogen. Hydrolyzes to 4 
moles of L-aspartic acid, 2 moles of glycine, 1 mole of 
L-proline, 1 mole of L-valine, a,y8-diaminobutyric acid, 
a-lL],/8-methylaspartic acid, D-a-pipecoHc acid and an un- 
saturated fatty acid. 

Streptomyces griseus var. spiralis, S. violaceus 

Yields of 1 to 10 g. per liter were obtained. 

A. J. Shay, J. Adam, J. H. Martin, W. K. Hausmann, P. Shu 
and N. Bohonos, 7th Annual Symposium on Antibiotics, Wash- 
ington, D. C, 1959. 

J. H. Martin and W. K. Hausmann. J. Am. Chem. Soc. 82 
2079 (1960). 

835 Zaomycin, m.p. 242-246° (dec). 

A polypeptide resembling amphomycin. 
Streptomyces zaomyceticus n. sp. 



395 Polypeptides and Related Compounds 

Yorio Hinuma, /. Antibiotics (Japan) 7A 134 (1954>. 

836 Bacillomycin (Fungocin, Bacillomycin R, Bacillomycin A), color- 

less microcrystals. 

An acidic polypeptide, molecular weight ~1000. Anal- 
ysis: C 52.69, H 7.20, N 12.29. Contains glutamic acid, 
aspartic acid, serine, threonine and tyrosine. Similar to 
or identical with eumycin. 

Bacillus siibtilis 

Maurice Landy, Sanford B. Rosenman and George H. War- 
ren, ;. Bad. 54 24 (1947). 

Howard Tint and Wilhelm Relss, 7- Biol. Chem. 190 133 
(1951). 

Robert A. Turner, Arch. Biochem. 60 364 (1956). 

837 Bacillomycin B, amorphous yellow material. 

A polypeptide containing glutamic acid, aspartic acid, 
proline, tyrosine and leucine. 
Bacillus subtilis 

Isao Shibasaki and Gyozo Terui, /. Fermentation Technol. 
(Japan) 31 339 (1953). 

838 Bacillomycin C. 

A polypeptide containing glutamic acid, aspartic acid, 
tyrosine, leucine and valine. 

Bacillus subtilis 

Isao Shibasaki and Gyozo Terui, /. Fermentation Technol. 
(Japan) 32 115 (1954). 

839 Fungistatin. 

An amphoteric polypeptide, containing aspartic acid, 
lysine, serine, threonine, proline, alanine, isoleucine, 
valine, tryptophan, tyrosine, other unidentified amino ac- 
ids. Molecular weight about 2400. 

Bacillus subtilis 

Gladys L. Hobby, Peter P. Regna, Nancy Dougherty and 
WiUiam E. Steig, J. Clin. Invest. 28 927 (1949). 

P. P. Regna, R. A. Carboni and W. E. Steig, Am. Chem. Soc. 
Meeting-in-Miniature, Brooklyn (1950). 

Robert L. Peck and John E. Lyons, Ann. Rev. Biochem. 20 
367 (1951). 

840 Bryamycin, m.p. 223-235° (dec), [«]«'' -68.5° (c 1 in chloro- 

form ) . 

A polypeptide containing alanine, glycine, isoleucine, 
threonine, cystine and unidentified compounds. 



Pfizer Handbook of Microbial Metabolites 396 

Streptomyces hawaiiensis n. sp. 

M. J. Cron, D. F. Whitehead, I. R. Harper, B. Heinemann 
and J. Lain, Antibiotics and Chemotherapy 6 63 (1956). 

841 Coliformin. 

A polypetide, molecular weight 4000 ± 400, containing 
glutamic acid, aspartic acid, lysine, valine, leucine, serine, 
alanine and glycine. Positive Molisch. Contains traces 
of phosphorus and sulfur. 

An E. coli-Aerobacter aerogenes type of bacterium 
Stig K. L. Freyschuss, Stig O. Pehrson and Borje Steinberg, 
Antibiotics and Chemotherapy 5 218 (1955). 

842 Mycosubtilin, white crystals, m.p. 256°. 

A polypeptide, C 55.31, H 7.61, N 15.15. 
Bacillus subtilis 

Robert P. Walton and H. Boyd Woodruff, /. Clin. Invest. 28 
924 (1949). 

843 Grizein ( Helianthate ) homogeneous brown powder, m.p. 194- 

196° (dec.) (hydrochloride) white, hygroscopic powder. 

A basic polypeptide complex. Positive biuret, ninhy- 
drin, glucosamine reactions. Negative maltol, histidine, 
Sakaguchi tests. 

Streptomyces griseus-like strains 

N. A. Krasilnikov, A. N. Belozerskii, Ya. I. Rautenshtein, 
A. I. Korenyako, N. I. Nikitina, A. I. Sokolova and S. O. Ury- 
son, Mikrobiologiya 26 418 (1957). 

Licheniformins, amorphous white powders, no m.p. 

844 Licheniformin A, hydrochloride: [aln^^ —37.4° (c 1 in chloro- 

form). 

845 Licheniformin B, hydrochloride: [a]D^° —37.7° (c 1 in chloro- 

form). 

846 Licheniformin C, hydrochloride: [ajn^" —36.8° (c 1 in chloro- 

form ) . 

A rather high molecular weight polypeptide complex. 
Negative glucosamine and Molisch. Positive Sakaguchi, 
biuret. 

Licheniformins A and B contain: aspartic acid, glycine, 
serine, lysine, arginine, valine, proline and phenylalanine. 

Bacillus licheniformis 

R. K. Callow, R. E. Glover, P. D'Arcy Hart and G. M. Hills, 
Brit. J. Exptl. Path. 28 418 (1947). 



397 Polypeptides and Related Compounds 

R. K. Callow and T. S. Work, Biochem. J. 51 558 (1952). 

847 Carcinomycin, dark green, amorphous. 

A polypeptide antibiotic. Sulfur-free. 
Streptornyces carcinomyciciis 

Shogo Hosotani and Momoe Soeda, Japanese Patent 6893 
(1959). (Chem. Abstr. 54 831g) 

848 Carcinocidin, [oc]u'^ —20° (c 1 in water). 

A polypeptide antibiotic, containing cystine, lysine, 
glycine and glutamic acid. Molecular weight >6000. 

Streptornyces kitazazvaensis 

This organism also produces antimycin A. 

F. Okamoto, Shigeo Kubo, Takahashi Nara and Shiro Ta- 
naka, Jap. Patent Appl. 6894 (1959). (Chem. Abstr. 54 832c) 

849 Melanomycin (Sodium Salt), brown, amorphous powder. 

A polypeptide antibiotic yielding on hydrolysis : phenyl- 
alanine, leucine, valine, proline, alanine, glutamic acid 
and histidine. 

Streptornyces melanogenes 

Fujiki Hata, Ryozo Sugawara, Akihiro Matsumae and Taka- 
moto Sano, Japanese Patent 5899 (1959). {Chem. Abstr. 54 
833b) 

850 Notatin (Penicillin B, Penatin), buff colored powder, water sol- 

uble, [ajn'" -4.8° (c 0.012 in water). 

A fiavoprotein enzyme (glucose-oxidase), molecular 
weight about 152,000. 

PenicilliuTn notatum, other Penicillium spp. 

R. Cecil and A. G. Ogston, Biochem. J. 42 229 (1948). 




18. 



Heterocycles 



a. FURANS AND RELATED SUBSTANCES 

Apparently there has been no investigation of the biosynthetic 
origin of the furans listed here, but it is known that furans can 
be formed in several different ways. 

The relationship of furans to sugars is recognized in the desig- 
nation of the five-membered ring hemi-acetal form of sugars as 
the furanose form. Dilute acid converts glucose to 5-hydroxy- 
methylfurfural. The latter compound may be a precursor of 
Sumiki's acid, although the transformations are probably enzy- 
matic. The four carbon atom sugar erythrose also is a likely 
furan precursor as pointed out by Wenkert.^ 

The furans with carbon chains at the 2-position are obviously 
terpenoid. Since they were isolated from a sweet potato me- 
dium, their direct derivation from glucose cannot be assumed. 
The simpler substances may arise from oxidation of the more 
complex. 

It is interesting to note that the lactone side-chain of digitoxi- 
genin is derived from acetate rather than from mevalonic acid.^ 
Such lactones as well as the related tetronic acids, would seem 




Digitoxigenin 



HO 

to be potential furan precursors. 

1 Ernest Wenkert, Experientia 15 165 (1959). 

^ E. Leete, Seventh Medicinal Chemistry Symposium of the Amer- 
ican Chemical Society, Kingston, Rhode Island, 1960. 



399 Furans and Related Substances 

851 Furan-3-carboxylic Acid, C.-,H40;,, colorless crystals, m.p. 121°. 

COOH 

/ 



u 



Ceratostomella fimbriata (sweet potato substrate) 
Takashi Kubota and Keizo Naya, Chem. and Ind., 1427 
(1954). 

852 5-Hydroxymethylfuran-2-carboxylic Acid (Sumiki's Acid), CeHr,04, 
colorless crystals, m.p. 164° (dec). 



I J 
HOCH, COOH 

Aspergillus glaucus, A. clavatus, A. niger, A. oryzae, A. 
wentii, Gibberella fujikuroi 

Yusuke Sumiki, J. Agr. Chem. Soc. Japan 7 819 (1931). 

Akira Kawarada, Nobutaka Takahashi, Hiroshi Kitamura, 
Yasuo Seta, Makoto Takai and Saburo Tamura, Bull. Agr. 
Chem. Soc. (Japan) 19 84 (1955). 

853 Ipomeanine, CoHi.Ai, oil, b.o.ooi 74-79°, Hd'-^ 1.4975, [a]o +3.9°. 

O O 

II II 

C— CH2— CH2— C— CHs 

Ceratostomella fimbriata (sweet potato substrate) 
Takashi Kubota and Nobutaka Ichikawa, Chem. and Ind., 
902 (1954). 

854 Batatic Acid, C10H10O4, colorless crystals, m.p. 88.5°, [ajc^" 

+ 17.5" (in ethanol). 




Ceratostomella fimbriata (sweet potato substrate) 
Takashi Kubota and Keizo Naya, Chem. and Ind., 1427 
(1954). 



Pfizer Handbook of Microbial Metabolites 400 

855 Ipomeamarone, C15H20O3, colorless oil, b. 0.001 103°, ric^^ 1.4827, 
[<x]u-' +28°. 

-CH CH2 

1 I 

O CH2 



U 



CH3 C— CHo— CH2— CH 



CH3 



CH3 



Ceratostomella fimbriata (sweet potato substrate) 

T. Kubota and T. Matsuura, Chem. and Ind., 521 (1956). 
(Synthesis) 

There is a marked resemblance between ipomeamarone 
and dendrolasin, an oil Ci.r.HoL.O, isolated from ants. It is 
an enantiomer of ngaione, isolated from Myoporum spp. 
(higher plant). 

A. Quilico, F. Piozzi and M. Pavan, Tetrahedron 1 177 
(1957). (Structure) 

A. J. Birch, R. A. Massy-Westropp and S. E. Wright, Chem. 
and Ind., 902 (1954). 

Ipomeamarone is thought to be formed by the host 
(sweet potato) tissue to resist invasion by Ceratostomella 
fimbriata.* 

b. DIBENZOFURANS AND RELATED SUBSTANCES 

Dibenzofurans constitute a class of natural products 
found only in lichens. Usnic acid is the most widely dis- 
tributed dibenzofuran. Its structure, which was contro- 
versial for some time, now has been proved by synthesis.^ 
The dibenzofurans are formed from 2 moles of the ace- 
tate-derived resorcinolic substances typical of lichens. 
Results of chemical experiments, including the method of 
synthesis of usnic acid, make it quite probable that phenol 
coupling of the sort mentioned in connection with dep- 
sides and depsidones also is involved here.-' ^ Thus, 

* T. Akazawa, Arch. Biochem. and Biophys. 90 82 (1960). 

1 D. H. R. Barton, A. M. Deflorin, O. E. Edwards and J. B. Hen- 
drickson, Chem. and Ind., 1670 (1955). 

2 D. H. R. Barton and T. Cohen, Festschr. Arthur Stoll, 117 (1957). 
^ Holger Erdtman and Carl Axel Wachtmeister, ibid., 144 (1957). 



401 



Dibenzofurans and Related Substances 



dydymic acid would be formed by coupling of two similar 
orsellinic acids: 



CH3CH,CH2 ^^^^ CHaCH,CH,CH,CH, ^^^^ 
I COOH I COOH 



+ 



HO 



OH 



HO 




OH 



CH3CH2CH2 



P^ 




OH 

I 
CO 



CH2CH2CH2CH2CH 



0|0 
H 




COOH 



OH 



HvCsl^^ CaHu^QQ^ 



HO 




H7C3 



C5H11 



H7C3 



CsHii 



COOH 







HO 



And in the case of usnic acid: 



COOH 



OH CH3O O OH 

Didymic Acid 





H H 

O O 

CH3I COCH3 CH3 



+ 



HO I '-^ O 

COCH3 H 




%. 




HO I OH O 

COCH3 H 

i 
H H H H 

00 00 

CH3 I CH,! COCH3 CH3 I r»J COCH3 

" ' — H2O ^ ' " ' 





COCH3 

Usnic Acid 



HO I O 

COCH3 H 



Pfizer Handbook of Microbial Metabolites 



402 



Formation of the monobenzofuran shown also may in- 
volve phenol coupling, if not precisely as indicated at least 
in the same general fashion: 




Apparently, many lichens contain an enzyme system 
which can promote phenolic coupling of this type. Nei- 
ther the dibenzofurans nor the depsides and depsidones 
are produced by molds alone (although some of their 
resorcinolic precursors are), and the algal partners must 
be required in the coupling process. 

856 Strepsilin, C15H10O5, colorless crystals, m.p. 324°. 




Cladonia strepsilis Wain. 

Shoji Shibata, 7- Pharm. Soc. Japan 64 20 (1944). 
ture) 



( Struc- 



403 Dibenzofurans and Related Substances 

857 Porphyrilic Acid, C,,;H,o07, colorless needles, m.p. 280-283° 

(dec). 

O—CH... OH 

CH.> X I 

HO ' ^"3 

COOH 

Haematomma coccineum (Dicks.), H. porphyrium 
(Pers.) 

Porphyrilic acid occurs together with Z-usnic acid and 
atranorin. 

Carl Axel Wachtmeister, Acta Chem. Scand. 10 1404 
(1956). (Structure) 

858 2-(6-Hydroxy-2-inethoxy-3,4-methylenedioxyphenyl)-benzofuran, 

CieHioOg, colorless crystals, m.p. 118°. 




Yeast 

A yield of 0.5-2.0 mg. per pound of bakers' yeast was 
reported. 

M. A. P. Meisinger, Frederick A. Kuehl, Jr., E. L. Rickes, 
Norman G. Brink, Karl Folkers, Martin Forbes, Friederich 
Zilliken and Paul Gyorgy, J. Am. Chem. Soc. 81 4979 (1959). 
(Structure) 

859 Pannaric Acid, CigHioO;, colorless needles, m.p. 243-245°. 

OH CH3 ^^^^ 

COOH 




COOH 



Crocynea membranacea (Dicks.) Zahlbr. = Pannaria 
lanuginosa Ach. 

O. Hesse, /. prakt. Chem. 70 1 (1904). (Isolation) 



Pfizer Handbook of Microbial Metabolites 404 

Akermark H. Erdtman and C. A. Wachtmeister, Acta Chem. 
Scand. 13 1855 (1959). (Structure) 

860 d- and MJsnic Acid, CjsHieOj, yellow crystals, m.p. 203°, [ixW 
(d-form) +492°, (Z-form) -495°. M.p. d,Z-form 195°. 




CH3 

Usnea, Alectoria, Ramalina, Evernia, Cetraria, Parmelia, 
Cladonia, Lecanora and Haematormna species (most yel- 
low lichens). Long known. 

Both d- and Z-forms occur in lichens. Relatively high 
yields are available from some species. 

Clemens Schdpf and Friedrich Ross, Naturivissenschaften 
26 772 (1938). 

Idem., Ann. 546 1 (1941). (Structure) 

D. H. R. Barton, A. M. Deflorin, O. E. Edwards and J. B. 
Hendrickson, Chem. and Ind., 1670 (1955). (Synthesis) 

861 Didymic Acid (Incrassatic Acid), C22H26O5, colorless crystals, 
m.p. 172°. 

C3H7 C5H11 _^_,, 

I I COOH 




CH3O OH 

Cladonia species (occurs together with squamatic and 
barbatic acids) 

Yasuhiko Asahina and Masaru Aoki, /. Pharm. Soc. Japan 
64 41 (1944). 

C. PYRANS AND RELATED SUBSTANCES 
The y-Pyrones and Patulin 

The biosynthesis of patulin was discussed in the intro- 
duction to the chapter on phenolic substances. 

Kojic acid has long attracted interest because it is pro- 
duced in such high yields by certain Aspergillus species. 
Within the past few years isokojic acid and several other 
related y-pyrones have been isolated from Gluconoaceto- 
bacter cultures. 



405 Ppans and Related Substances 

The fungi are able to use pentose and triose substfates 
as well as glucose, although labeling studies have shown 
conversion of glucose to kojic acid without cleavage of 
the 6-carbon chain. ^ 

Gluconoacetobacter liquefaciens seems to be more se- 
lective in its substrate and uses only glucose, gluconate 
and 2-ketogluconate. The variety of y-pyrones produced 
is useful in deducing the kind of intermediate involved. 
The foregoing considerations plus the isolation of 2,5- 
diketogluconic acid from cultures of this bacterium have 
led to formulation of the following biosynthetic route to 
the pyrones produced by Gluconoacetobacter liquefaciens:'^ 

CH2OH CH2OH 

I I 

c — o c — o 

H /I \ H H /I \ 

1/ H \i -2H IX H \ +H2O 

c c > c c=o > 

|\ OH H /| |\ OH H 

HO \| 1/ OH HO \| 

c — c c — c 

II II 

H OH H OH 

Glucopyranose Gluconolactone 

CH2OH 

I 

C— OH 
H yi OH 

IX H I -2H 

,C c=o 

^\\ OH 
-2H HO \| 

C— 

I 

H O 
2-Ketogluconic Acid 




H OH 
Gluconic Acid 



H OH 
5-Ketogluconic Acid 

1 H. R. V. Arnstein and R. Bentley, Biochem. J. 62 403 (1956). 
^ Ko Aida, Mitsuko Fujii and Toshinobu Asai, Bull. Agr. Chem. 
Soc. (Japan) 21 30 (1957). 



Pfizer Handbook of Microbial Metabolites 



406 



HO 



HO 



CH2OH 

I 

c=o 

OH 

J 

c — c 



OH 



c=o 



H O 

2, 5-Diketogluconic Add 



o=c 



CHOH 



C OH 

H / OH 



— 2H^ 
— H2O 



c=o 



OH 

C— C 

OH 
Enol Form 



-2H,0 



o=c 



OH H 

|- I 



OH H 

1 I 

c=c 



OH COOH 

Rubiginic Acid 

— CO2 

OH H 

I I 



c=c 

1 I 

OH H 

Rubiginol 



o=c 



c=c 

1 1 

H COOH 
Comenic Acid 



407 



Pyrans and Related Substances 



Another bacterial species, Gluconoacetobacter roseum, 
studied by the Japanese, produces kojic and isokojic acids 
and only from a fructose, sucrose or mannitol substrate. 
The two products are always found together. The pro- 
posed route by which these two pyrones are formed from 
fructose by Gluconoacetobacter roseum is shown below :^ 

CH:OH CH2OH CHOH CHO 



HOCH 



HOCH 

j 

HCOH 

HCOH 

CH.OH 
Mannitol 



CO 

2H HOCH 

HCOH 

HCOH 

CH2OH 
Fructose 



COH CO 

I I 

HOCH — 2H HOCH 

HCOH HCOH 



HCOH 

CH2OH 
Enol Form 



HCOH 

CHjOH 
Glucosone 



CHOH 

I 
COH 

II 
COH 

I 
HCOH 

HC 



CH2OH 




CH2OH 



CH2OH CH2OH 

Kojic Acid Isokojic Acid 

Kojic acid is potentially an inexpensive chemical be- 
cause of high yields from aspergilH. 

862 Rubiginol, C5H4O4, colorless plates, m.p. 203.5°. 

O 



HO 



OH 



^O 

Gluconoacetobacter liquefaciens 

A yield of 1.2 g. of rubiginol from 140 g. of glucose sub- 
strate was reported. 

Ko Aida, /. Gen. and Appl. Microbiol. (Japan) 1 30 (1955). 

^ Ko Aida, Mitsuko Fujii and Toshinobu Asai, Proc. Japan Acad. 
32 595 (1956). 



Pfizer Handbook of Microbial Metabolites 408 

863 Comenic Acid, CgH^Oj, colorless plates, m.p. 276° (dec.)- 

HO ? 



COOH 

Gluconoacetobacter liquefaciens 

A yield of 1.1 g. from 140 g. of glucose has been re- 
ported. 

Ko Aida, Bull. Agr. Chem. Soc. (Japan) 19 97 (1955). 



864 Rubiginic Acid, C6H4O6, colorless needles, m.p. 230° (dec). 

) 

V 



HO II OH 





COOH 



Gluconoacetobacter liquefaciens 

Ko Alda, Bull. Agr. Chem. Soc. (Japan) 19 97 (1955). 

865 Kojic Acid, C6H6O4, colorless prisms, m.p. 152°. 

HO ? 



CH2OH 



Aspergillus fiavus, A. oryzae, A. tamarii, A. glaucus, 
Gluconoacetobacter roseum (fructose substrate) 

High yields (45 g. or more per 100 g. of glucose sub- 
strate) are produced by some aspergillus strains. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
1954 Vol. "II, Lewis B. Lockwood, Ketogenic fermentation proc- 
esses, chap. 1, pp. 19-20. (A review) 

Andrew Bielik, Advances in Carbohydrate Chem. 11 145 
(1956). (A review) 

866 Isokojic Acid, C6H6O4, colorless plates, m.p. 183°. 

O 




HO CH2OH 



409 Pyrans and Related Substances 

Gliiconoacetobacter roseum (fructose substrate) - 
Isokojic acid was produced together with kojic acid and 

an unidentified substance. 

Ko Aida, Mitsuko Fujii and Toshinobu Asai, Proc. Japan. 

Acad. 32 600 (1956). 

867 Patulin (Clavacin, Clavatin, Claviformin, Penicidin, Expansine, 
Mycoin), C7H6O4, colorless crystals, m.p. 111°. 

O 



o 

I 




OH 

Penicillium patuhnn Bainier (P. urticae), P. griseo- 
fulvum, P. claviforme, P. expansum, P. melinii, P. equi- 
num, P. novae-zeelandiae, P. leucopus, Aspergillus clava- 
tus, A. terreus, A. giganteus, Gymrioascus spp. 

H. W. Florey, E. Chain, N. G. Heatley, M. A. Jennings, 
A. G. Sanders, E. P. Abraham and M. E. Florey, "Antibiotics," 
Oxford University Press, London, 1949, pp. 223-272. (Re- 
views earher work) 

R. B. Woodward and Gurbakhsh Singh, J. Am. Chem. Soc. 
72 1428 (1950). (Synthesis) 

868 5-Hydroxy-2-methylchromone, CioHgOa, yellow needles, m.p. 72- 
76°. 
Proposed structure : 

O 




CH3 
Daldinia concentrica 
D. C. Allport and J. D. Bu'Lock, J. Chem. Soc, 654 (1960). 

869 5-Hydroxy-2-niethylchromanone, C10H10O3, pale yellow needles, 
m.p. 30-33°. 

OH O 




CH3 
Daldinia concentrica 
D. C. AUport and J. D. Bu'Lock, /. Chem. Soc, 654 (1960). 



Pfizer Handbook of Microbial Metabolites 410 

870 Aureothin, C20H23O6N, yellow crystals, m.p. 158°. 

CH3 O 

OzH—^y-QH^C—CH^C CH2' ' 




OCH3 



Streptomyces thioluteus 

Aureothin occurs as a by-product in the aureothricin 
fermentation. 

Kenji Maeda, /. Antibiotics (Japan) 6A 137 (1953). (Iso- 
lation) 

Y. Hirata, H. Nakata and K. Yamada, /. Chem. Soc. Japan 
79 1390 (1958) and preceding papers. (Structure) 

QuiNONOiD Compounds. 

This section includes a group of colored compounds, 
many of which have chromophores resembling those of 
quinones. These unusual substances presented some 
interesting structural problems. In many cases there 
was a long time interval between isolation and complete 
structure determination. 

The relationship between fulvic acid and citromycetin 
is obvious. The relationship of both of these compounds 
to fusarubin has been pointed out recently.^ This is less 
obvious, but a precursor such as (I) was envisaged for all 
three compounds, the formation of fusarubin involving 
ring closure at the dotted line. 




Penicillium griseofulvum, which is one of the producers 

^ F. M. Dean, R. A. Eade, R. A. Moubasher and A. Robertson, Na- 
ture 179 366 (1957). 



411 Pyrans and Related Substances 

of fulvic acid, also produces a variety of other metabolites, 
including griseofulvin and mycelianamide. There seems 
to be no close relationship between these compounds and 
the three mentioned above, however. 

The biosyntheses of sclerotiorin,-' citromycetin- and cit- 
rinin-' -^ have been investigated by using C'^-labeled ace- 
tate, formate and methionine. 

The two studies of citrinin (III) were in agreement, the 
results of both indicating origin from a 10-carbon atom 
polyketomethylene chain in the sense of (II). 



C COOH 

/ \ / 11 OH 
CH2 CH.2 HOOC 1 , 

+3 C-atoms \i^;^'^'^3-^^^r\ 
> I \ ^ 


ti II ^ Q 

c I I o^VSAch 

/ \ / \ / \ CH3 CHs ,0" 
CH2 CH- CH3 12 13 



The carbon atoms 11, 12 and 13 in (III) were contrib- 
uted by methionine or formate. 

Sclerotiorin also is acetate derived with contribution of 
three carbon atoms by formate. 

Citromycetin (V) is derived entirely from seven acetic 
acid units, CH3— COOH (CH3— COOH) and yields the 
labeUng pattern shown below. 




It would appear that purpurogenone should also be de- 
rived from seven acetate units. 

- A. J. Birch, P. Fitton, E. Pride, A. J. Ryan, Herchel Smith and 
W. B. Whalley, /. Chem. Soc, 4576 (1958). 

^ Erwin Schwenk, George J. Alexander, Allen M. Gold and Dean F. 
Stevens, ;. Biol. Chem. 233 1211 (1958). 



Pfizer Handbook of Microbial Metabolites 



412 



871 Radicinin,* CjoHioOa, yellow crystals, m.p. 220° (dec), [aW 
-217.4° (c "2.37 in pyridine). 
Proposed partial structure: 



— -CH3 

=0 -'" 



Stemphylium radicinum Sterad (formerly Alternaria 
radicina) 

D. D. Clarke and F. F. Nord, Arch. Biochem. and Biophys. 
59 269, 285 (1955). 

872 Citrinin, C13H14O5, long yellow prisms, m.p. 179° (dec), [aW 
-34.5° (c 0.60 in alcohol). 



HOOC 





Penicillium citrinum, P. expansum, P. implicatum, 
P. chrzaszszi, P. citreo-sulfuratum, P. lividum, P. phaeo- 
janthinellum, Aspergillus terreus, A. candidus, A. niveus 

A. C. Hetherington and H. Raistrick, Trans. Roy. Soc. (Lon- 
don) B220 269 (1931). (Isolation) 

D. H. Johnson, Alexander Robertson and W. B. Whalley, 
J. Chem. Soc, 2971 (1950). 

H. H. Warren, Gregg Dougherty and Everett S. Wallis, 
J. Am. Chem. Soc. 71 3422 (1949). (Synthesis) 

873 Citromycetin (Frequentic Acid), C14H10O7, lemon yeUow hy- 
drated needles, m.p. 283-285° (dec). 




See entry 413. 



413 Pyrans and Related Substances 

Peuicillium frequentans, P. roseo-purpurogenum, P. gla- 
brum, P. pfefferianuvi, Citromyces strains, Corynebacte- 
rium diphtheriae 

A. C. Hetherington and H. Raistrick, Trans. Roy. Soc. (Lon- 
don) B220 209 (1931). (Isolation) 

Alexander Robertson, W. B. Whalley and J. Yates, /. Chem. 
Soc, 2013 (1951). (Structure) 

Michizo Asano and Hideo Takahashi, /. Pharm. Soc. Japan 
65 81 (1945). (Isolation from the corynebacterium ) 

874 Purpurogenone, Ci4H]oO,r,, crimson prisms, m.p. 310° (dec). 

OH OH 




Penicillium purpurogenum StoU 

Yield 8-14 g. of crude pigment from about 250 g. of dry 
mycelium, which was obtained from about 70 liters of 
culture solution. 

Ergosteryl palmitate, m.p. 104-106°, and mannitol, 
m.p. 166°, also were isolated from this fermentation. 

John C. Roberts and C. W. H. Warren, ]. Chem. Soc, 2992 
(1955). 

875 Fulvic Acid, C14H12OS, yellow crystals, m.p. 246° (dec). 

HO OH 




Penicillium flexuosum, P. brefeldianum, P. griseofulvum 

876 P. griseofulvum produced a nitrogen-containing com- 
pound, m.p. 165°, in the same broth. P. brefeldianum, 

877 produced a neutral nitrogen-containing compound, m.p. 
132-135°, in the same culture. 

Albert Edw. Oxford, Harold Raistrick and Paul Simonart, 
Biochem. J. 29 1102 (1935). (Isolation) 

F. M. Dean, R. A. Eade, R. A. Moubasher and A. Robertson, 
Nature 179 366 (1957). (Structure) 



Pfizer Handbook of Microbial Metabolites 



414 



878 Fuscin, CisHipOg, orange plates, m.p. 230°. 

O 



CH3 



.0 



HO 



-^N^N CH3 
CH3 



879 



Oidiodendron fuscum Robak 

A colorless dihydrofuscin, m.p. 206°, was also produced. 
S. E. Michael, Biochem. J. 43 528 (1948). (Isolation) 
D. H. R. Barton and J. B. Hendrickson, J. Chem. Soc, 1028 
(1956). (Synthesis) 

Azaphilones. 

This group of mold pigments, so named because most of 
them react avidly with ammonia, includes monascorubrin, 
sclerotiorin, rotiorin, rubropunctatin and monascin. 

A. Powell, A. Robertson and W. Whalley, "Chemical Society 
Symposia," Special Publication No. 5, The Chemical Society, 
London, 1956, p. 27. (Survey of the chemistry of the azaphi- 
lones to that date) 

880 Rubropunctatin, C2iHo^0-,, orange needles, m.p. 156.5° (dec), 
[a],. -3481° "(c 1.07 in chloroform). 



CH3CH2CH,CH,CH2— C=0 

I 

* CH3\ 



CH=CH— CHa 



0=Cs 



Monascus rubropunctatus Sato 

E. J. Haws, J. S. E. Holker, A. Kelly, A. D. G. Powell and 
Alexander Robertson, /. Chem. Soc, 3598 (1959). (Structure) 
A. Powell, Dissertation, Liverpool, 1954. (Isolation) 

881 Sclerotiorin, CoiHooO-.Cl, yellow crystals, m.p. 206° [a]„ +500°. 

CI CH3 CH3 



CHs— C 




CH2CH3 



415 



Pyrans and Related Substances 



Penicilliiim sclerotiorum van Beyma, P. multicolor 
G.M.P.. P. iiuplicatuui Biourgc 

Timothy P. MacCurtin and Joseph Reilly, Biochem. J. 34 
1419 (1940). (Isolation) 

H. C. Fielding, Alexander Robertson, R. B. Traners and 
W. B. WhaUey, /. Chem. Soc, 1814 (1958). 

F. M. Dean, J. Staunton and W. B. Whalley, ibid., 3004 
(1959). (Structure) 

882 Monascin, Co,Ho,;05, yellow crystals, m.p. 142°, [a]v +544°. 

Monasciis rubriginosiis Sato, M. piirpureus Wentii, 
M. rubropunctatus Sato 

Hidijiro Nishikawa, /. Agr. Chem. Soc. Japan 8 1007 
(1932). 

H. Solomon and P. Karrer, Helv. Chim. Acta 15 18 (1932). 

883 Rotiorin, C23H04O-,, red needles, m.p. 246° (dec.) (sublimes), 

[cz]d" +5080° (c 0.002 in chloroform). 
Tentative structure: 




CH0CH3 



Peiiicilliuni sclerotiorum van Beyma 

Eight kilograms of dry mycelium yielded 300-350 g. 
of sclerotiorin and 100—150 g. of rotiorin. 

G. B. Jackinan, Alexander Robertson, R. B. Traners and 
W. B. Whalley, /. Chem. Soc, 1825 (1958). (Structure) 

884 Monascorubrin, C03H26O-,, orange crystals, m.p. 134-136°, 
[alTo,/*" -1500° (c 0.1 in ethanol). 

O CH2CH2CH2CH2CH2CH2CH3 

M 




Monascus purpureus Wentii 



Pfizer Handbook of Microbial Metabolites 416 

H. Nishikawa, /. Agr. Chem. Soc. Japan 5 1007 (1932). 
(Isolation) 

K. Nakanishi, M. Ohashi, S. Kumasaki and S. Yamamura, 
J. Am. Chem. Soc. 81 6339, 6340 (1959). (Structure) 

B. C. Fielding, E. J. Haws, J. S. E. Holker, A. D. G. PoweU, 
A. Robertson, D. N. Stanway and W. B. Whalley, Tetrahedron 
Letters No. 5 24 (1960). (Proposed revised structure shown) 

885 Novobiocin (Streptonivicin, Cathomycln, Albamycin, Sphero- 
mycin, Vulcamycin, Crystallinlc Acid, Antibiotic PA-93, 
Cardelmycin), C31H36O11N2, pale yellow crystals, m.p. 
152-156° (dec.) and 174-178° (dec.) (polymorphic), 
[aJD^* -63° (c 1 in ethanol). 




HoN— C 



Streptomyces spheroides, S. niveus, S. griseus 

Herman Hoeksema, James L. Johnson and Jack W. Hin- 
man, J. Am. Chem. Soc. 77 6710 (1955). 

Jack W. Hinman, Herman Hoeksema, E. Louis Caron and 
W. G. Jackson, ibid. 78 1072 (1956). 

Clifford H. Shunk, Charles H. Stammer, Edward A. Kaczka, 
Edward Walton, Claude F. Spencer, Andrew N. Wilson, 
John W. Richter, Frederick W. HoUy and Karl Folkers, ibid. 
78 1770 (1956). (Structure) 

Herman Hoeksema, E. Louis Caron and Jack W. Hinman, 
ibid. 78 2019 (1956). (Structure) 

d. XANTHONES 
886 Ravenelin, C14H10O5, yellowish crystals, m.p. 267°. 

OH O OH 




417 Xanthones 

Helniinthosporixim ravenelii 

F. F. Nord and Robert P. Mull, Advances in Enzymol. 5 194 
(1945). (Synthesis) 

887 Rubrofusarin, CjsHjoOg, orange-red needles, m.p. 210°. 
Alternative structures : 




CH3 HO I II I CH3 



Or 




Fusarium culmorum (W.G.Sm.) Sacc, Fusarium 
graminearum Schwabe (Gibberella saubinettii) 

888 Another pigment, aurofusarin,* C30H20O10, m.p. >360° 

889 and a colorless compound, culmorin, CjgHoeOo, m.p. 175°, 
[(x]d^^ —14.45° were isolated from the same cultures. 

Julius Nicholson Ashley, Betty Constance Hobbs and Harold 
Raistrick, Biochem. J. 31 385 (1937). 

Robert P. Mull and F. F. Nord, Arch. Biochem. 4 419 (1944). 
(Structure) 

890 Asperxanthone, C^oH^^Or^, yellow needles, m.p. 203°. A 1-hy- 

droxydimethoxymethylxanthone which yields nor-rubro- 
fusarin on demethylation. 

Aspergillus niger (mycelium) 

N. A. Lund, Alexander Robertson and W. B. Whalley, 
J. Chem. Soc, 2434 (1953). 

891 Lichexanthone, CieHi405, yellowish crystals, m.p. 187°. 



CH3 O OH 



CH3O 




OCH3 



Parmelia formosana Zahlbr. 
The yield was about V2 g. from 25 g. of lichen. 
Yasuhiko Asahina and Hirashi Nogami, Bull. Chem. Soc. 
Japan 17 202 (1942). 
See entry 584. 



Pfizer Handbook of Microbial Metabolites 418 

892 Sterigmatocystin, CigHioOg, pale yellow needles, m.p. 246° 
(dec), [aln"-^ -387° (c 0.424 in chloroform). 
Probable structure: 




Aspergillus versicolor (Vuillemin) Tiraboschi 

J. E. Davies, D. Kirkaldy and John C. Roberts, J. Chem. Soc, 

2169 (1960). (Structure) 

Abou-Zeid, Dissertation, London, 1953. (Isolation) 

J. E. Davies, John C. Roberts and S. C. Wallwork, Chem. 

and Ind., 178 (1956). (Isolation) 

J. H. Birkinshaw and I. M. M. Hammady, Biochem. J. 65 

162 (1957). (Isolation) 

Yuichi Hatsuda and Shimpei Kuyama, /. Agr. Chem. Soc. 

Japan 28 989 (1954). (Chem. Abstr. 50 15,522) (Isolation) 

e. COMPOUNDS RELATED TO THIOPHENE, 
IMIDAZOLE, THIAZOLE 
AND ISOXAZOLE. 

Some of the commercially important compounds in this 
section are the antibiotics cycloserine and the penicillins 
and the vitamins, thiamine and biotin. 

Penicillin was discovered by Fleming in 1929, and com- 
mercial fermentation techniques were developed during 
the second World War. Penicillins with several different 
side-chains were found to be produced by various penicil- 
lia and aspergilli, and hundreds of unnatural penicillins 
were prepared by the addition of side-chain precursors 
to fermentations. 

It was not until 1959, however, that the nucleus com- 
mon to all penicillins, 6-aminopenicillanic acid, was iso- 
lated from fermentations.^ This discovery has made pos- 
sible the preparation of a new series of penicilUns through 

iRoichi Kato, /. Antibiotics (Japan) 6A 130, 184 (1953); F. R. 
Batchelor, F. P. Doyle, J. H. C. Nayler and G. N. Rolinson, Nature 
183 257 (1959). 



419 Thiophenes, Imidazoles, Thiazoles, Isoxazoles 

attachment of side-chains by the methods of organic 
chemistry. 

HOOC— CH N C=0 HOOC— CH N C=0 

CH3 I I I CH3 I I I O 

C CH CH— NH. C CH CH— NH— C— R 

CH3 ^ CH3 ^ 

6-Aminopenicillanic Acid Penicillins 

Since 6-aminopenicillanic acid can be isolated from 
penicillin fermentations in good yields, it is probably an 
intermediate. Also, the fact that side-chain precursors 
are so readily incorporated into the molecule indicates 
attachment of the side-chain to be the final step in peni- 
cillin biosynthesis. This is also known to be the rate-lim- 
iting step, and, even in commercial fermentations, side- 
chain precursors are added routinely. 

The precursors of the 6-aminopenicillanic acid nucleus 
have been show^n to be (stereospecifically) L-cysteine- and 
L-valine,'^ although additions of these amino acids to fer- 
mentations do not cause dramatic improvements in yields 
or in rates of synthesis. Degradation studies have shown 
that L-cysteine occurs in the same configuration after in- 
corporation into the penicillin molecule, while valine has 
been converted to the o-form. Aside from the change in 
configuration of valine, both amino acids are incorporated 
intact. 

Other substances have been considered as penicillin 
precursors and intermediates. Among them are peni- 
cillamine,^ y8-hydroxy valine,^ serine,- glycine,- homocys- 

2H. R. V. Arnsteln and P. T. Grant, Biochem. J. 57 353, 360 
(1954); H. R. V. Arnsteln and J. C. Crawhall, ibid. 67 180 (1957); 
Carl M. Stevens, Edward Inamine and Chester W. Belong, /. Biol. 
Chem. 219 405 (1956); H. R. V. Arnsteln and H. Margreiter, Bio- 
chem. J. 68 339 (1958); F. H. Grau and W. J. Halliday, ibid. 69 205 
(1957). 

^H. R. V. Arnsteln and Margaret E. Clubb, ibid. 65 618 (1957); 
Carl M. Stevens and Chester W. Belong, /. Biol. Chem. 230 991 
(1958). 

* Carl M. Stevens, Pran Vohra, Edward Inamine and Oliver A. 
Rohoh, Jr., ibid. 205 1001 (1953). 



Pfizer Handbook of Microbial Metabolites 



420 



teine,'' methionine,* glutathione* and acetate. Some of 
these rejected intermediates are shown: 

HO 









HO— C=0 


\ 


HOOC— CH— NH2 




1 


HOOC— CH— NH2 C=0 


CH3 


+ 


CH— NH2 - 


■> CH3 1 


\ 




/ 


\ CH2— CH— NH 


c 




HOCH2 


c / 


/\ 






/ \s/ 

CHs ^ 


CH3 SH 






Penicillamine 




Serine 


(3, i3-Dimethyllantliionine 


HOOC— CH— NH2 






HO 


1 




HO— C=0 


\ 


CH2 




1 


HOOC— CH— NH2 C=0 


1 


+ 


CH— NH2 - 


1 1 


CH2 




/ 


CH2 CH2— CH— NH 


\ 




HOCH2 


1 / 


SH 






CH2— S 


Homocysteine 




Serine 


Cystathionine 
HO 


HOOC— CH— NH2 




HO— C=0 


HOOC— CH— NH2 \ 


CH3 






1 


CH3 c=o 


\ 




+ 


CH— NHo - 


\ 1 


C 




/ 


C CH2— CH— NH2 


/\ 




HS— CH2 


CH3 ^ 


CH3 OH 






/3-Hydrox 


yvoline 




Cysteine 


j3, /S-Dimethyilanthionine 



Lanthionine and /?-methyllanthionine occur in several 
other polypeptide antibiotics (subtilin, duramycin, cinna- 
mycin, nisins). Certain of these compounds are incor- 
porated to some extent, but only indirectly. 

Some evidence is being accumulated concerning the 
actual peptide intermediate. The dipeptide L-cystinyl- 
L-(COOH — C^*) valine is a better penicillin precursor than 
L-(COOH — C'*) valine alone, while the reverse is true for 
protein synthesis.*' (L-Cystine can be reduced to L-cys- 
teine by the mold.) 



HOOC— CH— CH2— S 

I 
NHo 



S— CH2— CH— CO— NH— CH— COOH 

I 1 

NH2 CH 



CHs CHs 

L-CystinyI-L-(COOH— C'^) valine 
^ Carl M. Stevens, Pran Vohra, Joseph E. Moore and Chester W. 
DeLong, ibid. 210 713 (1954). 

6 H. R. V. Arnsteln and D. Morris, Biochem. ]. 71 8p (1959). 



421 Thiophenes, Imidazoles, Thiazoles, Isoxazoles 

The same research group (Arnstein and collaborators) 
has isolated a tripeptide from the mycelium of the com- 
mercial penicillin producer, Penicillium chrysogennmJ 
It is 8-(a-aminoadipyl) cysteinylvaline: 

HOOC— CH— CH.— CH.— CH2— CO— NH— CH— CO— NH— CH— COOH 

NH.> CH2SH CH 

/ \ 
CH3 CH3 

5-(a-Aminoadipyl) cysteinylvaline 

Consistent with the above evidence, this is a cysteinyl- 
valine. It is not difficult to envisage cyclization to form 
synnematin B: 

HOOC— CH NH C=0 

CH3 1 I O 

\l I II 

CH CH2— CH— NH— C— CH2— CH2— CH2— CH— COOH 

/ / I 

CH3 S NH2 

H 

., — 4H 

HOOC— CH NH C=0 

CH3 I I I O 

\l I I II 

CH CH CH— NH— C— CH2— CH2— CH2— CH— COOH 

CH3 ^ NH2 

Interesting features of this discovery are, first, that a 
side-chain is attached before cyclization to form 6-ami- 
nopenicillanic acid and, second, that the side-chain is 
a-aminoadipic acid, the side-chain of synnematin B (ceph- 
alosporin N) which is not produced by Penicillium chrys- 
ogenum. Perhaps side-chain exchange occurs after cy- 
clization. The configurations of the amino acids in the 
acyclic mycelial peptide have not been reported yet. 

The structure of cephalosporin C, a substance related 
to synnematin B, is known, but has not yet been pub- 
lished. 

Two reviews of the biosynthesis of penicillin are 
cited.s '-> 

"^ H. R. V. Arnstein, D. Morris and E. Toms, Biochim. et Biophys. 
Acta 35 561 (1959). 

® A. L. Demain, Advances in Appl. Microbiol. 1 23 (1959). 
®D. Hockenhull, Prog, in Ind. Microbiol. 1 1 (1959). 



Pfizer Handbook of Microbial Metabolites 422 

Cycloserine (oxamycin) appears to be a cyclized D-ser- 
ine amide or hydroxamide. As mentioned elsewhere it 
is known to inhibit the incorporation of D-alanyl-D-alanine 
into the cell walls of certain bacteria. 

Thiamine is an enzyme prosthetic group of fundamen- 
tal importance, probably occurring in all living things. 
Many microorganisms are capable of de novo synthesis, 
although the vitamin is required in mammalian diets. 
Some microorganisms incapable of total synthesis can 
couple certain pyrimidine and thiazole precursors, others 
require only one of the heterocycles preformed, and cer- 
tain yeasts have a requirement for thiamine itself. 

Beyond this little is known about the biosynthesis of 
thiamine. Other naturally occurring thiazoles (e.g. those 
in certain antibiotics) are known to be derivatives of 
cysteine. Nakayama has proposed the general scheme :^° 

Cysteine -^ Thiazolidine-4-carboxylic Acid — > 4-Methylthiazoie — > 

4-Methyl-5-(2-hydroxyethyl)-thiazole 

on the basis of work with mutants. Some work has been 
done on the biosynthesis of other pyrimidines, but appar- 
ently little on the thiamine constituent. 

Bacillus subtilis incorporates formate C^* extensively 
into the pyrimidine, but not the thiazole moiety of thia- 
mines^ In this bacterium the pyrimidine moiety of thia- 
mine restores growth and formate incorporation into 
purines and thymine in amethopterin treated cultures. 
The thiazole part restores thiamine synthesis, but does 
not show the additional effects. 

It appears now that all enzymes in which thiamine is 
the active site have the function of decarboxylating a-ke- 
toacids and of cleaving a-diketones or a-hydroxyketones. 
These functions were illustrated in an earlier section. 

Thiamine, unphosphorylated and detached from its 
apoenzyme, is capable of carrying out some of its coen- 
zyme functions in vitro under favorable conditions. ^-^ ^^' ^^ 

"Hideo Nakayama, Vitamins (Japan) 11 20, 169 (1956). 

SI Martin J. Pine and Robert Guthrie, J. Bacterial. 78 545 (1959). 

s- Shunzi Mizuhara and Philip Handler, /. Am. Chem. Soc. 76 571 
(1954). 

" Emeteria Yatco-Manzo, Frances Roddy, Ralph G. Yount and 
David E. Metzler, /. Biol. Chem. 234 733 (1959). 

"Ralph G. Yount and David E. Metzler, ibid. 234 738 (1959). 



423 Thiophenes, Imidazoles, Thiazoles, Isoxazoles 

By selective synthetic substitutions with blocking groups 
at various positions in the two heterocycles, the active site 
of the molecule has been located as the 2-position of the 
thiazole ring.'^' '" It is here that pyruvic acid, for ex- 
ample, is decarboxylated to form (still in combination 
with thiamine pyrophosphate) "active acetaldehyde" and 
a-ketoglutaric acid to form "active succinate." The active 
acetaldehyde intermediate was shown in Section 2. It is 
claimed that this intermediate has been isolated from 
Escherichia coli.'^'^'' 

A thorough review of thiamine is available.^" 
For more than 20 years biotin has been recognized as 
a dietary requirement in higher animals and yeasts. It 
was formerly called vitamin H, and animal deficiencies 
could be induced by feeding raw egg-white. This contains 
a protein, avidin, which complexes tightly enough with 
biotin to cause avitaminosis. 

The biochemical function and mode of action of biotin 
long remained obscure. It is now known to be a cocar- 
boxylase or coenzyme component for the transfer of car- 
bon dioxide. Some of the reactions which it catalyzes 
are: 



CH3— C— COOH ^ HOOC— CHo— C— COOH is, 19, 20, 21 
Pyruvic Acid Oxaloacetic Acid 

HOOC— CH— CHo— CH2—CH2—NH2 

I 
NHo 

Ornithine 



HOOC— CH—CH2—CH2— CHo— NH—C—NH2 22, 23 

NH2 

Citrulline 

"Ronald Breslow, /. Am. Chem. Soc. 79 1762 (1957); 80 3719 
(1958). 

^•^ Ronald Breslow and Edward McNeils, ibid. 81 3080 (1959). 

i«" Gerald L. Carlson and Gene M. Brown, /. Biol. Chem. 235 PC3 
(1960). 

^' Paul D. Boyer, Henry Lardy and Karl Myrback (Eds.), "The 
Enzymes" Academic Press, New York, 1960 Vol. II, David E. Metzler, 
Thiamine coenzymes, pp. 295-337. 

^* Henry A. Lardy, Richard L. Potter and C. A. Elvehjem, J. Biol. 
Chem. 169 541 (1947). 



Pfizer Handbook of Microbial Metabolites 424 

O O 

II II 

CH3— C=CH— C— CoA ^ HOOC— CH2— C=CH— C— CoA 24, 25 

CH3 CH3 

/3-Methylcrotonyl CoA /3-Methylglutaconyl CoA 



o o 

II II 

CH3CH2C— CoA ;:± HOOC— CH2—CH2—C— CoA 
Propionyl CoA Succinyl CoA 



26.27 



COOH 28, 29 

I 

CH2 o 

/N I H /- 

HC^ \ CH— N— C 

II CH aspartic acid | \ ^j 

C\,/ — -p >COOH C-^X 

/ V CO2 II CH 

Ribose phosphate / N 

Ribose phosphate 

(Intermediates in purine biosynthesis) 

O O 30 

II II 

CH3— C— CoA ;;::± HOOC— CH2—C— CoA 
Acetyi-CoA Malonyl-CoA 

1^ William Shive and Lorene Lane Rogers, ibid. 169 453 (1947). 

20 Herman C. Lichstein and W. W. Umbreit, ibid. 170 329 (1947). 

21 Henry A. Lardy, Richard L. Potter and R. H. Burris, ibid. 179 721 
(1949). 

22 Patricia R. MacLeod, Santiago Grisolia, Philip P. Cohen and 
Henry A. Lardy, ibid. 180 1003 (1949). 

"3 Gladys Feldott and Henry A. Lardy, ibid. 192 447 (1951 ). 

2* Bimal K. Bachhawat, Wm. G. Robinson and Minor J. Coon, 
;. Am. Chem. Soc. 76 3098 (1954); idem., J. Biol. Chem. 219 539 
(1956). 

^^ F. Lynen, J. Knappe, E. Lorch, G. Jutting and E. Ringelmann, 
Angew. Chem. 71 481 (1959). 

'*^ Henry A. Lardy and Robert Peanasky, Physiol. Rev. 33 560 
(1953). 

-' Henry A. Lardy and Julius Adler, /. Biol. Chem. 219 933 (1956). 

28 Patricia R. MacLeod and Henry A. Lardy, ibid. 179 733 (1949). 

2^ Albert G. Moat, Charles N. Wilkins, Jr. and Herman Friedman, 
ibid. 223 985 (1956); Albert G. Moat and Floyd Nasuti, Federation 
Proc. 19 313 (1960). 



425 Thiophenes, Imidazoles, Thiazoles, Isoxazoles 

The mode of action of biotin is known now in enough 
detail to suggest the scheme outUned below. It is still un- 
certain which nitrogen atom of the biotin molecule par- 
ticipates.-"^ 

O 

II 

/\ 
HN NH 

I I ATP 

HC CH > 



HoC CH—(CH..)4— COO— Enzyme 



o 

II 



o o 

T r 

Adenosine— O—P—O—P N NH 



OH OH HC CH 



CH2 CH—(CH2)4— COO— Enzyme 



or 
O O 

T T 

Adenosine — O — P — O — P O 



OH OH 



i 

O o 

II II 

©O N NH 

R I I 

R— COOH < HC CH 



HN N 

I I 

HC CH 

I I 

CH2 CH—{CH2)4— COO— Enzyme 

I CO2 



H2C CH—{CH2)4— COO— Enzyme 



^^Salih J. Wakil, Edward B. Titchener and David M. Gibson, 
Biochim. et Biophys. Acta 29 225 (1958); Salih J. Wakil, /. Am. 
Chem. Soc. 80 6465 (1958). 



Pfizer Handbook of Microbial Metabolites 426 

It is probable that biotin is attached to the enzyme in an 
amide linkage, perhaps at the t-amino group of a lysine 
unit. Evidence indicates that a variety of apoenzymes 
can use biotin as the prosthetic group in reversible carbon 
dioxide transfer just as a variety of apoenzymes can use 
riboflavin in reversible hydrogen transfer. 

Biocytin is a biotin-lysine conjugate isolated from con- 
trolled autolysates of yeast cells. ^^' ^^ 



HN NH 

1 1 

HC CH O 

1 I II 

H2C CH— (CH2)4— C— NH— (CH2)4— CH— COOH 



NH2 



Biocytin 



O 

II 

.Cx 



o 

\ 

HN NH C NH 



CH CH CH2 CH— (CH2)5— COOH 

1 I \c/ 

CH3 CH2— (CH2)4— COOH ^ 

Dethiobiotin Actithiazic Acid 

It is better utilized by some microorganisms than is biotin 
itself. 

Actithiazic acid is a biotin antimetabolite. 

The biosynthetic origin of biotin remains obscure. Pi- 
melic acid is an effective precursor in biotin-producing 
organisms. Dethiobiotin is produced by a Penicillium 
chrysogehum mutant, and it may be an intermediate in 
the biosynthetic scheme at least in this and probably in 
other microorganisms.^^ 

^^ Lemuel D. Wright, Emlen L. Cresson, Helen R. Skeggs, Thomas 
R. Wood, Robert L. Peck, Donald E. Wolf and Karl Folkers, ibid. 74 
1996 (1952). 

■^- Donald E. Wolf, John Valiant, Robert L. Peck and Karl Folkers, 
ibid. 74 2002 (1952). 

3'' E. L. Tatum, /. Biol. Chem. 160 455 (1945). 



427 Thiophenes, Imidazoles, Thiazoles, Isoxazoles 

Junipal appears to be related to the acetylenic* sub- 
stances typical of basidiomycetes which were listed in an 
earlier section. In some way sulfur seems to have been 
added, in effect across two acetylenic bonds to form a 
thiophene ring. It has been suggested" that junipal and 
anisaldehyde, occurring in the same culture and with the 
same number of carbon atoms, may be derivatives of a 
common acetylenic aldehyde precursor, perhaps C7H4O: 

CHO 
HC = C— C = C— CH=CH— CHO 



CHO 




OCH 



Azomycin seems to incorporate a modified guanidine 
group. 

893 Azomycin (2-Nitroimidazole), C3H3O2N3, white needles, m.p. 
283° (dec). 



N NH 

I 

N02 

Nocardia sp. resembling N. mesenterica 
Shoshlro Nakamura and Hamao Umezawa, /. Antibiotics 
(Japan) 8A 66 (1955) and other papers in this series. 

894 Oxamycin (Cycloserine, Orientomycin D-4-Amino-3-isoxazoli- 
done, PA-94), CsHgOoNo, colorless crystals, m.p. 156° 
(dec), [a]546i 25° +137° ±2° (c 5 in 2 N sodium hydrox- 
ide). 

CH2 CH— NH2 

I I 

o c=o 

H 

Streptomyces garyphalus, S. orchidaceus, S. lavendulae, 
S. nagasakiensie nov. sp., S. K-300, etc. 

^-i J. H. Birkinshaw and P. Chaplen, Biochem. J. 60 255 (1955). 



Pfizer Handbook of Microbial Metabolites 428 

Dale A. Harris, Myrle Ruger, Mary Ann Reagan, Frank J. 
Wolf, Robert L. Peck, Hyman Wallick and H. Boyd Woodruff, 
Antibiotics and Chemotherapy 5 183 (1955). 

Roger L. Harned, Phil Harter Hidy and Eleanore Kropp 
LaBaw, ibid. 5 204 (1955). 

Charles H. Stammer, Andrew N. Wilson, Claude F. Spencer, 
Frank W. Bachelor, Frederick W. Holly and Karl Folkers, J. 
Am. Chem. Soc. 79 3236 (1957). (Synthesis) 

895 Junipal, CgHfiOS, thick, colorless needles, m.p. 80°. 

HC CH 

II II 

CH3— C = C— C C— CHO 

Daedalea juniperina Murr. 

J. H. Birkinshaw and P. Chaplen, Biochem. J. 60 255 
(1955). 



896 6-Methoxybenzoxazolidone, CgHyOgN, red crystals, m.p. 154°. 

V 



CH3O o 




c=o 



Ustilago maydis (spores) 

The same compound has been isolated from young com 
plants. 

P. H. List, Arch. Pharm. 292 452 (1959). 

897 6-Aminopenicillanic Acid, CgHigOgNsS, colorless crystals, m.p. 
208° (dec). 

HOOC— CH N C=0 

CH3 1 



CH3 



C CH CH— NH2 



Penicillium chrysogenum 

F. R. Batchelor, F. P. Doyle, J. H. C. Nayler and G. N. Robin- 
son, Nature 183 257 (1959). 



429 



Thiophenes, Imidazoles, Thiazoles, Isoxazoles 



898 5-Amino-4-imidazolecarboxamide Riboside, CoHi40r,N4, colorless 
crystals, m.p. 213° (dec, previous browning). 



II 



HOCH 




OH OH 

Escherichia coli (sulfonamide— inhibited) 
G. Robert Greenberg and Edra L. Spilman, /. Biol. Chem. 
219 411 (1956). 

899 Actithiazic Acid (Acidomycin, Mycobacidin PA-95), CyHigOgNS, 
colorless needles, m.p. 140°, [aJD'^ —60° (c 1 in absolute 
alcohol). 



o=c- 



-NH 



CH2 



CH— CH2CH2CH2CH2CH2COOH 



Streptomyces virginiae, S. cinnamonensis, S. lavendu- 
lae 

Yields of about 0.3 g. per liter have been reported. 

Walton E. Grundy, Alma L. Whitman, Elbina G. Rdzok, 
Edward J. Rdzok, Marjorie E. Hanes and John C. Sylvester, 
Antibiotics and Chemotherapy 2 399 (1952). 

J. R. Schenck and A. F. DeRose, Arch. Biochem. and 
Biophys. 40 263 (1952). 

R. K. Clark, Jr. and J. R. Schenck, ibid. 40 270 (1952). 

W. M. McLamore, Walter D. Celmer, Virgil V. Bogert, Frank 
C. Pennington and I. A. Solomons, /. Am. Chem. Soc. 74 2946 
(1952). 

B. A. Sobin, ibid. 74 2947 (1952). 

W. M. McLamore, Walter D. Celmer, Virgil V. Bogert, Frank 
C. Pennington, B. A. Sobin and I. A. Solomons, ibid. 75 105 
(1953). (Synthesis) 



Pfizer Handbook of Microbial Metabolites 



430 



900 Biotin, CjoHifiOaNoS, colorless needles, m.p. 230-232° (dec), 
[ale" +92° (c 0.3 in 0.1 N sodium hydroxide). 



HN 



HC- 



NH 



-CH 



CH2 CH— CHCHiCHoCH-COOH 

Torula utilis, other yeasts (occurs also in molds and 
bacteria) 

Yields of 0.5-3.6 /xg. per gram of dry cell weight are ob- 
tained from Torula utilis. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Pubhshing Co., Inc., New York, 
1954 Vol. II, J. M. Van Lanen, Production of vitamins other 
than riboflavin, chap. 6, pp. 191-216. (A review) 

901 Biotin- 1 -sulfoxide, Ci(,Hic04N2S, colorless crystals, m.p. 238- 
243°, [a] I,-" -40° (in 0.1 N sodium hydroxide). 



HN NH 



CH CH 



CH2 CH— CH2CH2CH2CH2COOH 



Aspergillus iiiger 

Lemuel D. Wright and Emlen L. Cresson, /. Am. Chem. Soc. 
76 4156 (1954). 

Lemuel D. Wright, Emlen L. Cresson, John Valiant, Don- 
ald E. Wolf and Karl Folkers, ibid. 76 4160, 4163 (1954). 
(Isolation and characterization) 



431 Thiophenes, Imidazoles, Thiazoles, Isoxazoles 

902 Dethiobiotin (Desthiobiotin, 5-Methyl-2-oxo-4-imidazolidineca- 
proic Acid), C,,,H,,sO;(No, colorless needles, m.p. 156-158°, 
[all,-' +10.7° (c 2.0 in water). 



HN NH 




CHs (CHolsCOOH 

Penicillium chrysogenum 

E. L. Tatum, 7. Biol. Chem. 160 455 (1945). 

903 Thiamin (Vitamin B,, Aneurin) (Chloride Hydrochloride), 

Ci.HjsOi\4CLS, colorless needles, m.p. ~250° (dec). 

NHoHCI CHs CHoCHoOH 

CHo \ / 

CI® 

Most yeasts, molds and bacteria 

Yields of 120-200 /xg. per gram of dry primary-grown 
yeast cells can be obtained. Much higher yields (600- 
1200 /xg. per gram) can be obtained if all that is required 
is coupling of supplied precursors. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical PubHshing Co., Inc., New York, 
1954 Vol. II, J. M. Van Lanen Production of vitamins other 
than riboflavin, chap. 6, pp. 191-216. (A review) 

904 Cocarboxylase (Cozymase II, Vitamin Bi-diphosphate, Thiamin 

diphosphate, Aneurindiphosphate) (Hydrochloride), 
Ci^HisO-N^SP.-HCl, nearly colorless needles, m.p. 242- 
244° (dec). 




C— CHs O O 




CH C— CH2— CH2— O— P— O— P=0 

I I 

OH OH 



Pfizer Handbook of Microbial Metabolites 432 

Yeast 

K. Lohmann and Ph. Schuster, Biochem. Z. 294 188 (1937). 
(Isolation) 

Kurt G. Stern and Jesse W. Hofer, Science 85 483 (1937). 
(Synthesis) 

905 Synnematin-B (Cephalosporin N, Salmotin), C14H21O6N3S, bar- 

ium salt, [xW° +187° (c 0.6 in water). 

HOOC— CH N— C=0 

CH3 
\ 

C CH— CH— NH— CO— CH2— CH2— CH2— CH— COOH 

/ \c/ 1 

CHs ^ NH2 

Cephalosporium salmosynnematum 

E. P. Abraham, "CIBA Lectures in Microbiol. Biochemistry," 
Biochemistry of some peptide and steroid antibiotics, John 
Wiley and Sons, New York, 1957. (A review) 

Natural Penicillins. General formula: 

HOOC— CH N C=0 

CH3 I II O 

C CH CH— NH— C— R 

CHs ^ 

906 Penicillin G, Ci„Hi804N2S, colorless prisms, m.p. (Na salt) 215° 

(dec), [aW'-' +305° (c 0.821 in water). 

R = Benzyl <f \-CH2 



907 Penicillin K, C16H26O4N2S, colorless prisms (Na salt), [aln"'^ 

+258° (c 0.43 in water). 

R = n-Heptyl CHslCHzle" 

908 Penicillin X, CkjHisOjNsS, colorless crystals, m.p. (Na salt) 

228-235° (dec), [a]v +267° (c 0.525 in water). 

R = p-Hydroxybenzyl HO— f y-CH2- 



909 Gigantic Acid (Dihydro F), C14H20O4N2S (Na salt), colorless 
crystals, m.p. 188° (dec), [czId'' +319° (c 1 in water). 

R = n-Amyl CH3(CH2)4~ 



433 Thiophenes, Imidazoles, Thiazoles, Isoxazoles 

910 Penicillin F (Flavicidin, Flavicin) Ci4H.o04N.,S, m.p. (Na salt) 

204° (dec), [oc]v-"-' +276-316° (c 0.821 in water). 

R = n-Pentenyl CH3CH,— CH=CH— CH2— 

The A"-pentenyl variant is also known. 

Penicillium species, especially P. chrysogenum and 
P. notatum Westling, and aspergillus species, especially 
A. fiaviis from which Penicillin F was obtained. 

H. Clarke, J. Johnson and R. Robinson, "The Chemistry of 
Penicilhn," Princeton University Press, Princeton, 1949. (A 
review ) 

911 Cephalosporin C, proposed molecular formula CigHsiOgNgS, Na 

salt: [aW +103°. 

Structural features: 

Acid hydrolysis yields 1 D-a-aminoadipic acid, 1 CO2 
and 2 NH3. No penicillamine is produced in contrast to 
cephalosporin N. 

Cephalosporium salmosynnematum 

G. G. F. Newton and E. P. Abraham, Biochem. J. 62 651 
(1956). 

912 Biocytin, C16H08O4N4S, colorless crystals, m.p. 228-230° (dec.) 

(245-252°). 

O 

/K 

HN NH 

I I 

CH CH 

I I 

CH2 CH— (CH2)4— CO— NH— (CH2)4— CH— COOH 

^ NH2 

Yeast 

Lemuel D. Wright, Emlen J. Cresson, Helen R. Skeggs, 
Thomas R. Wood, Robert L. Peck, Donald E. Wolf and Karl 
Folkers, /. Am. Chem. Soc. 72 1048 (1950). (Isolation) 

Robert L. Peck, Donald E. Wolf and Karl Folkers, ibid. 74 
1999 (1952). (Structure) 

Donald E. Wolf, John Valiant, Robert L. Peck and Karl 
Folkers, ibid. 74 1002 (1952). (Synthesis) 



Pfizer Handbook of Microbial Metabolites 



434 



f. PYRROLES, PORPHYRINS AND RELATED COMPOUNDS 

Pyrroles occur rather frequently as microorganism me- 
tabolites. They are constituents of porphyrins, of vitamin 
B^o, of certain bacterial pigments, and of some compounds 
which have been considered as antibiotics. 

More has been published concerning the biosynthesis 
of the complex substances because of their more general 
import in biological systems, but it is tempting to specu- 
late on the origins of the simpler compounds even though 
little evidence is yet available. 

Holomycin is the simplest of three similar substances 
produced by streptomycetes, although the structures of 
aureothricin and thiolutin were determined earlier. The 
skeletons of glycine and cysteine are perceptible within 
the holomycin molecule, and, superficially, it seems that 
a biosynthetic route related to the following might take 
place : 

NHo 



HOOC— CH— CH2 

1 II 

S C— OH 

/ 1 
S CH 

^"2 NHo 


~ 




CH2— NHolCOCHs) 

c=o 

/ 
HO 


— H2O 




Cystine 


Glycine 




NH2 






HOOC— CH— CHo 








S C— OH CH,— NHzlCOCHs 



_ S CH C=0 

XH2^ ^NH^ 
I 
NH2 

I 
HOOC— CH— CH2 O 

I II 

S C- CH— NH2(COCH3 



— H2O 



/ 



\CH2^ \n/ 



CH C=0 
H 



II 



435 Pyrroles, Porphyrins and Related Compounds 

NH.. 
HOOC— CH— CH, O 

S C CH -NHo(COCH3) 

S C CO 

H 

III 

/\ 

S C CH— NH—CO—CH:, 

111 
CH C C=0 + Serine or Alanine 

\ / 

H 

Holomycin 

A glycylcystine intermediate I is reminiscent of the pep- 
tide intermediate now implicated in the biosynthesis of 
penicillin.^ It is known that there has been some aca- 
demic interest in the origin of these compounds, however, 
and since no publications have been forthcoming, perhaps 
the problem is more complicated. 

Pyoluteorin, with a carbonyl group at the two position 
of the pyrrole moiety, suggests an origin in the glutamate 
-^ prohne pathway, perhaps from 8^-pyrroline-5-carbox- 
ylic acid, although the chlorination of the ring may indi- 
cate a less obvious derivation. The pyrrohdine moiety of 
the plant alkaloid, nicotine, has been shown to be bio- 
synthesized from glutamate.- 

The origins of prodigiosin and netropsin are not obvious. 
Some work has been done on prodigiosin.^ * Glycine-2-C^* 
was incorporated into prodigiosin, but 5-aminolevulinic 
acid-5-C^* was not.* This apparently distinguishes 
the method of biosynthesis from that of the por- 
phyrins. Moreover, C^ '-labeled L-proline was found to be 
several times more efficient as a prodigiosin precursor in 
Serratia marcescens than glycine, while the reverse is 

^ H. R. V. Arnstein, D. Morris and E. Toms, Biochim. et Biophys. 
Acta 35 561 (1959). 

- Thomas Griffith, Kenneth P. Hellman and Richard U. Byerrum, 
J. Biol. Chem. 235 800 (1960). 

3R. Hubbard and C. Rimlngton, Biochem. J. 46 220 (1950). 

^ Gerald S. Marks and Lawrence Bogorad, Proc. Nat. Acad. Sci. 46 
25 (1960). 



Pfizer Handbook of Microbial Metabolites 436 

true in heme synthesis (in rats)."* The biosynthesis at 
least seems to be related to the metabolism of 5-carbon 
units such as proUne, ornithine and glutamic acid. It 
was further proposed^ that the methoxyl group in one pyr- 
role ring indicated derivation from hydroxyproHne, and 
that the colorless Cjo pyrrolic substance, which is thought 
to be a prodigiosin precursor/' was also probably derived 
from two C-5 units and that the w-amyl side-chain also 
might be a rudimentary C-5 amino acid chain. In this 
connection, the isolation of a C25 "prodigiosin-like pig- 
ment"' from a streptomycete should be mentioned. While 
all of the proposals made are not entirely compatible with 
the revised structure published since,^ the basic tenets 
seem to be sound. 

Orange and blue variants of prodigiosin occur. The 
latter, which are less soluble, may be metal chelates. 

Some work also has been done on the biosynthesis of 
the pyrrolic pigments of Bacillus bruntzii, and glycylgly- 
cine was found to be a better precursor than glycine and 
a number of other peptides.^ 

It is safe to say that natural pyrroles are formed by a 
variety of methods. Demonstration of the participation 
of erythrose in the shikimic acid biosynthetic route has 
inspired the admonition that erythrose and its 4-C-atom 
derivatives should not be ignored as possible precursors 
of furans and pyrroles.^" 

Because of their importance in photosynthesis, in he- 
moglobin, in cytochromes and peroxidases and in the 
chromophore of vitamin B12, there has been much investi- 
gation of the general mode of biosynthesis of porphyrins. 
It is likely that a similar method is operative in all cases. 

Porphyrins are present in yeasts, molds and bacteria. 

= David Sherain and D. Rittenberg, J. Biol. Chem. 166 621 (1946). 

^ Ursula V. Santer and Henry J. Vogel, Biochim. et Biophys. Acta 
19 578 (1956). 

^ F. Arcamone, A. DiMarco, M. Ghione and T. Scottl, Giom. 
microbiol. 4 77 (1957). 

^ Harry H. Wasserman, James A. McKeon, Lewis Smith and Peter 
Forgione, J. Am. Chem. Soc. 82 506 (1960). 

^ J. G. Marchal and S. Baldo, Trav. lab. microbiol. fac. pharm. 
(Nancy) No. 18 187 (1956). 

'"Ernest Wenkert, Experientia 15 166 (1959). 



437 



Pyrroles, Porphyrins and Related Compounds 



The photosynthetic bacteria, grown aerobically in light, 
are a rich source, and so are corynebacteria. Part of the 
biosynthetic pathway to the porphyrins has been explored 
in photosynthetic bacteria, and it is thought to be of gen- 
eral significance:^^' ^- 



-CO. 



HOOC— CH>— CH>— C— COOH 
a-Ketoglutaric Acid 



-^ HOOC— CH.— CHo— CO— CoA 
Succinyl Coenzyme A 



HOOC— CHo— CHo— CO— CoA + HOOC— CH2 
Succinyl Coenzyme A Glycine 



-NH, 



-COo 



COOH 



HOOC— CH2— CHo— C— CHo— NH2 
6-Aminolevulinic Acid 



H2N— CH2 



COOH CH2 

1 1 
CH2 CH2 

1 1 


— 2H20 






> 




CH2 o=c 

1 1 








C CHo 
H2N 








5-Aminolevulinic Acid 








HOOC— CH2 

\ 

c- 

c 

H2N— CH2/ \ 


CH2— CH2- 
/ 

— c 

II 

c 


-COOH 






H 






Porphobilinogen 





Pyridoxal phosphate is required as a co-factor (glycine 
activator) in the glycine-succinyl-COA condensation.^^ 
Porphobihnogen then condenses to form coproporphyrin 
and protoporphyrin. In certain photosynthetic bacteria, 

" June Lascelles, Biochem. }. 62 78 (1956); idem., Abstracts of the 
Gordon Conference on Metabolism, 1957. 

^- Goro Kikuchi, Abhaya Kumar, Phyllis Talmadge and David 
Shemin, /. Biol. Chem. 233 1214 (1958). 



Pfizer Handbook of Microbial Metabolites 43S 

such as Rhodopseudomonas spheroides, the following se- 
quence has been shown: 

— 4NH3 — 4H 

4 Porphobilinogen > Uroporphyrinogen > Uroporphyrin III 

— 2H 

— 4CO2 

— 4H 
Coproporphyrinogen > Coproporphyrin III 

--4H — 2CO2 

— 4H 
Protoporphyrinogen > Protoporphyrin IX 

The reduced precursors may be the biologically active 
species, and the porphyrins by-products stabilized by oxi- 
dation.'^ 

Higher animals (as well as microorganisms) are ca- 
pable of porphyrin synthesis, and, in fact, the above work 
with photosynthetic bacteria was based on earUer labeling 
experiments in animals,^' and porphobilinogen was first 
isolated from the urine of humans with acute porphyria." 

Widely occurring enzymes convert porphobilinogen to 
uroporphyrins, but it is difficult to isolate and identify the 
intermediates. Apparently they are quite transitory. 
Some interesting speculations have been published con- 
cerning their nature.'^' ^''' The Wittenberg hypothesis, 
based on the known transformations of porphobilinogen 
by chemicals and enzymes, the extensive labeling studies 
that have been published, and on the construction of mod- 
els, is outlined in the following series of equations : 

'•"'David Shemin and D. Rittenberg, J. Biol. Chem. 166 621, 627 
(1946); Norman S. Radin, D. Rittenberg and David Shemin, ibid. 184 
745 (1950); Jonathan Wittenberg and David Shemin, ibid. 185 103 
(1950); David Shemin and Jonathan Wittenberg, ibid. 192 315 
(1951); Helen M. Muir and A. Neuberger, Biochem. J. 47 97 (1950); 
David Shemin^ Charlotte S. Russell and Tessa Abramsky, /. Biol. 
Chem. 215 613 (1954); K. D. Gibson, W. G. Lauer and A. Neuberger, 
Biochevi. J. 70 71 (1958); K. D. Gibson, A. Neuberger and J. J. Scott, 
ibid. 61 618 (1955); J. E. Falk, E. I. B. Dresel, A. Benson and B. C. 
Knight, ibid. 63 87 (1956); E. I. B. Dresel and J. E. Falk, ibid. 63 
388 (1956). 

1" R. G. Westall, Nature 170 614 (1952); G. H. Cookson and C. Rim- 
ington, Biochem. J. 57 476 (1954). 

'^ David Shemin, Harvey Lectures 50 258 (1956). 

16 Jonathan B. Wittenberg, Nature 184 876 (1959). 



439 



Pyrroles, Porphyrins and Related Compounds 



A = -CHuCOOH 
P = -CH;CH:COOH 




A 


P / 
H, 


V 








/\N/ 




\n/ 


H 




H 


H,C „ 






\ H 


1 C r 


/Kv 


2 




H, 

J 





Bogorad found'' that the enzyme porphobilinogen de- 
aminase converts porphobilinogen (A) to uroporphyrino- 
gen (D). Because a second enzyme, uroporphyrinogen 
isomerase, has as its only substrate (not D) a product of 
the action of porphobilinogen deaminase on (A), there 
must have been one or more colorless intermediates. The 
intermediates must be convertible, spontaneously or under 
the continuing influence of porphobihnogen deaminase. 



1' Lawrence Bogorad, /. Biol. Chem. 233 501, 510, 516 (1958). 



Pfizer Handbook of Microbial Metabolites 440 

to (D) (reaction 5). The linear tetrapyrrole (B) shown is 
the intermediate proposed by Wittenberg. 

The enzyme, uroporphyrinogen isomerase, acting on 
porphobihnogen, yields uroporphyrinogen III (E) as its 
first detectable product. Wittenberg proposed that the 
function of this enzyme is to condense 2 molecules of (B) 
(reaction 2), creating the cychc octapyrrole (C). Model 
studies indicate that such an intermediate could fold and 
undergo rearrangement, spontaneously or under contin- 
ued enzyme influence, to yield 2 molecules of uropor- 
phyrinogen III (E) (reaction 4). 

The over-all result of this reaction sequence would be 
the interchange of the pyrrole moieties destined to form 
rings D of the porphyrins between two tetrapyrroles, with 
consequent reversal of the positions of the D rings rela- 
tive to the other pyrrole rings of the tetrapyrroles. 

This hypothesis seems to be in accord with aU other 
known evidence concerning porphyrin biosynthesis, and 
it would account for their pecuhar asymmetry. Many ref- 
erences to related work are cited by Wittenberg. It is 
notable that appropriate dipyrrome thanes were not con- 
verted to porphyrinogens or porphyrins by porphobilino- 
gen deaminase. ^^ 

Vitamin 6^2 is the only vitamin produced exclusively by 
microorganisms, although not all microbes are capable 
of elaborating it. Most seem to form little more than 
enough for their own slight requirements, the best or- 
ganisms for primary production by fermentation being: 
Streptomyces olivaceus, S. griseus, Propionibacterium 
shernianii and Bacillus megatherium. 

The nucleus of vitamin Bj^ differs somewhat from that 
of porphyrins and is called the corrin ring:^"' 




17 -15. 13 
Corrin 

18 D. S. Hoare and H. Heath, Biochim. et Biophys. Acta 39 167 
(1960). 



441 



Pyrroles, Porphyrins and Related Compounds 



(b) COoH 



CH2 CH3 \ XHi.CO^H (c) 




CH2.CH2.CO2H (d) 



(g) HO2C.CH2 



(f) H02C.CH2.CK CH3 '-"3 



CH2.CH2.CO2H (e) 
Cobyrinic Acid 



CONH 
/ 
CONH2CH3 CH2 u 

/ H V 
CH^ *?">N^ CN/^-pl^ 



CH3 








NH2 

-CO 

/ AM 

NH OH 

/ O©/ 

CH2 / / 

a-(5,6-Dimethylbenzimidazolyl) cobamide Cyanide 



The nucleotide-free carboxylic acid form is called co- 
byrinic acid, the carboxyl groups (amides, etc.) being let- 



Pfizer Handbook of Microbial Metabolites 



442 



tered as shown. When the aminopropanol group is in- 
corporated by amide Unkage at the f-position, the name is 
modified to cobinic acid ( all carboxyl groups as amides = 
cobinamide), and when ribose is incorporated, the name 
is modified to cobamic acid (cobamide). The name of 
the heterocycle is then inserted at the beginning with the 
suffix -yl. Thus, vitamin B12 is correctly named: a-(5,6- 
dimethylbenzimidazolyl) cobamide cyanide. 

The two principal moieties are called the planar group 
and the nucleotide, and these are essentially perpendicu- 
lar in relative steric arrangement. 

A number of analogues of vitamin Bjo have been iso- 
lated from natural sources. These sources include B12 
fermentations, the rumen or the gut of various animals 
and sewage sludge. The naturally occurring analogues 
are listed below by trivial name, together with the charac- 
teristic heterocycle of the nucleotide. 



TABLE I 
Naturally Occurring Vifamin 612 Analogues 



Name 


Nucleotide base 


Reference 


Vitamin Bn 


5,6-Dimethylbenzimidazole 






Pseudo (i/')-Vitamin B12 


Adenine 


19, 20, 21, 


22,23 


Factor A 


2-Methyladenine 


20, 21, 22, 


23 


Factor B (Etiocobalamine) 


No nucleotide 


21, 24, 25, 


26 


Factor C (Guanosine Diphos- 








phate Factor B) 


Guanine 


21, 24, 25, 


27,37 


Factor D* 


Unknown 


21,22 




Factor E* 


Unknown 


22 




Factor F 


2-Methylmercaptoadenine(?) 


21,22,25 




Factor G 


Hypoxanthine 


22 




Factor H 


2-Methylhypoxanthine 


22 




Factor 1 (B12 factorni) 


5-Hydroxybenzimidazole 


22, 28, 29 




Factors J, K, L, M 


Unknown 


30 






Unknown purine base (?) 
2-Methylmercaptoadenine 


31 
32 




(May be factor F) 




(May be factor C) 


Guanine 


27 




Factor "A" Ribose Phosphate 


No base 


33 




Factor Via (Cobyrinic Acid 








a, b, c, d, e, g-hexaamide) 


No base 


34,35 




Factor Vib (Cobyrinic Acid 








Pentamide) 


No base 


34, 35 






5-Methylbenzimidazole 


36 










Benzimidazole 


36t 









*Not crysfoillne. t About 17 other cobamides were defected in this study. 



443 Pyrroles, Porphyrins and Related Compounds 

Besides the natural analogues, many vitamin 8,0 "var- 
iants have been prepared by addition of analogues of the 

1-' H. W. Dion, D. G. Calkins and J. J. Pfiffner, /. Am. Chem. Soc. 74 
1108 (1952). 

-"]. E. Ford, E. S. Holdsworth, S. K. Kon and J. W. G. Porter, 
Nature 171 148 (1953). 

-^ H. G. Heinrich (Editor), "Vitamin Bu and Intrinsic Factor, First 
European Symposium, Hamburg, 1956." M. E. Coates and S. K. Kon, 
Ferdinand Enke. Stuttgart, 1956, p. 72. 

-- F. B. Brown, J. C. Cain, Dorothy E. Gant, T. F. J. Parker and 
E. Lester Smith, Biochem. J. 59 82 (1955). 

-' H. W. Dion, D. G. Calkins, and J. J. Pfiffner, /. Am. Chem. Soc. 
76 948 (1954). 

^S. K. Kon, Biochem. Symposium No. 13, p. 17 (1955). 

^'' H. G. Heinrich (Editor), "Vitamin B,o and Intrinsic Factor, First 
European, Hamburg, 1956." J. W. S. Porter, Ferdinand Enke, 
Stuttgart, 1956, p. 43. 

-•'J. B. Armitage, J. R. Cannon, A. W. Johnson, T. F. J. Parker, 
E. Lester Smith, W. H. Stafford and A. R. Todd, /. Chem. Soc, 3849 
(1953). 

-" R. Barchielli, G. Boretti, P. Julita, A. Migliacci and A. Minghetti, 
Biochim. et Biophijs. Acta 25 452 (1957). 

-* Wilhelm Friedrich and Konrad Bernhauer, Chem. Ber. 89 2030 
(1956). 

-^ Wilhelm Friedrich and Konrad Bernhauer, Angew. Chem. 65 627 
(1953). 

'" Clifford H. Shunk, Franklin M. Robinson, James F. McPherson, 
Marjorie M. Gasser and Karl Folkers, 7. Am. Chem. Soc. 78 3228 
(1956). 

31 G. E. W. Wolstenholm and Maeve O'Connor (Eds.), CIBA 
Foundation Symposium on "The Chemistry and Biology of Purines," 
E. Lester Smith, The chemistry of neiv purines in the Bi2 series of 
vitamins, Little, Brown & Co., Boston, 1957, pp. 160-168. 

3- Wilhelm Friedrich and Konrad Bernhauer, Chem. Ber. 90 1966 
(1957). 

■'^ Hanswerner Dellweg and Konrad Bernhauer, Arch. Biochem. and 
Biophijs. 69 74 (1957). 

■''* Konrad Bernhauer, Hanswerner Dellweg, Wilhelm Friedrich, 
Gisela Gross and F. Wagner, Z. Naturforsch. 156 336 (1960). 

3^" K. Bernhauer, H. W. Dellweg, W. Friedrich, G. Gross, F. Wagner, 
and P. Zeller, Helv. Chim. Acta 43 693 (1960). 

^^ Konrad Bernhauer, Elisabeth Becher, Gisela Gross and Georg 
Wilharm, Biochem. Z. 332 562 (1960). 

3^" K. Bernhauer, F. Wagner and P. Zeller, Helv. Chim. Acta 43 696 
(1960). 

""Wilhelm Friedrich and Konrad Bernhauer, Chem. Ber. 91 2061 
(1958). 

^" G. Boretti, A. DiMarco, T. Fuoco, M. P. Marnati, A. Migliacci and 
C. Spalla, Biochim. et Biophijs. Acta 37 379 (1960). 



Pfizer Handbook of Microbial Metabolites 444 

nucleotide base to fermentations. A review^^ lists about 
50 such compounds, some of which have vitamin activity. 

There seems to be a fundamental similarity in the bio- 
synthetic routes to vitamin B^o and the porphyrins. C^^- 
Labeled glycine or 8-aminolevuhnic acid are heavily in- 
corporated.-^'* Threonine furnishes the aminopropanol 
moiety as demonstrated by incorporation of the amino 
acid labeled with N^^^" There seems to be no information 
yet on the biosynthetic origin of the dimethylbenzimida- 
zole moiety. 

Red cobamide-containing polypeptides have been iso- 
lated from microorganisms, and some of these can replace 
cobamide in deficient microorganisms, and in the oral 
treatment of pernicious anemia.*^' ^- 

Cobamides have been impUcated in several metabolic 
processes.*^ In Escherichia coli mutants they seem to 
assist in the formation and transfer of methionine methyl 
groups" (folic acid is also required). They are thought 
to be involved in the reduction of disulfides to thiols.*^ In 

38 D. Perlman, Advances in Appl. Microbiol. 1 87-112 (1952). (A 
review) 

2^ David Shemin, John W. Corcoran, Charles Rosenblum and Ian 
M. Miller, Science 124 272 (1956); John W. Corcoran and David 
Shemin, Biochim. et Biophys. Acta 25 661 (1957). 

*° Alvin I. Krasna, Charles Rosenblum and David B. Sprinson, /. 
Biol. Chem. 225 745 (1957). 

" H. G. Wijmenga, J. Lens and A. Middlebeek, Chem. Weekblad 45 
342 (1949); H. G. Wijmenga and B. Hurenkamp, ibid. 47 217 (1951); 
H. G. Wijmenga and W. L. C. Veer, ibid. 48 33 (1952); H. G. Wij- 
menga, K. W. Thompson, K. G. Stern and D. J. O'Connell, Biochim. 
et Biophys. Acta 13 144 (1954); H. G. Wijmenga, J. Lens and S. J. 
Geerts, Acta Haematol. 11 372 (1954). 

i^K. Hausmann, Lancet 257 962 (1949); K. Hausmann and K. 
MulU, Acta Haemotol. 1 345 (1952); idem.. Lancet 262 185 (1952); 
K. Hausmann, Klin. Wochschr. 31 1017 (1953); K. Hausmann, L. 
Ludwig and K. Mulli, Acta Haemotol. 10 282 (1953); K. Mulli and 
O. J. Schmid, Z. Vitamin-, Hormon-u. Fermentforsch. 8 225 (1956); 
J. G. Heathcote and F. S. Mooney, Lancet 274 982 (1958). 

43R. D. Wilhams (Ed.), "The Biochemistry of Vitamin B,..," June 
Lascelles and M. J. Cross, The function of vitamin B,j in microorgan- 
isms. Biochemical Society Symposia No. 13, Cambridge University 
Press, London, 1955, pp. 109-123. 

•i^C. W. Helleiner and D. D. Woods, Biochem. J. 63 26p (1956). 

■•^ J. W. Dubnoff and E. Bartroy, Arch. Biochem. and Biophys. 62 
86 (1956); Chiun T. Ling and Bacon F. Chow, J. Biol. Chem. 206 797 
(1954). 



445 Pyrroles, Porphyrins and Related Com|>ounds 

Lactobacillus leichmannii they are required for the re- 
duction of formate to the methyl group of thymine by a 
pathway not involving methionine nor a hydroxymethyl 
intermediate.^" In the same organism they have been 
reported necessary for the synthesis of deoxyribose."*^ 

The isolation of actual coenzyme forms of cobamides 
has permitted more precise determination of some func- 
tions which are known to be direct. Barker and collab- 
orators found that cell-free extracts of the anaerobe Clos- 
tridium tetavomorphuin metabolized glutamate in a way 
different from the citric acid cycle, catalyzing the equi- 
librium : 

HOOC— CHo— CHo— CH— COOH ;=i HOOC— CH— CH— COOH 

I I I 

NH2 CH3 NH2 

Glutamic Acid /3-MethyIaspartic Acid 

HOOC— CH 

CH3— C— COOH 

Mesaconic Acid 

An orange form of pseudovitamin B12 was isolated and 
found to be required for the first step.*® (It is noteworthy 
that /jj-methylaspartic acid occurs in the polypeptide anti- 
biotic, aspartocin.) The entire nature of this coenzyme 
is still unknown, but the nucleotide base is known to be 
adenine. Also a second mole of adenine nucleoside is 
present, bound in such a way as to affect radically the 
corphyrin spectrum, and cleavable by photolysis. The 
nucleoside apparently is attached to cobalt, replacing the 
cyano group. It contains an unusual sugar.*'' 

In this isomerization there are two possible migrating 

**' James L. Dinning, Barbara K. Allen, Ruth Young and Paul L. 
Day, J. Biol. Chem. 233 674 (1958). 

■•" Mancourt Downing and B. S. Schwelgert, /. Biol. Chem. 220 521 
(1956); W. T. Wong and B. S. Schwelgert, Proc. Soc. Exptl. Biol. 
Med. 94 455 (1957). 

*s H. A. Barker, H. Welssbach and R. D. Smyth, Proc. Nat. Acad. 
Sci. U. S. 44 1093 (1958). 

■*9 H. A. Barker, R. D. Smyth, H. Weissbach, J. I. Toohey, J. N. Ladd 
and B. E. Volcanl, /. Biol. Chem. 235 480 (I960); H. Weissbach, 
J. N. Ladd, B. E. Volcani, R. D. Smyth and H. A. Barker, ibid. 235 
1462 (I960); J. N. Ladd, H. P. C. Hogenkamp and H. A. Barker, 
Biochem.. and Biophys. Res. Comms. 2 143 (1960). 



Pfizer Handbook of Microbial Metabolites 



446 



groups as shown below. A labeling experiment has shown 
that 



HOOC— CH,— CH,— CO— S— CoA 



CH3 



® 



— C— S— CoA 

migration 

HOOC— CH— CO— S— CoA 

@ — COOH 
,, migration 

HOOC— CH,— CH2— CO— S— CoA 

is the actual process.^" A free radical mechanism was 
proposed in which the Co*""^ of the cobamide coenzyme 
initiates the one-electron transfer: 



COOH 




COOH 


HC— CH3 


Co^±i) ^..^^^ 


^ CHo— CH, 


s— CoA y 


Cobamide 
coenzyme 


/ c=o 

S— CoA 


COOH i 


J 


V COOH 


HC— CH2 y 

1 i/ 


"^ Co<S) + H ® 


• CH— CH2 


c=o 




C=0 



S— CoA 



S— CoA 



This is analogous to a rearrangement reported earlier by 
Urry and Kharasch.-^'^ 

The same organism (Clostridiian tetanomorphiim) was 
found capable of producing coenzymes containing the 
benzimidazole and dimethylbenzimidazole forms of vita- 
min Bjo.'^ The dimethylbenzimidazole coenzyme has been 
found'^' to promote the equilibrium reaiTangement previ- 
ously known to exist:''* 

5° H. Eggerer, P. Overath and F. Lynen, /. Am. Chem. Soc. 82 2643 
(1960). 

SI W. H. Urry and M. S. Kharasch, ibid. 66 1438 (1944). 

^^ H. Weissbach, J. Toohey and H. A. Barker, Proc. Nat. Acad. Sci. 
45 521 (1959). 

''^ E. R. Stadtman, P. Overath, H. Eggerer and F. Lynen, Biochem. 
and Biophys. Res. Comms. 2 1 (I960); Joseph R. Stern and Daniel L. 
Friedman, ibid. 2 82 (I960); Shantov Gurnani, S. P. Mistry and 
B. Connor Johnston, Biochim. et Biophys. Acta 38 187 (1960). 

•'* Robert W. Swlck and Harland G. Wood, Proc. Nat. Acad. Sci. 
U. S. 46 28 (1960). 



447 Pyrroles, Porphyrins and Related Compounds 

CH3 

I 
HOOC— CHo— CH.— CO— CoA ^ HOOC— CH— CO— CoA 

Succinyl Coenzyme A Methylmalonyl Coenzyme A 

The final step in the conversion of succinate to propionate 
is the biotin-dependent decarboxylation : -'^ 




CH3 
Methylmalonyl Coenzyme A Propionyl Coenzyme A 

The total process can be written: 

(1) Acetyl CoA + Succinate ^ Succinyl CoA + Acetate 

B12 coenzyme 

(2) Succinyl CoA , Methylmalonyl CoA 

(3) Methylmalonyl CoA + Biotinenzyme :^ CO2 Biotinenzyme + Propionyl CoA 

(4) Propionyl CoA + Acetate ^ Acetyl CoA + Propionate 

Perhaps it is significant that propionibacteria are rela- 
tively rich sources of vitamin Bjo and of bio tin. This 
scheme also shows how propionic acid can be oxidized by 
entry into the carboxylic acid cycle. 

The precise mechanism by which these interesting rear- 
rangements are promoted by the Bj^ coenzymes remains 
to be determined. It has been pointed out^^ that, in effect, 
what is accomplished is a transpropionation. 

A monograph on vitamin Byj has been published. ^^ 

The cytochromes are heme proteins important in elec- 
tron transport. The most studied is cytochrome c. The 
commonest source is muscle, but yeast cytochrome c has 
been crystallized.'"' Classification is made by spectrum, 
and the proteins are species specific. 

The prosthetic group of cytochrome c is protoporphyrin 
IX bound firmly to the apoenzyme by covalent bonds be- 
tween the thiol groups of cysteine and the vinyl groups of 
the porphyrin.^' Four of the iron coordination bonds are 

^^ E. Lester Smith, "Vitamin Bi-," John Wiley & Sons, Inc., New 
York, 1960. 

°^ Bunji Hagihara, Takekazu Horlo, Kazuo Okunuki, Jinpei 
Yamashita and Mitsuhiro Nozaki, Nature 178 629 (1956). 

^" K. Zeile and H. Meyer, Hoppe-Seylers Z. physiol. Chem. 262 178 
(1939); H. Theorell, Enzymologia 6 88 (1939); Karl-Gustav Paul, 
Acta Chem. Scand. 5 389 (1951). 



Pfizer Handbook o£ Microbial Metabolites 



448 



to porphyrin nitrogen, the other two to histidyl residues 
in the protein. 

Proteolytic enzyme degradation of cytochrome c has 
yielded the polypeptide fragment in the vicinity of the 
porphyrin, and the amino acid sequence has been de- 
termined. It is thought to be:'"'* 



histidyl vaiyl-glutamyl-lysyl-cysteinyl 




alanyl 



glutamyl 



iysyl-glutamyl-vaiyl-threonyi-histidyi-cysteinyl 

Cytochrome c Fragment (Hemopeptide) 

Bovine cytochrome c has a particle weight of about 
13,000 and contains about 20 lysine and 3 or 4 histidine 
residues. A helical model of the Pauling type thus prob- 
ably showns the entire active region of the enzyme since 
this cytochrome contains only one prosthetic group. 

Cytochromes (c^ and Cr,) isolated from Azotobacter 
vinelandii have a particle weight of about 12,000 and con- 
tain 0.46% iron, so that superficially they resemble mam- 
malian cytochrome c.^^ In a comparative study of mam- 
mahan and bacterial (Pseudomonas aerugmosa) cyto- 

s«Hans Tuppy and G. Bodo, Monatshefte Chem. 85 1024, 1182 
(1954); Hans Tuppy and Sven Paleus, Acta Chem. Scand. 9 353, 365 
(1955). 

59 A. Tissieres, Biochem. J. 64 582 (1956). 



449 



Pyrroles, Porphyrins and Related Compounds 



chrome c rather minor spectral differences were noted, 
but there were gross differences in the amino acid com- 
position of the protein.'"'" The prosthetic group of cyto- 
chrome ao from Aerobacter aerogenes has been purified 
but not crystallized/'^ Strict anaerobes such as Clostridia 
seem to lack cytochromes, and some lactobacilli seem to 
use flavins instead. 

Reviews of the role of cytochromes in electron transport 
have been published. ''-• '^- '^*' °^' This process is shown in 
outline in the accompanying diagram. 



TPN + 

TPN— DPN 

Transhy- 

drogenase 



TPNH 



DPNH. 




Succinic 

Dehydrogenase- 

FADH. 



ELECTRON TRANSPORT 

Amytal 
Sensitive 

i 
E-FAD 

T 

DPNH 



Dehy' 
drogenase 

i 
E-FADH-; 




Antimycin Cyanide 

Sensitive Pr Sensitive 

i I i 

E-2-Heme-Fe++ 2-Heme-Fe+++ E-2(Cu+)Heme 
T \ / \/ Fe++ 

Cytochrome y Cytochrome V Cytochrome 



i 



03 
(Oxidase) 




E-2Heme-Fe + + + 2-Heme-Fe++ E-2(Cu++)Heme- H2O 

T I T Fe+++ 

CP T Pr CP 



E-Cytochrome b 

or 
E-Coenzyme Q 



E = Apoenzyme 
CP = Coupled phosphory- 
lation (ATP Synthesis) 
Pr = Protein 



The role of Hpides and quinones in electron transport 
has been discussed. '''' The mechanism of coupled phos- 
phorylation is not understood in detail, but can be repre- 
sented as follows: 



^ Martin D. Kamen and Yoshiro Takeda, Biochim. et Biophys. Acta 
21 518 (1956). 

«i J. Barrett, Biochem. }. 64 626 (1956). 

•52 Albert L. Lehninger, The Harvey Lectures, 49 176-215 (1955); 
idem.. Scientific American 202 102-118 (1960). 

*^^ Britton Chance and G. R. Williams, Advances in Enzymol. 17 
65-130 (1956). 

*'^ Joseph S. Fruton and Sofia Simmons, "General Biochemistry," 
John Wiley and Sons, New York, 1958, pp. 284-386. 

^^ David E. Green and Johan Jarnefelt, Perspectives in Biol, and 
Med. 2 163-184 (1959). 

««D. E. Green and R. L. Lester, Federation Proc. 18 987-1000 
(1959). 



Pfizer Handbook of Microbial Metabolites 450 



H3P04 

Inorganic Phosphate 


XH2 
/a reduced electron\ 
\ carrier / 


X 

/ an oxidized \ 
\electron carrier/ 


ADP H 


ATP 



Some electron transport poisons are shown. Many 
other poisons also act by interfering somehow with the 
function of the electron transport enzymes. 

A lucid, if rather popularized, exposition has been pub- 
lished of the energy relationships in cell respiration, as 
well as the gross cell structure involved.'*^ 

913 Holomycin (Des-N-methylthiolutin), CyH^OoNoSo, orange-yellow 

leaflets, m.p. 264-271° (dec). 

O 

NH— C— CH3 

/ 

s — c=c 

s I I 

CH=C C 

H O 

Streptomyces griseus (Krainsky) Waksman et Henrici 
L. Ettlinger, E. Gaunaann, R. Hiitter, W. Keller-Schierlein, 

F. Kradolfer, L. Neipp, V. Prelog and H. Zahner, Helv. Chim. 

Acta 42 563 (1959). 

914 Thiolutin (Acetopyrrothine, Farcinicin), C8H8O0N0S2, yellow 

crystals, m.p. 260-270° (dec). 









II 






NH- 


II 

-c- 


-CH: 




/ 






s- 


-c — c 






/ 








s 








\ 








c= 


=c c 






H 


1 
CHj 







«7 Albert L. Lehninger, Scientific American 202 102-117 (1960). 



451 Pyrroles, Porphyrins and Related Compounds 

Streptomyces albus 

Walter D. Celmer, Fred W. Tanner, Jr., M. Harfenist, T. M. 
Lees and I. A. Solomons, /. Am. Chem. Soc. 74 6304 (1952). 

Walter D. Celmer and I. A. Solomons, ibid. 77 2861 (1955). 
(Structure) 

915 Noformicin, C,sHi,-ON-, Dihydrochloride m.p. 265° (dec), [a]," 

+ 7.0° (c 1.0 in water). 

[ ] O NH 

HN H C— NH— CHo— CH.2— C— NH2 

Nocardia formica 

Reed A. Gray, Phytopathologij 45 281 (1955). 
Robert L. Peck, Henry M. Shafer and Frank J. Wolf, U. S. 
Patent 2,804,463 (1957). 

916 Aureothricin ( Propiopyrrothine ) , CgHioO^N^So, yellow crystals, 

m.p. 256° (dec). 



NH— C— CH,— CH3 

/ 

s — c=c 

s I I 
\ I I 

CH=C C 

I o 

CH, 

Streptomyces cellulofiavus n. sp. 

Haruo Nishimura, Toshlaki Kimura and Masa Kuroya, /. 
Antibiotics (Japan) 6A 57 (1953). 

Walter D. Celmer and I. A. Solomons, J. Am. Chem. Soc. 77 
2861 (1955). (Structure) 

917 Pyoluteorin, CuH^OgNCL., m.p. 174° (dec). 
Partial structure: 



2 CI 




Pfizer Handbook of Microbial Metabolites 452 

Pseudomonas aeruginosa 

Rokuro Takeda, J. Am. Chem. Soc. 80 4749 (1958). (Struc- 
ture) 

918 Netropsin (Congocidine, Sinanomycin, T1384), CigHoeOgNio, 
the hydrochloride crystallizes as colorless, hygroscopic 
prisms, m.p. 168-172° (dec). 

NH NH 

II II 

H2N— C— CH2— C— NH 



u 



o 

\ll 

I C— NH 
CH3 



O 



o o 

\ll II 

I C— NH— CH2— CH2— C— NH2 
CH3 

Streptomyces netropsis, S. chromogenes n.sp., S. ambo- 
faciens n. sp. 

A. C. Finlay, F. A. Hochstein, B. A. Sobin and F. X. Murphy, 
J. Am. Chem. Soc. 73 341 (1951). 

E. E. van Tamelen and A. D. G. Powell, Chem.. and Ind., 365 
(1957). (Structure) 

919 Prodigiosin, C20H25ON3, red crystals with a green reflex, m.p. 
151.5-152.9° (dec). 
Alternative structures : * 



(CH2)4— CH3 





(CH2)4— CH3 



* See addendum. 



453 Pyiroles, Porphyrins and Related Compounds 

Serratia marcescens (Bacillus prodigiosum), S. marino- 
rubriivi 

Fritz Wrede and Alexander Rothhaas, Z. physiol. Chem. 226 
95 (1934). 

Other metabohtes which have been isolated from cul- 
tures of Serratia marcescens are: 

920 A "prodigiosin precursor," C10H10O0N2, colorless nee- 
dles, m.p. > 250° (dec). 

921 A colorless, crystalline compound, not an antibiotic, 
CS4HG0O10N3, m.p. 153°. 

922 An amide, C04H33O2N7. 
Palmitic acid. 

Three other red, one orange and one blue pigments. 

A polypeptide, marcescin. 

A polysaccharide. 

Fritz Wrede and Alexander Rothhaas, Z. physiol. Chem. 226 
95 (1934). 

Ursula V. Santer and Henry J. Vogel, Biochim. et Biophys. 
Acta 19 578 (1956). 

O. M. Efimenko, G. A. Kuznetsova and P. A. Yakimov, 
Biokhimiya 21 416 (1956). 

A. J. Castro, J. F. Deck, M. T. Hugo, L. R. wmiams and 
M. R. Zingg, /. Org. Chem. 23 1232 (1958). 

A. J. Castro, A. H. Corwin, F. J. Waxham and A. L. Beilby, 
ibid. 24 455 (1959). 

Doris P. Courington and T. W. Goodwin, J. Bacteriol. 70 
568 (1955). 

Harry H. Wasserman, James E. McKeon, Lewis Smith and 
Peter Forgione, /. Am. Chem. Soc. 82 506 (1960). (Structure 
shown above) 

A. Treibs and R. Galler, Angew. Chem. 70 57 (1958). 

923 Celesticetin, C24H3g09N2S, hygroscopic glass, m.p. (Oxalate): 

147-152°, [aU-' +126.6° (c 0.5 in chloroform), [aW* 
(Oxalate) 106.6° (c 0.5 in water). 
Proposed Structure: 

O 

CH2— CH2— O— C— '^^^ 

S / 

HO I HO 

O 



0\ CH-NH-C-L^ 

I CH, '.. I 



"o in ^"^ CH 



CH3 

OCHj 



Pfizer Handbook of Microbial Metabolites 



454 



Streptoviyces celestis n. sp., resembling S. glaucus 

C. DeBoer, A. Dietz, J. R. Wilkins, C. N. Lewis and G. M. 
Savage, "Antibiotics Annual 1954-1955," Medical Encyclope- 
dia, Inc., New York, p. 831. 

Herman Hoeksema, Glen F. Crum and William H. DeVries, 
ibid. p. 837. 

Clarence DeBoer, Alma Dietz and Herman Hoeksema, U. S. 
Patent 2,928,844 (1960). (Structure) 

924 Prodigiosin-Iike Pigment, C^.-.Ha-.ON.j, orange crystals, partial 

melting 147-149°, resolidification, melting 203°. 

Streptomycete related to S. ruber (Krainsky, Waksman 
and Henrici) and S. roseodiastaticus, Waksman and 
Lechevalier 

F. Arcamone, A. DiMarco, M. Ghione and T. Scotti, Giorn. 
microbiol. 4 77 (1957). 

925 Hematin, C34H3oO4N4Fe®OH0. 



CH=CH 




CH=CH2 



HOOC— CH2— CH 



CH2— CH2— COOH 



Saccharomyces anamensis 

H. Fischer and F. Schwerdtel, Z. physiol. Chem. 175 248 
(1928). 

926 Protoporphyrin, C34H34O4N4, deep red crystals, m.p. >300°. 

CH^CH, CH3 



CH=^CH2 




HOOC— CH2— CH2 



CHo— CH2— COOH 



455 Pyrroles, Porphyrins and Related Compounds 

Yeasts, Rliodopseudomonas spheroides, other photosyn- 
thetic bacteria 

Hans Fischer and Hermann Fink, Z. physiol. Chem. 140 57 
(1924). 

927 Coproporphyrin I, Cno,H3s08N4. 



HOOC— CH,— CH 



CHo— CH2— COOH 
CH3 




CH2— CH2— COOH 



CH2— CH2— COOH 



Saccharomyces cerevisiae, S. anamensis, other yeasts, 
Aspergillus oryzae, photosynthetic bacteria 

Hans Fischer and Hermann Fink, Z. physiol. Chem. 150 243 
(1925). 

928 Coproporphyrin III, C36H3SO8N4, dark red crystals. 



CH2— CHo— COOH 
CH3 




CHo— CH,— COOH 



HOOC— CHo— CHo 



CH2— CHo— COOH 



Mycobacterium tuberculosis var. hominis, Rhodopseu- 
domonas spheroides, Corynebacterium diphtheriae 

M. O'L. Crowe and A. Walker, Brit. J. Exptl. Path. 32 1 
(1951). 

C. M. Todd, Biochem. J. 45 386 (1949). 



Pfizer Handbook of Microbial Metabolites 456 

929 Uroporphyrin III, C40H38O16N4. 

:— OH ^ 

CH2— C— OH 



CH2— CHo— C— OH / 




CH2— CH2— C— OH 



CH2— C— OH 

/ 

CH2 CHo— CHo— C— OH 

Rhodopseudomonas spheroides 

June Lascelles, Abstracts Gordon Research Conference, 
Vitamins and Metabolism (1958). (Detection) 

H. Fischer and H.-J. Hofmann, Z. physiol. Chem. 246 15 
(1937); H. Fischer and A. Miiller, ibid. 246 31 (1937). (Struc- 
ture) 

930 Bacteriochlorophyll a, C55H740(;N4Mg, amorphous, slow decom- 
position above 94°. 



0=C— CH3 cHs H 




C20H39— OOC CH2— CH 
Phytyl H3COOC 

Rhodospirillum rubrum, R. fulvum, Rhodopseudomonas 
spheroides, Thiocystis violacea, other Rhodovibrio spp. and 
sulfur and chlorobacteria 

Hans Fischer and Robert Lambrecht, Z. physiol. Chem. 249 
1 (1937). 

Hans Fischer, Robert Lambrecht and Hellmuth Mittenzwei, 
ibid. 253 1 (1938). 

John W. Weigl, /. Am. Chem. Soc. 75 999 (1953). 

A. Seybold and G. Hirsch, Naturwissenschaften 41 258 
(1954.) 



457 



Pyrroles, Porphyiins and Related Compounds 



931 Vitamin Bi^ (Cyanocobalamin, a-(5,6-Dimethylbenzimidaz0lyl) 
cobamide cyanide), Ce3HssOi4Ni4PCo, dark red crystals 
which blacken near 212° and do not melt below 320°, 
[a]6563"^ —59 ±9° (dilute aqueous solution). 



NH2COCH2 CH.CHoCONHz 




CH2CONH2 
CH2CH2CONH2 



HOCH2 



Vitamin B12 activity has been detected in fermentation 
broths from many microorganisms, e.g. Streptomyces 
griseus, S. antibioticus, S. roseochromogenes, Mycobacte- 
rium smegmatis, Lactobacillus arabinosus, propionibacte- 
ria. Crystalline material has been isolated from some of 
these. For primary fermentations, Streptomyces olivaceus 
is probably the best producer (3.3 mg. per liter). 

Dorothy Crowfoot Hodgkin, Jennifer Kamper, Maureen 
MacKay and Jenny Pickworth, Nature 178 64 (1956). (Struc- 
ture) 

W. H. Sebrell, Jr. and Robert S. Harris, "The Vitamins," 
Robert S. Harris, Donald E. Wolf, Karl E. Folkers, H. M. 
Wuest, Thomas H. Jukes and William L. Williams, Vitamin B12, 
Academic Press Inc., New York, 1954 Vol. I Chap. 3, pp. 396- 
524. (A review) 



Pfizer Handbook of Microbial Metabolites 458 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
1954 Vol. II, J. M. VanLanen, Production of vitamins other 
than riboflavin, chap. 6, pp. 207-8. 

932 Factor B is vitamin Bjo from which the nucleotide 

moiety has been removed. It has been isolated from 
fermentations, from rumen contents, from sev^^age, and 
it can be prepared chemically from vitamin B^o. 

E. Lester Smith, "Vitamin B12," John Wiley and Sons, Inc., 
New York, 1960, 196 pp. (A monograph) 

This monograph also explains the new nomenclature 
system for B^o and related compounds. 

Other intermediates in the biosynthesis of vitamin B12 
by Propionibacterium shermanii have been detected: 

Konrad Bernhauer, Elisabeth Becher, Gisela Gross and 
Georg Wilharm, Biochem. Z. 332 562 (1960). 

K. Bernhauer, Hw. Dellweg, W. Friedrich, G. Gross, F. 
Wagner and P. Zeller, Helv. Chim. Acta 43 693 (1960). 

K. Bernhauer, F. Wagner and P. Zeller, ibid. 43 696 (I960). 

g. INDOLES 

The indole nucleus occurs in microorganisms in such 
forms as tryptophan, one of the less abundant amino 
acids, in bacterial pigments such as violacein and indigo 
and in amines from higher fungi such as serotonin and 
psilocybin, which have strong physiological effects in 
higher animals. The indole nucleus is incorporated also 
into bizarre fungal metaboUtes such as echinulin and 
gliotoxin, into the mushroom poisons, such as phalloidin, 
and into the ergot alkaloids listed in the following section. 

One route to indole and to tryptophan was outlined in 
the section on amino acids. This is the pathway discov- 
ered by Yanofsky and confirmed and elaborated in his and 
other laboratories.^ Anthranilic acid from the shikimic 
acid route combines with ribose phosphate, cyclization oc- 
curs to form the pyrrole ring, a triose phosphate is elimi- 

^ C. Yanofsky, Biochim. et Biophys. Acta 16 594 (1955); idem., 
J. Biol. Chem. 223 171 (1956); F. Gibson, M. Jones and H. Taltscher, 
Biochem. J. 64 132 (1956); P. A. Trudinger, ibid. 62 480 (1956); 
F. Lingens and H. Hellmann, ATigew. Chem. 69 97 (1957); L. W. 
Parks and H. C. Douglas, Biochim. et Biophijs. Acta 23 207 (1957); 
J. Gots and S. Ross, ibid., 24 429 (1957); C. Yanofsky and M. Rach- 
meier, ibid. 28 640 (1958). 



459 



Indoles 



nated and the indole so formed combines with L-serine to 
fonn L-tryptophan : 



COOH 



COOH 




r^ 




NH' 



CH— CH— CH— CH— CH.— O— PO3H2 

' I 1 

NH. OH OH 

Anthranilic Acid N-(2-Carboxyphenyl)-l -aminoribose-5-phosphate 

CH— CH- CH, 



J 



' \ I I Triose phosphate 
OH OH O— POaHj ^ 



'OO 



L-Serine 



lndolyl-3-glycerol Phosphate 



a? 



CHo— CH— COOH 
NHo 



L-Tryptophan 

N-Fructosylanthranilic acid has been isolated from a 
yeast, and it may be another intennediate in indole syn- 
thesis. In this case a tetrose would be eliminated. If 
pentoses and hexoses can both be used in reactions with 
anthranilic acid, perhaps tetroses can be as well. This 
possibility is emphasized by Wenkert" in a discussion of 
alkaloid biosynthesis. A reaction of this sort might ex- 
plain the frequent occurrence in nature of indole deriva- 
tives with two carbon atom side-chains in the 3-position. 
In other words the indole biosynthesis could be general- 
ized: 




-Ernest Wenkert, Experientia 15 165 (1959). 



Pfizer Handbook of Microbial Metabolites 460 




H 

in which R — CH — CH — CHO is any of several sugars. 

OH OH 

It may be that other derivatives of anthranihc acid can 
participate in this route, too. For example 5-hydroxyan- 
thranilic acid would give rise to the 5-oxyindole derivatives 
found in nature. It is notable that this acid is a growth 
promoter for an Escherichia coli mutant.^ 

Ascorbigen, a bound form of ascorbic acid isolated from 
plants of the cabbage family, has one of the structures:* 

CH2— C^=CH CH2— C=CH 



oa< > CD 



H 



HO— C CH or H CH C— OH 

/ \ 

CHo— CH— CH C=0 0==C CH— CH— CH2 



OH OH \^/ \^/ OH OH 



O 



Ascorbigen 
(alternate structures) 



The presumptive precursor is 3-indolylacetol, analogous to 
an intermediate in histidine biosynthesis, and it is interest- 
ing to speculate as to whether this is an offshoot of the 
biosynthetic route to tryptophan or whether it is formed by 
way of tryptophan. 

The mold product, echinuhn, has an unusual structure, 

apparently involving the indole synthesis, terpenoid and 

amino acid precursors. Gliotoxin, on the other hand, is 

almost entirely derived from amino acids, and it could 

have been classified as a polypeptide. C"-Labeling studies 

have demonstrated the following biosynthetic pathway for 

gliotoxin : ^ 

3H. Niemer and A. Oberdorfer, Z. physiol. Chem. 308 51 (1957). 

■* Z. Prochazka, V. Sanda and F. Sorm, Coll. Czech. Chem. Comm. 

22 654 (1957). 

s J. A. Winstead and R. J. Suhadolnik, J. Am. Chem. Soc. 82 1644 
(1960); R. J. Suhadobiik, A. Fischer and J. Wilson, Federation Proc. 
19 8 (1960). 



461 



Indoles 



H,N^\ 

COOH 

Phenylalanine 



lO] 



H-iU^^ 



Serine 



COOH Methionine 
m-Tyroslne 




XH3 
CH2OH 

''from Serine^ 

and 
^Methionine 
Dethiogiiotoxin 



2S 



N s 
s N 



,/- 



CH2OH 
Gliotoxin 



Methionine was the most efficient source of the N-methyl 
group, the ^-carbon of serine being about one third as ef- 
fective. Both of the amino acid skeletons were incorpo- 
j rated intact when furnished, and m-tyrosine could also be 

used as a precursor. 

, 933 Indole, CgHyN, colorless leaflets, m.p. 52°. 



cxj 



934 



H 

Escherichia coli mutants, yeasts, Treponema spp. 

P. A. Trudinger, Biochem. J. 62 480 (1956). 

Charles Yanofsky, /. Biol. Chem. 223 171 (1956). 

F. Gibson, Marjorie J. Jones and H. Teltscher, Biochem. J. 
64 132 (1957). 

L. W. Parks and H. C. Douglas, Biochim. et Biophys. Acta 
23 207 (1957). 

Michel Moureau and W. Aladame, Ann. inst. Pasteur 88 231 
(1955). 

Indole-3-acetic Acid (Rhizopin), CioHgOoN, colorless plates, m.p. 
164°. 



oa 



CHoCOOH 



Rhizopus suinus, R. nigricans, Aspergillus niger, Peni- 



Pfizer Handbook of Microbial Metabolites 462 

cillium notatum, Absidia ramosa, Boletus edulls, Yeasts 

Niels Nielsen, Biochem. Z. 237 244 (1931); 249 196 (1932). 

Fritz Kogl and D. G. F. R. Kostermans with A. J. Haagen- 
Smit and H. Erxleben, Z. phijsiol. Chem. 228 113 (1934). 

Kenneth V. Thimann, /. Biol. Chem. 109 279 (1935). 

Donald J. Cram and Max Tishler, /. Am. Chem. Soc. 70 
4238 (1948). (Isolation) 

Ryuichi Honda, Japanese Patent 603 (1950). 

935 Serotonin ( 5-Hydroxytryptamine ) , C10H12ON2 (Hydrochloride), 
colorless crystals, m.p. 167°. 

HO CH2CH2NH2 



Oj 



H 

Panaeolus campanulatus 

Demonstrated by paper chromatography only. 

Varro E. Taylor, Jr., Science 128 718 (1958). 

936 Psilocin, CjsHigONs, colorless crystals, m.p. 173-176° (dec). 



H 
O 




CH2CHoN(CH3 



Psilocybe species 

Psilocin is a minor constituent of the mushrooms which 
contain psilocybin. 

A. Hofmann and F. Troxler, Experientia 15 101 (1959). 

937 Psilocybin, C12H17O4N2P, colorless crystals, m.p. 185-195° 
(dec). 

OH 

o=p— o® 
o 



OJ 



CH3 

@/ 

CH2CH2N 

\ 

CH3 



463 Indoles 

Psilocybe mexicana Heim, P. caerulescens Murr. var. 
Mazecotorum Heim, P. aztecorum Heim, P. sempervirens 
Heim et Cailleux, P. zapotecorinn Heim, Stropharia cuhen- 
sis Earle 

A. Hofmann, R. Heim, A. Brack and H. Kobel, Experientia 
14 107 (1958). 

A. Hofman, A. Frey, H. Ott, Th. Petrzilka and F. Troxler, 
ibid. 14 397 (1958). (Synthesis) 

938 Gliotoxin ( Aspergillin ) , Ci:^Hi403NoSo, m.p. 195° (dec), [aW-' 
-290° ±10° (c 0.078 in ethanol). 



/ 



s 



O 



CH3 
CH2OH 



Trichoderma viride, Aspergillus fumigatus, Penicillium 
terlikowski Zaleski, P. cinerascens, P. jenseni, Gliocladium 
fimbriatuTn 

The yield of gliotoxin and its acetate from P. terlikowski 
Zaleski was reported as about 100 mg. per liter. 

John R. Johnson, William F. Bruce and James D. Dutcher, 
J. Am. Chem. Soc. 65 2005 (1943) and other papers in this 
series. 

Malcolm R. Bell, John R. Johnson, Bernard S. Wildi and 
R. B. Woodward, J. Am. Chem. Soc. 80 1001 (1958). (Struc- 
ture) 

939 Gliotoxin Acetate, Ci-.HieO-NoS^, pale yellow rhombic crystals, 
m.p. 159°, [a]i,''' -197° (c 0.600 in chloroform). 




^N 



O 

O^^CH3 

CH2— O— C— CH3 



Penicillium terlikowski Zaleski 

John R. Johnson, Aklaq R. Kidwai and John S. Warner, /. 
Am. Chem. Soc. 75 2110 (1953). 



Pfizer Handbook of Microbial Metabolites 464 

940 Indigo, C16H10O2N2, blue powder with a coppery luster, sublimes. 



Schizophyllum commune mutant 
Ammonium ion was the only nitrogen source. 
Philip G. Miles, Henning Lund and John R. Raper, Arch. 
Bioche7n. and Biophys. 62 1 (1956). 

941 Chetomin, Ci(5Hi704N3S2 (proposed), amorphous white powder, 

m.p. 218-220° (dec), [ajn'^ +360° (c 1 in chloroform). 

A neutral compound. Positive indole, Hopkins-Cole, 
negative biuret, Millon. 

Chaetomium cochlioides 

Walton B. Geiger, Jean E. Conn and Selman A. Waksman, 
;. Bacterial. 48 531 (1944). (Isolation) 

Walton B. Geiger, Arch. Biochem. 21 125 (1949). 

942 Violacein, C20H13O3N3, violet-black microcrystals, m.p. >350° 

(dec). 

HO C=CH 



Co 



NH C 

c ^1 — 

O X N 
O H 



Chromohacterium, violaceum. 

F. M. Strong, Science 100 287 (1944). 

R. T. S. Beer, Angew. Chem. 69 676 (1957). 

J. A. Ballantine, C. B. Barrett, R. J. S. Beer, B. G. Boggiano, 
K. Clarke, Stephen Eardley, B. E. Jennings and Alexander 
Robertson, /. Chem. Soc, 2222 (1957) and preceding papers 
in this series. 

J. A. Ballantine, R. T. S. Beer, D. J. Crutchley, G. M. Dodd 
and D. R. Palmer, /. Chem. Soc, 2292 (1960). (Synthesis) 

R. D. Demoss and N. R. Evans, /. Bacteriol. 79 729 (1960). 
(Biosynthesis) 



465 Ergot Alkaloids 

943 Echinulin, CosHa^OsNg, white needles, m.p. 242°. 
Probable structure: 




Aspergillus glaucus types, A. echinulatus, A. chevalieri 
About 200 g. of pure material were obtained from 5 kg. 

of dry mycelium. Auroglaucin and flavoglaucin were 

isolated from the same source. 

A. Quilico and L. Panizzi, Ber. 76B 348 (1943). (Isolation) 
Adolfo Quilico, Cesare Cardini and Franco Piozzi, Gazz. 

Chim. ital. 86 211 (1956). (Structure) 

Ziro Kitamura, Uzukiko Kurimoto and Matatsugu Yoko- 

yama, 7- Pharm. Soc. Japan 76 972 (1956). 

C. Cardani, G. Casnati, F. Piozzi and A. QuUico, Tetrahedron 

Letters No. 16 1 (1959). (Structure) 

h. ERGOT ALKALOIDS 

The constituents of the sclerotia of the fungus Claviceps 
purpurea (Fries) TuL, a cereal parasite, have been exten- 
sively studied. Some of the alkaloids are used in medicine 
for their oxytocic properties and to relieve migraine. 

Ergocristine, ergokryptine and ergocornine (and their 
isomers) constitute a closely related complex formerly 
thought to be homogeneous and called ergotoxine. Be- 
sides the alkaloids which are shown in the succeeding 



Pfizer Handbook of Microbial Metabolites 



466 



pages, many other chemicals have been identified. 
Among them are: 



Ergothioneine 

Histidine 

Tyrosine 

Betaine 

ChoUne 

Acetylcholine 

Cadaverine 

Putrescine 

Agmatine 

Histamine 

Tyramine 

Valine 



Leucine 

Ammonia 

Methylamine 

Trimethylamine 

Ethylamine 

n-Propylamine 

iso-Propylamine 

iso-Butylamine 

iso-Amylamine 

n-Hexylamine 

ytJ-Phenylethylamine 

Mannitol 



Clavicepsin 
Ergosterol 
Oils 

Lactic Acid 
Succinic Acid 
Oxalic Acid 
Citric Acid 
Formic Acid 
Ethanol 
Furfural 
Acetaldehyde 
Acetone 

Ergoflavine and 
other pigments 



Careful work has shown that many of the alkaloids 
produced in the natural state can be produced in artificial 
culture as well/ -• ■' Total alkaloid yields of 1000-1500 
mg. per liter of culture fluid have been obtained exclusive 
of mycelial alkaloids.^ 

The conventional ergot alkaloids contain the lysergic 
acid moiety I or isolysergic acid, the stereoisomer at posi- 
tion 8. 



HOOC H 




^ A. Hofmann, R. Brunner, H. Kobel and A. Brack, Helv. Chim. 
Acta 40 1358 (1957). 

'W. A. Taber and L. C. Vining, Can. J. Microbiol. 3 55 (1957). 
3 Ervin Glaz, Acta Pharm. Hung. 25 11 (1955). 



I 



467 



Ergot Alkaloids 



A number of different hypotheses have been advanced 
concerning the biosynthetic origin of the ergot alkaloids. 
These are outlined below: 
( 1 ) van Tamelen ( 1 953 ) : * 



HO 



r^ 




; 



CHo— CH— COOH 



NH2 



cXj 



lO] o^ 



CH — CH— COOH 

' ' I 

NH2 + 



I 
CH3 



CONHo 



CHo— CH— COOH 

I 
NH2 




CONH2 

(/ N— CH3 
V CH.- 


C0NH2 

^ N— CH 

-CH— COOH — ^ 

1 H2 J\ / 


kJ^N^ 


WH 1 ' 1 1 


NM2 ' 1 1 
— H2O ^^:^>^N^ 
H 



CH>— CH— COOH 

I 
NHo 



CONHo 




CH2— COOH 



© 



— H2 



< 



CONH2 

I 



N— CH3 



— CO2 ^ 




N 
H 

i 
Lysergic Acid 



(2) Harley-Mason (1954):^ 

Eugene van Tamelen, Experientia 9 457 (1953). 
'J. Harley-Mason, Chem. and Ind., 251 (1954). 



Pfizer Handbook of Microbial Metabolites 



COOH 



CH2O 



CH2 / 



468 



0=C NH2 

\ / 

HOOC— CH2 C— COOH 

\ 
CH2 



^ 



COOH 



NH 







"^^X^ 



N' 



COOH 



)0= 




NH 



Lysergic Acid 



HOC 


(3) Wendler (1954) :« 

NH2 
/ 
^^-^\ NH- 

\h2 >/ 
/ -H2O ^ > 

1" 1 ) ' 1 II 


-R 


COOH 

1 
HO— C— CH2— COOH 

1 
CH2 

/ 


k;^ 


1 L i 1^ 

H H 


HOOC 




(Citric Acid) 



- HO COOH 

\l 
^^^^ /C— CH2— COOH 
HOOC^<: NH— R 

HO- 




COOH 




:CH2 

NH— R 



Lysergic Acid 



6N. L. Wendler, ExpeHentia 10 338 (1954). 



469 Ergot Alkaloids 

(4) Robinson (1955):^ 

COOH 

I 

CH, 
/ COOH 

CH, I 

\ CH. 

COOH / 

NH— CHj CHj NH— CH3 

/ \ / 

CH— COOH C— CH 

\ II \ 

CH2 O CHj 



H H 




lysergic Acid 



(5) Feldstein (1956):« 

NH, 

"°°^-^( O NH, 

""CH, % / 

oi-Keto- 



a/ 



f^y^ ^Si/^N glutaric Acid 

H H 

COOH 

I 
O CH, 

II / 
HOOC— C— C NH, 




COOH 

I 

CH, 
/ 
HOOC— C NH, 

% / CH2O 



^;?C__J> > Lysergic 

f fl -CO, 

^^=5iA-N-^ +CH, 



^ Sir Robert Robinson, "The Structural Relations of Natural Prod- 
ucts," Oxford University Press, Oxford, 1955. 
8 A. Feldstein, Experientia 12 475 (1956). 



Pfizer Handbook of Microbial Metabolites 470 

(6) Birch (1958),^ Mothes, et aZ. (1958):" 
C 



Isoprene 
Equiva- " 
lent 



C=C 



c 

\ 

c 



NHCH31 



"-N 



J 



Tryptamine — > — > — > Lysergic Acid 
Equivalent 



Each of these hypotheses has had its votaries, but now 
experimental work is beginning to accumulate. There 
have been conflicting results, partly because some experi- 
menters have injected labeled precursors into infected rye 
plants, while others added them to cultures grown on arti- 
ficial medium. 

The 5-hydroxytryptophan proposals have been criti- 
cized^ because no 5-hydroxyindole analogues of lysergic 
acid have been found in nature, and because (obviously 
the devices of organic chemists) they suffer from some 
rather improbable biological intermediates. Brady has 
found that in artificial culture tryptophan was an efficient 
precursor for the clavine alkaloids, while 5-hydroxytrypto- 
phan was not.^^ 

By using parasitic cultures one group reported good in- 
corporation of /;j-C^*-tryptophan,^" while another reported^- 
only weak labeling of the alkaloids isolated from the 
sclerotia. 

By use of a cell homogenate technique, it was found 
that alanine and phenylalanine were incorporated into 
ergotamine and the ergotoxine complex, but not into 
ergonovine, which suggests that these amino acids are 
precursors of the peptide structure of the water-insoluble 

^ G. E. Wolstenholme and Cecilia M. O'Connor, CIBA Foundation 
Symposium on "Amino Acids and Peptides with Antimetabolic Ac- 
tivity," A. J. Birch and Herchel Smith, Oxidative formation of bio- 
logically active compounds from peptides, Little, Brown and Co., Bos- 
ton, 1958, pp. 254-256. 

'^' K. Mothes, F. Weygand, D. Groger and H. Grisebach, Z. Natur- 
forsch. i;}b 41 (1958). 

" Lynn Robert Brady, Dissertation Abstr. 20 2526 (1960). 

'^ R. J. Suhadolnik, L. M. Henderson, J. B. Hanson and Y. H. Loo, 
;. Am. Chem. Soc. 80 3153 (1958). 



471 Ergot Alkaloids 

ergot alkaloids.'-* Ci4-Labeled indole and serine, alone or 
together, were not incorporated. 

Another artificial culture study in which the Claviceps 
purpurea culture was grown saprophytically on a simple 
galactose, ammonium succinate, mineral salts, biotin me- 
dium to which D,L-/^-C"-tryptophan was added, found that 
the tryptophan was an efficient precursor.^' Labeling was 
about the same throughout the range of alkaloids isolated, 
thus suggesting a common biogenesis. Supplementation 
with L-tryptophan increased the yield and caused the for- 
mation of elymoclavine and agroclavine, which were not 
formed otherwise. 

Another (non-tracer) experiment in artificial culture 
showed no increase in total alkaloid production on supple- 
mentation with either tryptophan, hydroxytryptophan, in- 
dole, 5-hydroxyindole or serotonin.^'' 

The consensus of the labeling experiments seems to be, 
however, that tryptophan is a rather direct precursor of 
the lysergic acid skeleton. 

Apparently there is no good evidence yet concerning the 
origin of the remainder of the skeleton. The isoprenoid 
precursor hypothesis is under investigation. •'• ^"^ This pro- 
posal is buttressed by the structure of the mold metabolite, 
echinulin, which has an indole nucleus bearing isoprenoid 
attachments. 

CH3 

\ 

C=CH— CH, 
CH3 r 1^ 11 CH3 CH.OH 



'CU... 



H 



^ C— CH=CH. C— CH3 

CH3 CH2 1 /" 

\ I CH3 HC NH-CH3 

c=c ^ 



CH3 .CHv O 



HN 

I 
^C NH 



CH3 

Echinulin 



kJ.J 



> 



H 

Chanoclavine 



" Aro Garo Paul, Dissertation Abstr. 17 2143 (1957). 

i*W. A. Taber and L. C. Vining, Chem. and Ind. 1218 (1959). 

1^ Ross M. Baxter, S. I. Kandel and A. Okany, Nature 185 241 
(1960). 

^^ A. J. Birch, B. J. McLoughlin and Herchel Smith, Tetrahedron 
Letters No. 7 1 (1960). 



Pfizer Handbook of Microbial Metabolites 472 

It is also supported by the structure of chanoclavine, 
which seems to be not too remotely derived from such an 
intermediate. 

A thorough review of the chemistry of the ergot alka- 
loids has been published. ^^ 

944 Agroclavine, CieH^gNo, colorless crystals, m.p. 210-212° (dec), 
[aW -183° (c 1 in pyridine). 



N— CH3 




Claviceps purpurea (Fries) Tul. 

A. Hofmann, R. Brunner, H. Kobel and A. Brack, Helv. 
Chim. Acta 40 1358 (1957). 

945 Setoclavine, CieHjgON^, colorless crystals, m.p. 229-234° (dec), 

[aW° +174° (c 1 in pyridine). 

946 Isosetoclavine (Triseclavine), CieHigONa (stereoisomer of seto- 

clavine), colorless crystals, m.p. 234-237° (dec), [c(]u~° 
-J-107° (c 1 in pyridine). 




Clavicepts purpurea (Fries) Tul. 

A. Hofmann, R. Brunner, H. Kobe! and A. Brack, Helv. 
Chim. Acta 40 1358 (1957). 

" Arthur Stoll, Fortschr. Chem. org. Naturstoffe 9 114-170 (1952). 



473 Ergot Alkaloids 

947 Elymoclavinc, CksHi^ONo, colorless crystals, m.p. 245-247° 
(dec), hW -152° (c 1 in pyridine). 



CH,OH 




Claviceps purpurea (Fries) Tul. 

A. Hofmann, R. Brunner, H. Kobel and A. Brack, Helv. 
Chim. Acta 40 1358 (1957). 

948 Penniclavine, CjoHigOoNs, colorless crystals, m.p. 222-225° 

(dec), [aJD'" +153° (c 1 in pyridine). 

949 Isopenniclavine, CicHi^O^No (stereoisomer of penniclavine), 

colorless crystals, m.p. 163-165° (dec), [aW° +146° (c 1 
in pyridine). 

HOCHo 

:n— CH3 




Claviceps purpurea (Fries) Tul. 

A. Hofmann, R. Brunner, H. Kobel and A. Brack, Helv. 
Chim. Acta 40 1358 (1957). 

950 Dihydroagroclavine (Festuclavine), CigHooNo, colorless crystals, 
m.p. 242° (dec), [a]D-° -69° (c 0.5 in chloroform). 




Pfizer Handbook of Microbial Metabolites 474 

Claviceps purpurea (Fries) Tul. 

Matazo Abe, Ann. Rept. Takeda Res. Lab. 10 73, 83, 90, 
110, 126, 129, 145, 152, 167, 171, 179, 190, 205, 210 (1951). 

Matazo Abe, Togo Yamano, Yoshiharu Kozu and Mitsugu 
Kusumoto, ;. Agr. Chem. Soc. Japan 24 416, 471 (1951); 25 
458 (1952); 27 18, 613, 617 (1953). 

Matazo Abe ibid. 28 44, 501 (1954). 

Matazo Abe, Togo Yamano, Yochiharu Kozu and Mitsugi 
Kusumoto, ibid. 29 364 (1955). 

Matazo Abe, Saburo Yamatodani, Togo Yamano and Mit- 
sugi Kusumoto, Bull. Agr. Chem. Soc. (Japan) 19 92 (1955). 

Saburo Yamatodani and Matazo Abe, ibid. 19 94 (1955). 

951 Pyroclavine, CuiH^oNi., colorless crystals, m.p. 204° (dec), [(x]d-° 

-90° (c 0.2 in pyridine). 

and 

952 Costaclavine, CjeHooNo, colorless crystals, m.p. 182° (dec), 

[a] I,'-" +44° (c 0.2 in pyridine). 

These are thought to be isomers of dihydroagroclavine. 

Claviceps purpurea (Fries) Tul. 

Matazo Abe, Saburo Yamatodani, Togo Yamano and Mit- 
sugi Kusumoto, Bull. Agr. Chem. Soc. (Japan) 20 59 (1956). 

953 Dihydroelymoclavine, Ci.jH.oONo, colorless crystals, m.p. 210° 

(dec), [a]^'^ -167° (c 0.16 in chloroform). 



H CH2OH 




Claviceps purpurea (Fries) Tul. 

See references under dihydroagroclavine. 



475 Ergot Alkaloids 

954 Chanoclavine (Secaclavine), Ci,jH^.„ON^., colorless crystals, m.p. 
220-222° (dec), [a],r" -240° (c 1 in pyridine). 

CH2OH 

I 

C— CH3 

/ 
HC NHCH3 




Claviceps purpurea (Fries) Tul. 

A. Hofmann, R. Brunner, H. Kobel and A. Brack, Helv. 
Chim. Acta 40 1358 (1957). 

Matazo Abe, Togo Yamano, Saburo Yamatodani, Yoshiharu 
Kozu, Mitsugi Kusumoto, Hajime Koinatsu and Saburo Ya- 
mada. Bull. Agr. Chem. Soc. (Japan) 23 246 (1959). 

955 Ergobasine (Ergometrine, Ergonovine, Ergotocine, Ergostetrine, 

Ergotrate, Ergoclinine ) , Ci.,Hj;^02N.{, colorless crystals, 
m.p. 162°, [a]ir" +90° (c 1 in water). 

956 Ergobasinine, Ci9H2.s02N;{ (stereoisomer of ergobasine), colorless 

crystals, m.p. 196°, [ajo'" +414° (c 1 in chloroform). 

CH3 

O NH— CH— CHoOH 

\ 



C 




N— CH3 



Claviceps purpurea (Fries) Tul. 

Walter A. Jacobs and Lyman C. Craig, Science 82 16 
(1935). (Structure) 



Pfizer Handbook of Microbial Metabolites 



476 



957 Ergosecalinine, C04H28O4N4, colorless crystals, m.p. 217° (dec), 
[(xW^ +298° (c 0.2 in chloroform). 




o=c 




NHo^ H 



N— CH3 



Claviceps purpurea 

Matazo Abe, Togo Yamano, Saburo Yamatodani, Yoshiharu 
Kozu, Mitsugi Kusumoto, Hojime Komatsu and Saburo Ya- 
mada, Bull. Agr. Chem. Soc. (Japan) 23 246 (1959). 

958 Ergosine, C30H37O5N5, colorless crystals, m.p. 228° (dec), [<x]d^^ 

— 179° (c 1 in chloroform). 

959 Ergosinine, C30H37O5N5 (stereoisomer of ergosine), colorless 

crystals, m.p. 228° (dec), [a]D-° +420° (c 1 in chloro- 
form ) . 




Claviceps purpurea (Fries) Tul. 



477 



Ergot Alkaloids 



A. Stoll, A. Hofmann and Th. Petzilka, Helv. Chim. Acta 34 
1544 (1951). (Structure) 

960 Ergocornine, C3iH390-,N.-,, colorless crystals, m.p. 182-184° 

(dec), [a]D'" -188° (c 1 in chloroform). 

961 Ergocorninine, C31H39O5N5 (stereoisomer of ergocornine), color- 

less crystals, m.p. 228° (dec), [a]u'" +409° (c 1 in chloro- 
form). 




Claviceps purpurea (Fries) Tul. 

A. Stoll, A. Hofmann and Th. Petzilka, Helv. Chim. Acta 
34 1544 (1951). (Structure) 

962 Ergokryptine, C30H41O5N5, colorless crystals, m.p. 212-214° 

(dec), [alD'" -187° (c 1 in chloroform). 

963 Ergokryptinine, CgoH^iOjNg (stereoisomer of ergokryptine) col- 

orless crystals, m.p. 240-242° (dec), [aW +408° (c 1 in 
chloroform ) . 




Pfizer Handbook of Microbial Metabolites 



478 



Claviceps purpurea (Fries) Tul. 

A. Stoll, A. Hofmann and Th. Petzilka, Helv. Chim. Acta 
34 1544 (1951). (Structure) 

964 Ergotamine, C^gH^-.Or.Nr,, colorless prisms, m.p. 212-214° (dec.)> 

[<xW" -160° (c 1 in chloroform). 

965 Ergotaminine, C;^;{H3-,0-,N-, (stereoisomer of ergotamine), color- 

less plates, m.p. 241-243° (dec), [a],/-" +369° (c 0.5 in 
chloroform ) . 




Claviceps purpurea (Fries) Tul. 

Walter A. Jacobs and Lyman C. Craig, /. Org. Chem. 1 245 
(1936). 

Arthur Stoll, Helv. Chim. Acta 28 1283 (1945). 

966 Ergocristine, Cg-.HjiyOr.N-,, colorless crystals, m.p. 165-170° 

(dec), [a]n-" -183° (c 1 in chloroform). 

967 Ergocristinine, Cjj-.HagOr.N.- (stereoisomer of ergocristine), m.p. 

226° (dec), [a],.'" +336° (c 1 in chloroform). 



X^o^jAn-^ 




479 Pyridines 

Claviceps purpurea (Fries) Tul. 

A. Stoll, A. Hofmann and Th. Petzilka, Helv. Chim. Acta 
34 1544 (1951). (Structure) 

i. PYRIDINES 

Few pyridines are listed, but two of these, nicotinic acid 
and pyridoxine, are vitamins. Fusaric acid is a wilt toxin, 
and 2,6-dipicolinic acid appears in conspicuous quantities 
in bacterial spores. 

Dipicolinic acid' - ' ' probably is formed by cyclization 
of ci;,€-diaminopimelic acid, a lysine precursor and cell wall 
constituent of some bacteria: 




HOOC — k J — COOH /^^,, 
NH> NH.. / N' 

HOOC COOH 

Diaminopimelic Acid 2,6-Dipicolinic Acid 

The metaboHc significance, if any, is unknown. In Bacil- 
lus sphaericus diaminopimelic acid is present in spores 
and not in vegetative cells, but in many bacteria it is pres- 
ent in both. 

Fusaric and dehydrofusaric acids are by-products of the 
gibberellin fermentation and are produced by fusarium 
types. These include plant pathogens, and fusaric acid 
solutions sprayed on healthy plants of the usual host 
cause wilting typical of infection. Apparently no study 
has been made of the mode of biogenesis. 

Nicotinic acid in its coenzyme forms occurs in all living 
cells where it is essential in hydrogen and electron trans- 
port. It is used by a variety of apoenzymes as the pros- 
thetic group for various dehydrogenase reactions. It is 
much less tightly bound to the protein than, for example, 
flavine adenine dinucleotide, perhaps to facilitate move- 
ment of the available supply among the apoenzymes in 
need of it. 

Some of the many microbial reactions known to require 
diphosphopyridine nucleotide (DPN) or triphosphopyri- 
dine nucleotide (TPN) are: 

1 Joan F. Powell, Biochem. ]. 54 210 (1953). 

-J. J. Perry and J. W. Foster, J. Bacteriol. 72 295 (1956). 

■^ William K. Harrell and Emil Mantini, Can. J. Microbiol. 3 735 
(1957). 

^ Joan F. Powell and R. E. Strange, Biochem. J. 65 700 (1957). 



Pfizer Handbook of Microbial Metabolites 480 

CH3CH2OH ;=i CH3CHO (in yeast) 
R— CHO ^ R— COOH (in yeast) 
Glutathione — SH ;^ Glutathione — S — S — Glutathione (in yeast) 
Isocitrate :?^ Oxalosuccinate (bacteria, yeast) 
D-Glucopyranose-6-phosphate :?^ 6-Phospho-D-gluconolactone (yeast) 

L-GIutamate ;=i a-Ketoglutarate + NH4 (bacteria) 
D-Glyceraldehyde-3-phosphate + Phosphate ;=i D-l,3-Diphosphoglyceric 

Acid (yeast) 

Some of these reactions occur quite generally. Oc- 
casionally DPN and TPN are interchangeable, although 
one or the other is used more efficiently. 

Direct transfer of hydrogen between the substrate and 
the 4-position of the nicotinamide moiety of DPN (in the 
presence of yeast alcohol dehydrogenase) has been dem- 
onstrated, and the stereochemistry of this reaction studied 
in exquisite detail by means of deuterated substrate : ^ ® 



CONH2 


H D 
\/ CONH2 


fY 


+ CH3CD2OH ^ 11 -f CH3CDO + H® 


1 ® 


1 


R 


R 




(R = the rest of the DPN molecule) 


"w" CONH. 


CONH. H 

/ 1 


(ill + CH3— CHO + H ® ;=i (T^Y + CH3— C— OH 


1 


1© D 


1 

R 


R 



In the second equation the deuterium atom is removed 
exclusively, leaving deuterium-free DPN. This indicates 
a marked- steric effect, since the deuterium atom projects 
from one side of the molecule. Moreover, a single stereo- 
isomer of deuterated ethanol is produced. 

Speculations have been made concerning the precise 
nature of the coenzyme-apoenzyme-substrate-metal ion 
complex. One model' is shown below: 

^ Harvey F. Fisher, Eric E. Conn, Birgit Vennesland and F. H. 
Westheimer, J. Biol. Chem. 202 687 (1953). 

^ H. Richard Levy, Frank A. Loewus and Birgit Vennesland, J. Am. 
Chem. Soc. 79 2949 (1957). 

^Kurt Wallenfels and Horst Sund, Biochem. Z. 329 59 (1957). 



48i 



Pyridines 



0*-P— OH 




0<-P— OH 



The fact that alcohol and lactic acid dehydrogenases all 
have been found to contain 2 or 4 DPN molecules has also 
inspired the hypothesis that hydrogen transfer might re- 
quire a pair of adjoining prosthetic groups in a scheme 
such as : 






in which a deuterated substrate is shown for clarity.* A 
more detailed discussion has been pubUshed of the stereo- 

^ Jan van Eys, Anthony San Pietro and Nathan O. Kaplan, Science 
127 1443 (1958). 



Pfizer Handbook o£ Microbial Metabolites 



482 



chemistry of microbiological reactions with emphasis on 
those promoted by dehydrogenases. ° 

The biosynthesis of nicotinic acid has been studied in 
several different biological systems. In neurospora (and 
in mammals) tryptophan is the source with 3-oxyanthra- 
nilic acid a proved intermediate.^" "• ^^' " ^*' ^^ The re- 
maining stages of this route are obscure, although a- 
aminomethyl-a,/i^-trarzs-y,8-cis-muconic acid may be an in- 
termediate.^'^ It has been shown to be a precursor of 
nicotinic acid for the bacterium Xanthomonas pruni. If 
it proves to be generally significant, then the following 
scheme can be written: 

O NH2 



CH, 




-CH— COOH 
NHo — 



~7^ 

o, 



Tryptophan 



NH2 



C— CH,— CH— COOH 

CHO 
^NH-^^ 
N-Formylkynurenine 

NH2 



CO: 
[O] 




CO— CH,— CH— COOH 



NHo 




Kynurenine 



CO— CHj— CH— COOH 



NH. 



3- Hydroxy kynurenine 



»G. E. W. Wolstenholme and Cecilia M. O'Connor (Eds.), CIBA 
Foundation Study Group No. 2, "Sterlc Course of Microbiologi- 
cal Reactions," Little, Brown and Company, Boston, 1959, 115 pp. 

" W. A. Krehl, L. J. Teply, P. S. Sarma and C. A. Elvehjem, Science 
101 489 (1945). 

" Fred Rosen, Jesse W. Huff and William A. Perlzweig, /. Biol. 
Chem. 163 343 (1946). 

'- G. S. Beadle, H. K. Mitchell and J. F. Nye, Proc. Nat. Acad. Sci. 
33 155 (1947). 

'^ Francis A. Raskins and Herschel K. Mitchell, ibid. 35 500 
(1949). 

1* Irving L. Miller and Edward A. Adelberg, /. Biol. Chem. 205 691 
(1953). 

I'^WiUiam B. Jakoby and David M. Bonner, ibid. 205 699, 709 
(1953). 

"J. O. Harris and F. Binns, Nature 179 475 (1957). 



483 



Pyridines 





COOh 
HO .^^^ 

V M if 


\ 


COOH" 


H,0 


CH;r 


-cIh— COOH WA 

1 1 NH, 
NH, OH 


1 ^NH.. 
_ 




3-Hydroxy- Keto-form 
anthranilic Acid 






H 
C 

/\ 
HC CH CO, 

II 1 -^ 
HC CH — NH, 


r COOH" 










H, 




\ 

COOH 


_ H 








a-Aminomethyl-a, 

0-trans-y, 5-cis- 

muconic Acid 


T( 
ni 


jtrahydro- 
:otinic Acid 







COOH' 




^N 
H 

Dihydro- 
nicotinic Acid 



COOH 



H2 




Nicotinic 
Acid 



A different method of biosynthesis exists in Escherichia 
coli and Bacillus suhtilis since tryptophan is not used. In- 
vestigation of this route has not progressed so far, but 
glycerol is capable of supplying all carbon atoms, as are 
glyceric acid and dihydroxy acetone (but not pyruvate). 
Succinate, malate, fumarate and oxaloacetate also were 
used. Ribose and adenine were required, which suggests 
direct synthesis of the coenzyme. ^^ 



^' Manuel V. Ortega and Gene M. Brown, /. Am. Chem. Soc. 81 
4437 (1959). 



Pfizer Handbook of Microbial Metabolites 484 





The various forms of pyridoxine 


are: 


CH20H 

"O. 1 /CH2OH 


HO f 
CH3 


HO CH2NH2 

CH2OH j^Q^j. HO 1 CH2OH 

5^/ amination r^l 


CH3 




if^ 


CH3 


Pyridoxine 




Pyridoxal 


Pyridoxamine 


• If 

CH2OH 
"O 1 CH2OPO3H2 

1 


HO y"° CH2OPO3H2 


^^ CH2NH2 

HO 1 CH2OPO3H2 


CH3 




CH3 


CH3 


Pyridoxine Phosphate 


Pyridoxal Phosphate 


Pyridoxamine Phosphate 



Virtually nothing is known concerning the biogenesis of 
pyridoxine. Since catabohsm often furnishes clues useful 
in the study of biosynthesis, it should be noted that oxida- 
tive bacteria degrade pyridoxine as follows : ^* 



CH2OH 



CH2OH CH2OH 



HO I ' CH2OH HO I CHO HO COOH 



CH3' CH3 CH3 

Pyridoxine Isopyridoxal 5-Pyridoxic Acid 

(Pyridoxol) (Lactone) 

CH2-O 



HO ' ^ 



O 



CH3 

i 



18 Victor W. Rodwell, Benjamin E. Volcani, Miyoshi Ikawa and 
Esmond E. Snell, /. Biol. Chem. 233 1548 (1958); Miyoshi Ikawa, 
Victor W. RodweU and Esmond E. SneU, ibid. 233 1555 (1958). 



485 Pyridines 



i 
CH2OH 

I 

HOOC C 

o II 



COOH 



CH 



CH3 



^NH" 



a-Hydroxymethyl-a'- 

(N-acetylaminomethylene)- 

succinic Acid 



Acid hydrolysis converts the acychc product to paraconic 
acid. 

Functions of the vitamin are better understood. The 
names pyridoxine or vitamin B^ commonly are used in a 
general sense to refer to the group. Pyridoxal 5-phosphate 
is the actual prosthetic group in most enzymic reactions. 
It is a component of transaminases, amino acid decar- 
boxylases, tryptophan synthetase, amino acid racemases, 
threonine synthetase (homoserine isomerase), S-amino- 
levulinate synthetase, phosphorylase and various other 
enzymes which manipulate amino acids. More thorough 
discussions of functions of this important vitamin can be 
found in reviews. ^^' ^° 

Some pyridoxal-catalyzed reactions can be carried out in 
aqueous solution without the apoenzymes if heat and the 
proper metal ions (Al^"^*, Fe**, Cu++) are supplied. Mech- 
anisms which have been proposed for three such reactions 
are outhned in the following equations:-^' ^-- ^^- ^* 



^^ Esmond E. Snell, Vitamins and Hormones 16 77 (1958). 

-"Paul D. Boyer, Henry Lardy and Karl Myrback, (Eds.) "The 
Enzymes," Alexander E. Braunstein, Pyridoxal phosphate, Academic 
Press, New York, 1960, pp. 113-184. 

-^ David E. Metzler, Mlyoshi Ikawa and Esmond E. Snell, /. Am,. 
Chem. Soc. 76 648 (1954). 

"J. B. Longenecker and Esmond E. Snell, ibid. 79 142 (1957). 

23 W. Terry Jenkins and Irwin W. Sizer, ibid. 79 2655 (1957). 

-* D. S. Hoare and Esmond E. Snell, Proc. Internat. Sympos. Enz. 
Chem., Tokyo and Kyoto, Pergamon Press, London, 1957, p. 142. 



Pfizer Handbook of Microbial Metabolites 



486 



HOCH; 



CHO 




OH 

4- CH,— COOH 
CH3 NH, 



® \X 2H® 



HC C--0 

II i 

HOCHo HC M 

J' .0 




N' 

H CH3 

CH2O 

H© 

NH2 
HOCH2— CH— COOH 

+ HOCH2— C C 

CHO _.. H' 



H 



HOCH2 




OH ^N^ jO 

H.,o HOCH2 HC M 



HOCHo— C— COOH 

+ 

NHo 



CH. 
HOCH2 I " OH 




2H© 
H2O 



CH3 



i® 



CH3 




HOCH — C CO 

li 1 

HOCH. HC M 
.0 




H"^ CH3 



N' 

H CH3 



CH3— C— COOH 

+ 
NHa 



H.O 



NH2 
CH2=C— COOH. 

+ 



CHO 
HOCH2 I OH 2H® 




H2O 



CH3 



OH© 



H2C-=C C=-0 

HOCH. HC M 
,0 




H® CH3 



M = Metal: (T),(2) = Aldol formation and cleavage 
Ql® — Transamination 
(3), (5) = «. d-Elimination 



487 Pyridines 

Attachment to the apoenzyme in vivo was assumed to 
be at the pyridine nitrogen atom. Spectral data from such 
model systems, however, when applied to purified en- 
zymes, indicate that pyridoxal phosphate is bound to the 
apoenzyme as a Schiff base in glutamate-aspartate amino- 
pherase'-'* and in homoserine deaminase-cystathionase.^^ 
In crystalline muscle phosphorylase pyridoxal is bound to 
the apoenzyme, probably at a lysine e-amino group, as an 
aldamine, involving an additional side-chain of the protein 
(perhaps — SH).-'' -' 

Protein 

Protein HN X ^ 

o© \ / o® 



,N^ XH 



CH. 



H' ^CH (-H_o_p_OH ®0 I ' CHo— O— P— OH NaBH4 




o 

CH3 CH3 H® 




O 



Schiff Base Aldamine (X = S?) 

Protein 



HN XH 



O© 



CH I 

©O I ' CH.— O— P— OH 

i 

o 




Reduced Enzyme 

Glutamate-aspartate aminopherase contains 2 moles of 
bound pyridoxal phosphate and muscle phosphorylase 4. 
It is rather surprising to find the vitamin in an enzyme, 
such as the latter, unrelated to its ordinary function. 
Doubt has been cast on its function as a prosthetic group 
in phosphorylase by several experiments, one of them the 
reduction shown, which should have inactivated the 
pyridoxal, but which did not inactivate the enzyme.-'^ It 

-^ Yoshihiko Matsuo and David M. Greenberg, /. Biol. Chem. 230 
545, 561 (1958); idem., ibid. 234 507, 516 (1959). 

-° Alan B. Kent, Edwin G. Krebs and Edmond H. Fischer, /. Biol. 
Chem. 232 549 (1958). 

-" Barbara Illingworth, Hendrlk S. Jansz, David H. Brown and 
Carl F. Cori, Proc. Nat. Acad. Sci. 44 1180 (1958). 

■-** Edmond H. Fischer, Alan B. Kent, Eloise R. Snyder and Edwin G. 
Krebs, /. Am. Chem. Soc. 80 2906 (1958). 



Pfizer Handbook of Microbial Metabolites 488 

may be that it serves a structural or other function here. 

D-Cycloserine has been reported to inhibit aspartate 
aminopherase, indole synthetase and D-alanine-D-gluta- 
mate aminopherase in some bacteria.-^' ^° Aspartic ana- 
logues, such as diaminosuccinic acid and hydroxyaspartic 
acid also are effective inhibitors of the first enzyme 
above. ^^ 

It has been suggested that pyridoxine may be impli- 
cated in the active transport of amino acids across cell 
walls. ^^ 

968 2, 6-Dipicolinic Acid, C7H5O4N, colorless needles, m.p. 236° 

(dec). 




HOOC COOH 

Bacillus megatherium, B. cereus var. terminalis, B. 
sphaericus types 

Occurs as the calcuim salt in spores. 

Joan F. Powell, Biochem. J. 54 210 (1953). 

William K. Harrell and Emil Mantini, Can. J. Microbiol. 3 
735 (1957). 

Joan F. Powell and R. E. Strange, Biochem. J. 65 700 
(1957). 



969 Pyridoxal-5'-phosphate CgHioOgNF 

O 



OH 



HO I CH2— O— P=0 



OH 



>N 



CH3 
Yeasts, molds, bacteria (widely distributed) 

29Takakazu Aoki, Kekkaku 32 544, 605 (1957). (Chem. Ahstr. 52 
7427g). 

30 N. K. Kochetkov, R. M. Khomutov, M. J. Karpeiskii, E. I. Budov- 
skii and E. S. Severin, Doklady Akad. Nauk S.S.S.R. 126 1132 (1959). 

3^ Mario Garcia-Hernandez and Ernest Kun, Biochim. et Biophys. 
Acta 24 78 (1957). 

3- Halvor N. Christensen, Thomas R. Riggs and Barbara R. Coyne, 
J. Biol. Chem. 209 413 (1954); Halvor N. Christensen and Thomas R. 
Riggs, ibid. 220 265 (1956). 



489 Pyridines 

I. C. Gunsalus, W. D. Bellamy and W. W. Umbreit, /. Biol. 
Chem. 155 685 (1944). 

Dorothea Heyl, Eileen Luz, Stanton A. Harris and Karl 
Folkers, ;. Am. Chem. Soc. 73 3430 (1951). (Synthesis) 

970 Pyridoxine (Vitamin B,j), CsHuOyN, colorless needles from 
acetone, m.p. 160° (sublimes). 

CH.OH 
HO I CH2OH 



CH3 

Yeasts, molds. 

Yields of 82-114 ^g. per gram (dry basis) have been 
reported from penicillin broth filtrates. 

Yields of 23-100 ^g. per gram of dry cells have been 
reported from brewers' yeast. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
1954 Vol. II, J. M. VanLanen, Production of vitamins other 
than riboflavin, Chap. 6, pp. 191-216. (A review) 

971 Ethyl Hydrogen 2, 6-DipicoIinate, C9H9O4N, colorless crystals, 
m.p. 121.5°. 




HOOC COOC2H5 



Bacillus cereus var. mycoides (spores) 

J. J. Perry and J. W. Foster, J. Bacteriol. 72 295 (1956). 

972 Dehydrofusaric Acid, C10H11O2N, colorless crystals, m.p. 118° 

CH2=<:H— CH2— CH2 



COOH 



Gibberella fujikuroi Saw. 

Ernst Gaumann, Phytopathology 47 342 (1957). 
C. A. Stoll and J. Renz, Phytopathol. Z. 27 380 (1957). 
John Frederick Grove, P. W. Jeffs and T. P. C. Mulholland, 
J. Chem. Soc, 1236 (1958). 




Pfizer Handbook of Microbial Metabolites 



490 



973 Fusaric Acid, C10H13O2N, colorless crystals, m.p. 100°. 



CH3CH2CH2CH0 



COOH 




Gibberella fujikuroi (Saw.) Wr., Fusarium heterospo- 
rum Nees, F. bulbigenum Cke. et Mass. var. lycopersici 
(Bruchi) Wr. et Rg., F. vasinfectum Atk., F. orthoceras 
App. et Wr., Nectria cinnabarina (Tode) Fr. 

Yields of about 0.5 g. per liter have been reported. 

Teijiro Yabuta, Katsuji Kambe and Takeshi Hayashi, /. Agr. 
Chem. Soc. Japan 10 1059 (1934). 

John Frederick Grove, P. W. Jeffs and T. P. C. MulhoUand, 
J. Chem. Soc, 1236 (1958). 

974 Coenzyme III (Nicotinamide Ribose 5'-Diphosphate), CnHieOn- 

NoPo. 



O 

II 

C— NH2 



o© o 

I II 

0=P— O— P— O— CHo 




& 



OH 



OH 




OH OH 



Yeast 

Nicotinic acid nucleotides also have been isolated from 
yeast. 

Thomas P. Singer and Edna B. Kearney, Biochim. et 
Biophys. Acta 11 290 (1953). 



491 



Pyridines 



975 Diphosphopyridinenuclcotide (DPN), C2iH^,70,4N7P^.. 
NH, 

c.r.> 




OH OH 



Yeasts, molds (widely distributed) 

H. von Euler, P. Karrer and B. Brecker, Helv. Chim. Acta 
19 1060 (1936). (Structure) 

G. A. LePage, /. Biol. Chem. 168 623 (1947). 

976 Triphosphopyridinenucleotide (TPN, Codehydrase II), C.>iH<,g- 
OitNtPs. 

NH, 




OH 



O© 



^V CHo— O— P— O— P— O— CM 

oh\ II II 

^^ O o 




OH OH 



Yeasts, molds, etc. 

Otto Warburg, Walter Christian and Alfred Griese, 
Biochem. Z. 279 143 (1935); 282 157 (1935). (Isolation) 



Pfizer Handbook of Microbial Metabolites 



492 



H. von Euler and F. Schlenk, Z. physiol. Chem. 246 64 
(1937). (Structure) 

Arthur Kornberg and W. E. Pricer, Jr., /. Biol. Chem. 186 
557 (1950). 



;. QUINOLINES 



Quinolines are produced by bacteria and molds, but ap- 
parently none has been reported from streptomycetes or 
lichens. A complex of seven related 4-oxyquinolines is 
elaborated by the oxidative bacterium Pseudomonas aeru- 
ginosa (Bacillus pyocyaneus). These are commonly 
called "pyo" compounds. 

Evidently no investigations have been made on the 
mode of biosynthesis of microbial quinolines. The isola- 
tion of anthranilic acid and of 2-n-heptyl-3-oxy-4-quino- 
lone from "pyo" fermentation broths is suggestive, how- 
ever.^ It seems probable that the "pyo" compounds could 



COOH 



NH2 
Anthranilic 
Acid 



OH 



N' 

H CH2CH2CH2CH2CH2CH2CH3 
2-n-Heptyl-3-oxy-4-quinolone 



be formed essentially by condensation of anthranilic acid 
or a biosynthetic precursor with a fatty acid or a fatty 
acid precursor: 



O 

II 
C— OH 



COOH 




+ 



NH2 



H2C 

c 

/\ 

O R 

O 



/ CH 



COOH 



.C— OH 



1. Oxidative 
decarboxylation 

2. Dehydration 



OH 



IH] 





iRokuro Takeda, J. fermentation Technol. 37 59 (1959). 



493 



Quinolines 



Oxidative decarboxylation would then yield an inter- 
mediate of the type isolated, and a one-stage reduction the 
4-oxyquinolines. The N-oxides might be formed later by 
post-oxidation. Quinolines are known to be quite suscep- 
tible to N-oxidation by peroxides or oxygen. 

The structure of the mold product, viridicatin, has been 
verified by synthesis, while that of cyclopenin is still un- 
certain. It would appear that these substances are also 
derivatives of anthranilic acid. In this case, condensa- 
tion probably occurs with an earlier member of the shiki- 
mic acid pathway, perhaps prephenic acid or phenyl- 
pyruvic acid: 



H2C 

COOH c=0 



-CO2 




NH, 



Anthranilic 
Acid 



c 

O OH 



CH 

y \ 

/ c=o 



\ 



// 



OH 




Phenylpyruvic 
Acid 



Viridicatin 



Such condensations have been suggested to explain the 
origin of certain oxyquinoUne plant alkaloids. - 

There is, of course, a possibility for 4-oxyquinoLine for- 
mation by way of tryptophan and kynurenine : 




O 

C 

\ 
CH. 

1 
NH2 CH— NH2 



transamination, 
dehydration 




COOH 



COOH 



Kynurenine 

This seems to be an unnecessarily indirect route, but all 
of the schemes shown here await experimental test. 

Ernest Wenkert, Experientia 15 165 (1959). 



Pfizer Handbook of Microbial Metabolites 494 

977 Viridicatin, Ci-.H^O^N, colorless needles, m.p. 268°. 




Penicillium viridicatum Westling, P. cyclopium West- 
ling 

See under cyclopenin. 

A. Bracken, Anna Pocker and H. Raistrick, Biochem. J. 57 
587 (1954). (Synthesis) 

978 2-n-Heptyl-4-oxyquinoline, CjcHoiON, colorless crystals, m.p. 
146°. 



OH 




CH2CH2CH2CH2CH.CH2CH3 

Pseudoinonas aeruginosa 

These quinoline derivatives are called "pyo" compounds. 

Ibert C. Wells, /. Biol. Chem. 196 331 (1952). (Synthesis) 

979 2-n-HeptyI-3-oxy-4-quinolone, CieH.iO.N. 

O 




H CH,CH2CH.CH2CHnXH2CH3 

Pseudomonas aeruginosa strain T-359 

The other "pyo" compounds were isolated as well as 
anthranilic acid, pyoluteorin, pyocyanine, phenazine-1- 
carboxylic acid and oxychlororaphine. 

Rokuro Takeda, /. Fermentation Technol. 37 59 (1959). 



495 Quinolines 

980 2-n-Heptyl-4-oxyquinoline N-oxide, CicH^iOoN, colorless leaflets, 
m.p. 158-160°. 




i CH,CH,CH,CH2CH,CH,CH3 
O 

Pseudomonas aeruginosa 

J. W. Cornforth and A. T. James, Biochem. J. 63 124 
(1956). (Synthesis) 

981 Cyclopenin, CiyHj^O^N., colorless tablets, m.p. 207°, [a],r" 
-306° (c 1.0 in ethanol). 
Proposed structures : 



OH 

N— CHs 







H OH 



Penicillhnn cyclopium Westling 

Usually viridicatin is produced by the same organism. 
Palitantin and frequentin are also produced by P. cyclo- 
pium 

A. Bracken, Anna Pocker and H. Raistrick, Biochem. J. 57 
587 (1954). 

982 2-(n-A'-Nonenyl)-4-oxyquinoline, Ci,t;Ho;,ON, colorless crystals, 
m.p. 153°. 




CH=CHCH2CH2CH2CH2CH2CH2CH3 

Pseudomonas aeruginosa 

Ibert C. Wells, /. Biol. Chem. 196 331 (1952). (Synthesis) 



Pfizer Handbook of Microbial Metabolites 496 

983 2-n-Nonyl-4-oxyquinoline, CigHogON, colorless crystals, m.p. 
139°. 




CH2CH2CH2CH0CH0CH2CH2CH2CH3 

Pseudomonas aeruginosa 

Ibert C. WeUs, J. Biol. Chem. 196 331 (1952). (Synthesis) 

984 2-n-Nonyl-4-oxyquinoline N-Oxide, C18H25O2N, colorless leaflets, 
m.p. 148°. 

OH 




i CHzCHsCHzCHzCHjCHzCHzCHiCHj 
O 

Pseudomonas aeruginosa 

J. W. Cornforth and A. T. James, Biochem. J. 63 124 
(1956). (Synthesis) 

985 2-n-Undecyl-4-oxyquinoline N-Oxide, C20H09O2N, colorless leaf- 
lets, m.p. 148.5°. 




i CH2CH2CH2CH2CH2CH2CH2CH2CH2CH2CH3 
O 

Pseudomonas aeruginosa 

J. W. Cornforth and A. T. James, Biochem. }. 63 124 (1956). 
(Synthesis) 

k. PYRAZINES, DIKETOPIPERAZINES 

Diketopiperazines are produced by molds, yeasts and 
lichens, but none has been reported from bacteria. Be- 
sides those listed in this section, others are classified else- 
where, for example, echinulin and gliotoxin under indoles. 

Flavacol and pulcherriminic acid seem to be derived 



497 



Pyrazines, Diketopiperazines 



from leucine, the echinulin moiety from leucine and ala- 
nine, aspergillic acid from leucine and isoleucine, the 
mvcelianamide moiety from tyrosine and alanine, picroro- 
cellin from phenylalanine, and gliotoxin from phenyl- 
alanine and serine. It might be mentioned that we have 
isolated from a Rhizopus nigricans culture a diketopipera- 
zine which is a derivative of isoleucine and valine (un- 
published). 

Dehydration, dehydrogenation, oxidation and N- or 
O-methylation sometimes occur to obscure the origin to 
some degree. Aromatization to a pyrazine has taken place 
in flavacol and pulcherriminic acid, aspergillic acid, a 
dihydropyrazine, representing an intermediate oxidation 
state. Formation of pulcherriminic acid might be repre- 
sented as follows: 



OH H2N 



CH3 



CH3 



\ 



CH— CH2— CH 

I \ 

CH— CH2— CH C= O CH3 

/ \ / 

CH3 NH2 HO 

H 



-H2O 



CH3 



CH3 



\ 



O [\ CH3 

CH3 C CH— CH2— CH H2O 

\ II \ — 

CH— CH2— CH C=0 CH3 

/ \ \ 

CH3 NH2 OH 

CH3 "— 



CH— CH2— CH 



CH— CH2— CH C=0 
H 



CH3 



CH3 



CH3 



\ 



HO— C C— CH2— CH — H2 

CH3 



CH— CH2 



C C— OH 

H 



CH3 



Pfizer Handbook of Microbial Metabolites 498 



CH3 



HO CH.— CH O. 

CH3 f i CH3 

CH— CH2 OH 



CH3 



CH3 
O / 

HO T CH2— CH 

CH3 'I I ^^^ 



K^ 



CH— CH, i OH 
/ O 

CH3 

The addition of sulfur across the diketopiperazine ring 
in gliotoxin is interesting. 

986 Flavacol, CjoH^oONo, colorless needles, m.p. 147-149°. 

CH3 

/ 
CH2— CH 



K 



CH3 II J C"' 



CH— CH 2 Oh 

/ 
CH3 

Aspergillus flavus 

George Dunn, G. T. Newbold and F. S. Spring, /. Chem. Soc. 
2586 (1949). (Synthesis) 

987 Aspergillic Acijcl, CjoHooOoN^, pale yellow needles, m.p. 97-99°, 
[a]D-" +13.4° (c 1 in ethanol). 

CHs 

/ 
H CH2— CH 

CH3 



CH3— CH. 

CH i OH 
/ O 

CH3 



Aspergillus flavus 



499 Pyrazines, Diketopiperazines 

James D. Butcher, /. Biol. Chem. 232 785 (1958). 

988 Granegillin, C,oHo„OoNo, pale yellow needles, m.p. 99-100°, 

optically inactive, the crystals have a characteristic odor 
(as does Aspergillic Acid). 

The only important difference in properties between 
granegillin and aspergillic acid is the lack of optical activ- 
ity in the former, and the two compounds may be identical. 

A mold resembling Aspergillus flavus 

A. Csillag, Acta Microbiol. (Hungary) 1 321 (1954); Abstr. 
in Bull. Hijg. 30 159 (1955). 

989 Hydroxyaspergillic Acid, Ci2H2„0;iN2, nearly colorless needles, 

m.p. 148-150°, [a],r' +36° (c 1 in ethanol). 

CH3 

/ 
CH.— CH 

CH3 I ^"•■' 

CH:, CHo— CH \ OH 
I O 

OH 

Aspergillus flavus 

James D. Butcher, J. Biol. Chem. 232 785 (1958). 

990 Neohydroxyaspergillic Acid, C10H20O3N2, colorless crystals, m.p. 

164-166°, [alo'' -58° (c l".01 in ethanol). 

Aspergillus sclerotiorum 

A yield of about 300 mg. per liter was obtained. 

Ulrich Weiss, Frieda Strelitz, Helen Flon and Igor N. Ashe- 
shov, Arch. Biochem. and Biophys. 74 150 (1958). 

991 



iminic Acid, 


Cx 


2H20O4N0, 


m.p. 


173° 


CH3 












\ 













CH- 


-CH2 


T 


OH 




/ 
CH3 




\ 


.N^ 


< 








/ 


-N^'\ 








HO 


i 



CH2 


— CH 



CH3 



CHa 



Candida pulcherrima (Lindner) Windisch 

This compound was isolated as a red, iron-complexed 



Pfizer Handbook of Microbial Metabolites 500 

pigment called pulcherrimin, which probably has the 
structure : 

CH3 III 

\ O - Fe 

CH— CH2 T 1 

CHa f J CHa 

I III i CH2 — CH 
Fe - O \ 

CHs 

The yield was 30 mg. of pulcherrimin per gram of dry 
cells. 

A. J. Kluyver, J. P. van der Walt and A. J. van Triet, Proc. 
liat. kcad. Sci. U. S. 39 583 (1953). 

A. H. Cook and C. A. Slater, J. Chem. Soc, 4130, 4133, 
(1956). (Structure) 

992 Picrorocellin, C20H22O4N2, colorless prisms, m.p. 192-194°, [ajn 

+ 12.5°. 

CH3 O 

\ / 
N-C 

CH— CH CH— CH- 

I Vnh I 

OCH3/ OH 

O 

Roccella fuciformis Ach. 

Martin Onslow Forster and William Bristow Saville, 
J. Chem. Soc, 816 (1922). 

993 Mycelianamide, CooHogOgNs, colorless leaflets, m.p. 170-172° 

(dec), [a]546i'' -217° (c 0.869 in chloroform). 

HO O 

\ / 

O^N-C 
-CH=C CH— CH3 

c-n' 

O OH 

Penicillium griseofulvum 



I 



50I Phenazines and Phenoxazones 

A. J. Birch, R. A. Massy-Westropp and R. W. Rickards, 

;. Chem. Soc, 3717 (1956). 

A. J. Birch, Proc. Chem. Soc, 233 (1957). 



/. PHENAZINES AND PHENOXAZONES 

The phenazine bacterial pigments have been known for 
many years. Pyocyanine was probably isolated in the 
early 1860's, and oxychlororaphine was synthesized in 
1930. New pigments of this type continue to be reported, 
usually from pseudomonas species, but also from strepto- 
mycetes. Pyocyanine is responsible for the blue-green 
color of pus, since Pseudomonas aeruginosa is a skin para- 
site, and certain other blue or green stains on natural ma- 
terials have been identified with phenazine pigments. 

The phenazine bacterial pigments have been reviewed,^ 
and this introduction will be confined to a few remarks on 
biosynthesis. Actually, there is as yet little to be said on 
this subject. Several studies have been made concerning 
medium requirements and improvements for optimum 
pigment production in both growing- and stationary cul- 
tures.* In growing cultures a yield of 231 mg. of pyocya- 
nine per liter was obtained on a medium containing glyc- 
erol, D,L-alanine, L-leucine and magnesium, calcium, phos- 
phate, sulfate and ammonium ions. 

In resting cultures glutamic acid and y-aminobutyric 
acid were found to be the best substrates, yielding about 
250 mg. of pyocyanine per liter. Pigment production was 
slow and inhibited by respiratory poisons (cyanide, azide) 
but not by fluoride. 

These results are not very helpful in speculations on 
the biosynthetic intermediates. 

Viewed in aggregate there is a noticeable recurrence of 
either hydroxyl or carboxyl groups at the 1-position, the 
9-position or the 6-position of the phenazine nucleus. 

^ G. A. Swan and D. G. I. Felton, "Phenazines," Interscience Pub- 
lishers, Inc., New York, 1957, pp. 174-191. 

- M. O. Burton, J. J. R. Campbell and B. A. Eagles, Can. J. Res. 
26C 15 (1948); M. O. Burton, B. A. Eagles and J. J. R. Campbell, 
ibid. 25C 121 (1947); G. Young, /. Bacteriol. 54 109 (1947); Esther 
HeUinger, J. Gen. Microbiol. 5 633 (1951). 

^ N. Grossowicz, Peyuta Hayat and Y. S. Halpern, /. Gen. Microbiol. 
16 576 (1957). 



Pfizer Handbook of Microbial Metabolites 502 

This is reminiscent of the phenoxazones such as cinna- 

HOCH, COOH OCH3 COOH 

I /NH2 I 





HO— CH2— C— O— CH2 
Phenazine Cinnabarin Griseolutein A 

barin and actinocinin. The analogy perhaps can be de- 
veloped farther, since a streptomycete pigment has been 
found with an amino group in the 2-position. 

The resemblance is sufficient to suggest anthranilic acid 
or related substances as intermediates in the biosynthesis 
of phenazines. Oxidative decarboxylations of aromatic 
acids to phenols are not unknown among obligate aerobes 
of the type that produce phenazines. Also 3-oxyanthra- 
nilic acid might account for some of the phenolic hydroxyl 
groups. 

As for the coupling reaction, perhaps something akin 
to phenolic-free radical coupling takes place. Photoirra- 
diation of aniline at low temperatures has been reported 
to produce phenazine.* Also tetraphenylhydrazine heated 
to 90° apparently dissociates to a free radical which re- 
arranges to (among other things) a dihydrophenazine.^ 

NHo 

light ^;;?=\^N^ 



low temperature ^-^'^^N 



// \ ^ // \ // % 



_/ Heat \_/\... r^^^^^f^^^ 



N— N ^ N- 

// \\/ \r\ r\^ 



'A. N. Terenin, Acta Physicochim. S.S.S.R. 1.-5 1 (1940); Chem. 
Abstr. 35 1701 (1941). 

■'"' G. W. Wheland, "Advanced Organic Chemistry," John Wiley 
and Sons, Inc., New York, 1949, pp. 727-728. 



503 Phenazines and Phenoxazqnes 

Atmospheric oxidation is enough to cause phenazine 
formation from 3,4-diaminoguaiacol.'' This is a favor- 
able case for free radical stabilization. 

^^"^NH.- HO ?^"^ ?^"^NH. 

\ ^ .M^ O. 




NHo NH, NH2 




This argument of course is speculative. 

994 1-Phenazinol (1-Hydroxyphenazine, Hemipyocyanine ) , Cj^Hg- 
ON2, orange crystals, m.p. 157° (sublimes). 



OH 






Pseudomonas aeruginosa 

Fritz Wrede and E. Strack, Z. physiol. Chem. 177 177 
(1928). 

G. M. Badger, R. S. Pearce and R. Pettit, /. Chem. Soc. 
3204 (1951). 

Walter S. Moos and John W. Rowen, Arch. Biochem. and 
Biophijs. 43 88 (1953). 

995 1,6-Dihydroxyphenazine, CioH^OmN:,, golden yellow prisms, m.p. 
274°. 

OH 

I 
HO 

Streptomyces thioluteus 

Hideshi Akabori and Michikazu Nakamura, /. Antibiotics 
(Japan) 12A 17 (1959). 

<^Fr. Fichter and Julius Schwab, Ber. 39 3339 (1906). 



Pfizer Handbook of Microbial Metabolites 504 



996 lodinin (l,6-Phenazmediol-5,10-dioxide), C12H8O4N2, purple 
crystals with a coppery glint, m.p. 236° (dec.)- 



? OH 

T I 



HO 



i 
O 



Chromobacterium iodinum 

G. R. Clemo and A. F. Daglish, J. Chem. Soc, 1481 (1950). 

997 Phenazine-1-carboxylic Acid, CigH^O^N^, greenish yeUow needles, 
m.p. 242°. 

COOH 



Pseudomonas aureofaciens Kluyver, Streptomyces misa- 
kiensis, Calonectria 

Yields of about 1 g. per liter have been mentioned. The 
streptomycete produced another phenazine, C17H16N0O2, 
called tubermycin A. A pigment closely related to phena- 
zine- 1-carboxylic acid was also isolated by Kluyver from 
the pseudomonas organism. 

A. J. Kluyver, J. Bacteriol. 72 406 (1956). 

Wm. C. Haynes, Frank H. Stodola, Joan M. Locke, 
Thomas G. Pridham, Howard F. Conway, Virgil E. Sohns and 
Richard W. Jackson, ibid. 72 412 (1956). 

Kiyoshi Isono, Kentaro Anzai and Saburo Suzuki, /. Anti- 
biotics (Japan) llA 264 (1959). 

998 Oxychlororaphine, C13H9ON3, yellow needles, m.p. 237° (sub- 
limes in" the absence of O^, giving yellow crystals, m.p. 
241°). 

CONH2 




Pseudomonas chlororaphis 

Fritz Kogl and J. J. Postowsky, Ann. 480 280 ( 1930). (Syn- 
thesis ) 



505 Phenazines and Phenoxazones 

999 Chlororaphine, green crystals, m.p. (in the absence of Oo) 225° 
(dec.) (in the presence of Oo sublimes at 210°, giving a 
yellow sublimate). 

Chlororaphine in the crystalline state is a molecular 
compound of phenazine-1-carboxamide and its dihydro 
derivative in the ratio of 3 : 1. 

CONHo CONHo 

:xb - cc 

H 

Charles Dufraisse, Andre Etienne and Edmond Toromanoff, 
Compt. rend. 235 920 (1952). 

But in solution in the pH range 1.75-10.85 (particularly 
at lower pH) chlororaphine exists largely in the semi- 
quinone form: 

CONH2 



CONH2 

H 1 




HO 


/N\^^ <- 


^f^ 


^/Nx. 


© i J 


k/ 


!l © I 


H© 




H 



Carlo Cattaneo, Guido Sartori and M. Morellinl, Gazz. chim. 
ital. 77 381 (1947). 

Pseudomonas chlororaphis 

Fritz Kogl and J. J. Postowsky, Ann. 480 280 (1930). 

1000 Pyocyanine, CjgHisNoO, dark blue needles, m.p. 133°, decom- 
poses to 1-phenazinol on standing in light and air. 

O© 



I® 
CHs 

Pseudomonas aeruginosa (Bacillus pyocyaneus), Cya- 

nococcus chroma spirans 

Heinz HUleman, Ber. 7 IB 46 (1938). (Structure) 

G. Farber, Sbornik Ceskoslov. Akad. Zemedelske 23 355 

(1951); Chem. Abstr. 45 9605 (1951). 



Pfizer Handbook of Microbial Metabolites 506 

1001 Cinnabarin (Polystictin), C14H10O5N2, red needles, m.p. : dec. 
>320°. 



HOCH 




Coriolus sanguineus Fr. [= Polyporus cinnabarinus 
Ft. = P. sanguineus Fr. = P. coccineus Fr. = P. puniceus 
Kalch. = Poly stictus cinnabarinus (Jacq.) = P. sanguin- 
eus L. = P. semisanguineus Lloyd = Trametes cinnabarina 
(Jacq.)Fr.] 

About 100 mg. of red pigment were obtained from 55 g. 
of mycelium. 

Jarl Gripenberg, Acta Chem. Scand. 5 590 (1951). 

G. W. K. Cavill, B. J. Ralph, J. R. Tetaz and R. W. Werner, 
J. Chem. Soc, 525 (1953). 

Jarl Gripenberg, Acta Chem. Scand. 12 603 (1958). (Struc- 
ture) 

The same phenoxazone chromophore which occurs in 
cinnabarin and the actinomycins has been found in cer- 
tain insect pigments called ommatins, e.g. xanthommatin : 

HOOC— CH— CH, ^^ ^^^^ 

I I HO COOH 

NH.. C=0 

I 



N 



\ 
O 

Adolf Butenandt, Ulrich Schledt, Ernst Bickert and R. Jan. 
T. Cromartie, Ann. 590 75 (1954). 

1002 Pigment A, C14H11O2N3 -21120, red crystals, dec. without melting. 
Tentative structure: 

CH3 

\e 0OH NH2- HCI 
.N^ /^ / V —COOH 



Yield 12-20 mg. per liter 
and 



507 Phenazines and Phenoxazones 

'003 Pigment B, C,r,H,-,0,iN:iS (may also be hydrated), red crystals, 
dec. without melting. 

An acidic pigment similar to A in structure, but with 
an additional methyl group and a sulfo group. Yield 
30-40 mg. per liter. 

Both produced by a red strain of Pseudomonas aerugi- 
nosa. 

F. G. Holliman, Chem. and Ind., 1668 (1957). 

1004 Griseoiutein A, Ci7Hi40,5No, reddish yellow needles, m.p. 193° 

(dec). 




Streptomyces griseoluteus 

Shoshiro Nakamura, Chem. and Pharm. Bull. (Japan) 6 
547 (1958). 

1005 Griseoiutein B, Ci^HieOgNo, pale yellow crystals, darkening from 
150°, dec. above 220°. Griseoiutein B is a phenazine with 
the following proposed structure : 

COOH OCH3 

ocxp 

CH.— O— CH— CH2 
OH OH 

Streptomyces griseoluteus n. sp. 

Hamao Umezawa, Selki Hayano, Kenji Maeda, Yasuo Ogata 
and Yoshiro Okami, J. Antibiotics (Japan) 4 34 (1951). 

Teisuke Osato, Kenji Maeda and Hamao Umezawa, ibid. 
7A 15 (1954). 

Shoshiro Nakamura, Kenji Maeda, Teisuke Osato and Ha- 
mao Umezawa, ibid. lOA 265 (1957). 

Shoshiro Nakamura, Chem. and Pharm.. Bull. (Japan) 6 
547 (1958). 



Pfizer Handbook of Microbial Metabolites 508 

m. PYRIMIDINES 

Pyrimidines are fundamental components of living 
cells. They have long been recognized as constituents of 
nucleic acids, and more recently other functions have 
been discovered. 

Microorganisms are rather rich in nucleoproteins. 
Yeast, which has been a common experimental source, 
contains about 4 percent of its dry weight in nucleic 
acids, and bacteria up to 15 percent. Bacteriophages are 
largely nucleoprotein, and certain plant viruses entirely. 
By contrast, thymus gland, one of the richer animal tissue 
sources, contains only about 3 percent. 

The protein moieties often are relatively low in molecu- 
lar weight, some of them qualifying as large peptides, and 
they generally seem to be rich in basic amino acids. The 
total nucleoprotein molecular weights, however, are very 
high — often running to many millions. The complexity 
of the nucleic acid moiety varies wdth the complexity of 
the species. Since the DNA carries the genetic informa- 
tion, it might be expected to be more complex and higher 
in molecular weight for the human species than, for ex- 
ample, in a simple plant virus. 

Two types of nucleic acids have been distinguished, 
both widely distributed. Ribose nucleic acid (RNA) and 
deoxyribose nucleic acid (DNA) have been mentioned 
earlier in connection with their roles in protein synthesis 
and genetics. 

Neither of these substances has been obtained entirely 
pure, but newer techniques such as electrophoresis and 
paper chromatography have permitted refinements. The 
important purine and pyrimidine components of RNA are 
adenine, "guanine, cytosine and uracil. In DNA thymine 
takes the place of uracil, and 5-methylcytosine is a minor 



NH 



V Hn"T\ N^l HN 




H HoN H O H O H 

Adenine Guanine Cytosine Uracil 

(6-amino- (2-amino-6- (2-oxy-6-amino- (2,6-dioxy- 

purine) oxypurine) pyrimidine) pyrimidine) 

component in some species. 



509 



Pyrimidines 



CH3 



NH2 



CH3 



NH, 



CH2OH 



) H 



Ai 



Thymine 

(2,6-dioxy-5- 

methylpyrimidine) 



O H 

5-Methyl 

cytosine 

(2-oxy-5-methyl- 

6-aminopyrimidine) 



N II N 

An 

O H 

5-Hydroxymethyl 

cytosine 

(2-oxy-5-hydroxy- 

methyl-6-amino- 

pyrimidine) 

In some Escherichia coli bacteriophages the 5-methyl- 
cytosine is replaced by 5-hydroxymethylcytosine. A sub- 
stance believed to be 5-ribosyluracil has been isolated in 
considerable quantities from yeast RNA. 

The united pyrimidine and ribose moieties are called 
nucleosides, and the phosphorylated nucleosides are called 
nucleotides. 



NH2 





O 

T 

HOCH2 /^\ HO— P— O— CH2 / 

I ^ 

OH 

OH OH OH 

Cytidine Cytidyiic Acid 

(a nucleoside) (a nucleotide) 

The nucleic acids are, then, polymeric nucleotides, a free 
phosphoric acid function being esterified by a free pen- 
tose alcohol group. 

In neither RNA nor DNA are the four main heterocy- 
chc components present in equimolar quantities, and, 
moreover, there is of course species variation. For ex- 
ample, yeast DNA contains more adenine and thymine 
than guanosine and cytosine, while the reverse is true 
for some bacteria. The molar sum of the purines gener- 
ally equals that of the pyrimidines, and, more specifically, 
the number of moles of adenine present equals the num- 
ber of moles of thymine, and the cytosine (and methyl- 
cytosine) equals the guanine. 

There is good evidence now that most DNA is composed 



Pfizer Handbook of Microbial Metabolites 510 

of a helical coil of paired strands, the strands and coils 
being associated by hydrogen bonding, e.g., between the 
amino group of adenine and the carbonyl group of thy- 
mine.^ This structure is supported by roentgen ray dif- 
fraction data, by acid-base titration studies and by light- 
scattering measurements on solutions. There are some 
recent indications, however, that single-stranded DNA 
does exist in some cases. 

Tobacco mosaic virus, a crystalline substance which 
has been investigated extensively, consists of a single 
strand of RNA coiled within a protein sheath. The de- 
gree of organization (non-covalent bonding) in the nu- 
cleic acid moieties of nucleoproteins has been studied. - 
In some instances the nucleic acids seem to be less organ- 
ized in the intact protein than in the free state. 

Pyrimidine nucleotides also serve as coenzymes in a 
number of biological reactions. Thus uridine nucleotide 
is important in the enzymic manipulation of sugars. In 
recent years, uridine-5'-diphosphate sugar esters have 
been isolated from a variety of animal, plant and mi- 
crobial sources. 

Confining our attention to microorganisms, uridine di- 
phosphate glucose, UDP-galactose, UDP-acetylglucosa- 
mine as well as uridine triphosphate (UTP) and uridine- 
diphosphate (UDP) have been isolated from yeast. ^' * •'' 
The same substances have been isolated from Peiiicillium 
chrysogenum mycelium." Other free nucleotides identi- 
fied from the mold were: diphosphophyridine nucleotide 
(DPN), cytidine-5'-monophosphate (CMP), adenosine-5'- 
monophosphate (AMP), triphosphopyridine nucleotide 
(TPN), guanosine-5'-monophosphate (CMP), inosine-5'- 
monophosphate (IMP), uridine-5'-monophosphate (UMP), 

1 J. D. Watson and F. H. C. Crick, Nature 171 737, 964 (1953). 

- F. Bonhoeffer and H. K. Schachman, Biochem. and Biophys. Res. 
Comms. 2 366 (1960). 

• R. Caputto, Luis F. Leloir, C. E. Cardini and A. C. Paladinl, 
J. Biol. Chem. 184 333 (1950); E. Cabib, Luis F. Leloir and C. E. 
Cardini, ibid. 203 1055 (1953). 

■' S. H. Lipton, S. A. Morell, Alexander Frleden and Robert M. Bock, 
;. Am. Chem. Soc. 75 5449 (1953). 

•^Hanns Schmitz, Biochem. Z. 325 555 (1954). 

'• A. Ballio, C. Casinovi and G. Serlupi-Crescenzi, Biochim. et 
Biophys. Acta 20 414 (1956). 



511 Pyrimidines 

adenosine-5'-diphosphate (ADP), guanosine-5'-diphos- 
phatc mannose (GDPM), adenosine-5'-triphosphate (ATP) 
and guanosine-5'-triphosphate (GTP). 

The UTP is an intermediate in the formation of the 
diphosphate: '• "" 

UTP + Sugar-l-phosphate ;=i UDP-Sugar + Pyrophosphate 

Once in the form of UDP esters, sugars are susceptible 
to a variety of enzymic transformations, some of which 
were mentioned in the section on polypeptides. For ex- 
ample, 4-epimerization may be caused : '• '■' 

UDP-Glucose :;=± UDP-Galactose 

and 
UDP-D-Xylose ;=± UDP-L-Arabinose 

Since there is a DPN requirement in these reactions, it 
is likely that the 4-hydroxyl group of the sugar is oxidized 
to a ketone, then reduced stereospecifically. Isotope work 
supports this hypothesis.^"' ^^' ^- UDPG can be oxidized 
also to UDP-glucuronate : '^' ^* 

2DPN 3H^ 

UDP-GIucose — ^ { >UDP-Glucuronic Acid 

H.O 2DPNH 

A yeast enzyme catalyzes the reaction : ^^- ^^ 

UDP-GIucose + Glucose-6-phosphate > Trehalose Phosphate + UDP 

Similarly, di- and polysaccharides seem to be formed 

' Paul E. Trucco, Arch. Biochem. and Biophys. 34 482 (1951). 

^ Agnete Munch-Petersen, Herman M. Kalckar, Enrico Cutolo and 
Evelyn E. B. Smith, Nature 172 1036 (1953). 

•'Luis F. Leloir, Arch. Biochem. aiid Biophys. 33 186 (1951). 

'" Arthur Kowalsky and Daniel E. Koshland, Biochim. et Biophys. 
Acta 22 575 (1956). 

" Laurens Anderson, Aurora M. Landel and Donald F. Diedrich, 
ibid. 22 573 (1956). 

1- Herman M. Kalckar and Elizabeth S. Maxwell, ibid. 22 589 
(1956). 

^■■' V. Ginsburg, E. F. Neufeld and W. Z. Hassid, Proc. Nat. Acad. 
Sci. U. S. 42 333 (1956); V. Ginsburg, /. Biol. Chem. 232 55 (1958). 

^* Evelyn E. B. Smith, Agnete Munch-Petersen and George T. Mills, 
Nature 172 1038 (1953). 

^^E. Cabib and Luis F. Leloir, J. Biol. Chem. 231 259 (1958). 

'•^Luis F. Leloir and E. Cabib, /. Am. Chem. Soc. 75 5445 (1953). 



Pfizer Handbook of Microbial Metabolites 512 

in this way. Involvement in chitin (Neurospora crassay 
and cellulose (Acetobacter xylinumy^ biosynthesis has 
been shown with labeled UDP-acetylglucosamine and 
UDP-glucose, respectively, and work with tritium-labeled 
substrates and cell-free extracts of group A streptococci 
has shown involvement in hyaluronate biosynthesis.^^ 
Other evidence indicates involvement in glucuronide-°' ^^ 
and glycogen--' -^ formation in animals, and glucoside-*' ^^ 
formation in plants. UMP,-« UDP, UTP-'' -^ and UDP- 
glucose-^' •" have been synthesized chemically. 

Several cytidine nucleotides have been isolated from nat- 
ural sources. •■'^' ^-' ^^ CDP-Choline and CDP-ethanolamine 
have been isolated from animals, ^^ plants and yeasts^* 

^' Luis Glaser and David H. Brown, Biochim. et Biophys. Acta 23 
449 (1957); idem., J. Biol. Chem. 228 729 (1957). 

^® Luis Glaser, Biochim. et Biophys. Acta 25 436 (1957); idem., J. 
Biol. Chem. 232 627 (1958). 

^" Alvin Markovitz, J. A. Cifonelli and Albert Dorfman, Biochim. 
et Biophys. Acta 28 453 (1958). 

-" Evelyn E. B. Smith and George T. Mills, Biochim. et Biophys. 
Acta 13 386 (1954). 

21 G. J. Button and I. D. E. Storey, Biochem. J. 57 275 (1954); 59 
279 (1955). 

" Luis F. Leloir and C. E. Cardini, /. Am. Chem. Soc. 79 6340 
(1957); L. F. Leloir, J. M. Olavarria, Sara H. Goldemberg and H. Car- 
minatti, Arch. Biochem. and Biophys. 81 508 (1959). 

"•^ P. W. Bobbins, B. B. Traut and F. Lipmann, Proc. Nat. Acad. 
Sci. U. S. 45 6 (1959). 

-* G. Jacobelli, M. J. Tabone and D. Tabone, Bull. soc. chim.. biol. 
40 955 (1958). 

-> C. E. Cardini and L. F. Leloir, Nature 182 1446 (1958). 

-•^Alexander B. Todd, "Methods in Enzymology" (S. P. Colowick 
and N. O. Kaplan, Editors) Academic Press, New York, 1957 3 p. 811. 

-' B. B. Hurlbert, ibid., p. 785. 

2« G. W. Kenner, A. B. Todd and F. J. Weymouth, /. Chem. Soc, 
3675 (1952); N. Annand, V. M. Clark, B. H. Hall and A. B. Todd, 
ibid., 3665 (1952). 

29 G. W. Kenner, A. B. Todd and B. F. Webb, ibid., 2843 (1954). 

■'" Bobert Warner Chambers, J. G. MofFatt and H. G. Khorana, 
7. Am. Chem. Soc. 79 4240 (1957); J. G. Moffatt and H. G. Khorana, 
ibid. 80 3756 (1958). 

^^ Bolf Bergquist and Adam Deutsch, Acta Chem. Scand. 7 1307 
(1953). 

■" Hanns Schmitz, Bobert B. Hurlbert and Van B. Potter, /. Biol. 
Chem. 209 41 (1954). 

•'•' Eugene P. Kennedy and Samuel B. Weiss, J. Am. Chem. Soc. 77 
250 (1955); idem., J. Biol. Chem. 222 193 (1956). 

^* Irving Lieberman, L. Berger and W. Theodore Gimenez, Science 
124 81 (1956). 



513 Pyrimidrnes 

and seem to be nearly ubiquitous, although so far they 
have not been reported from other microorganisms. 
CDP-Glycerol and CDP-ribitol have been isolated only 
from lactobacilU,''' but probably such substances will be 
found elsewhere. 

CDP-Choline and CDP-ethanolamine are coenzymes es- 
sential to the biosynthesis of lecithin and phosphotidyl- 
ethanolamine.'^ The stages in the biosynthesis of lecithin 
may be outlined : 



2R— CO— S— CoA 




HOCH2CH2N®(CH3)3 
ATP^ 

r^ADP 

POCH2CH2N©(CH3)3 
Cyt— P— P— p«->jr 

ppHI 

Cyt— P— P— OCH2CH2N ® (CHsls 



Cytidine— P + CH2— OCOR 

I 
RCOO— CH O 

I T 

CH2O— P— O— CH2CH2N ® (CHsla 



The cytidine monophosphate can then be rephosphoryl- 

35 J. Baddiley and A. P. Mathias, /. Chem. Soc, 2723 (1954); 
J. Baddiley, J. G. Buchanan, B. Cares, A. P. Mathias and A. R. San- 
derson, Biochem. J. 64 599 (1956). 



Pfizer Handbook of Microbial Metabolites 514 

ated to the triphosphate by ATP, making the process a 
catalytic one. 

The function of the CDP-ribitol and CDP-glycerol in 
Lactobacillus arabinosus seems to be to donate these two 
reduced sugar phosphates in the formation of polymers. 
These ribitol-glycerol-phosphate polymers are components 
of the cell walls of bacteria. Several references are given 
in Appendix A to structural studies on these substances. 

Biosynthesis of the pyrimidines seems to take a similar 
course in microorganisms and in higher animals. So 
many workers have contributed to our knowledge of this 
scheme that referencing cannot be included, but in out- 
line what is now believed to be the important pathway is 
shown below: 

Carbamyl Phosphate 
O 

II 
H2N— C— OPO3H2 H3PO4 

HOOC— CH2— CH— COOH -^^ ^- — ► 

I aspartic transcarbamylase 

NHo 

L-Aspartic Acid 

O 

II 
HO— C 

\ H2O 

H2N CH2 ^ » 

I I dihydro- 

C CH orotase 

O JJ COOH 

N-Carbamyl-l 

aspartic Acid 

(Ureidosuccinic Acid) 



O 



'^\ DPN® DPNH + H® 



HN CHo 



C CH dihydroorotic 

^ \.Ki/^ \ dehydrogenase 



O 'J COOH 

n 

L-Dihydroorotic 
Acid 



515 



Pyrimidities 



5-PRPP PR 



HN 

/^^N^ \ orotidylic 

O H COOH pyrophosphorylase 
Orotic Acid 




COOH 



OH OH 

Orotidine -5 -phosphate 



P— P— P— OCH2 /O 



lOI 



OH OH 

Uridine-5'- 
triphosphote 



CO, 



HN 



n 



'K^J 



ATP 



orotidylic 
decarboxylase POCH.. ,0 




OH OH 
Uridine-5'- phosphate 



HN^l] 



L-glutamine L-glutamate 



AMP 



ATP ADP + P 



NH3 H,0 

cytidylic 
deaminase 



P— P— P— O— CHo ^O 
CMP < 




OH OH 

Cytidine-5'- 
triphosphate 



The biosynthesis of the deoxyribonucleotides may pro- 
ceed similarly as far as uridine-5'-phosphate. Direct 
transfer into the deoxyribose series (i.e. removal of the 
2'-hydroxyl from the ribose moiety) can then occur, or 
hydrolysis to the pyrimidine base and subsequent reaction 
with 2-deoxyribose-l -phosphate can take place. 

There has been much interest in the origin of the 
5-methyl group in thymine (5-methyluracil). The oc- 
currence of 5-hydroxymethylcytosine in some species sug- 
gested donation (in that series) by a tetrahydrofolic acid 
derivative. Isotope experiments indicate that the a-C- 



Pfizer Handbook of Microbial Metabolites 516 

atom of glycine, the /3-C-atom of serine and the C-atom 
of formate can all serve as donors at least indirectly.^*'' ^^ 
There is a vitamin Bio requirement for the conversion of 
formate to the thymine methyl group in Lactobacillus 
leichmannii, and the pathway does not involve methionine 
or a hydroxymethyl group. ^^ It has been suggested that 
since vitamin Bj2 coenzymes are required to promote the 
equilibrium 

HOOC— CH2— CH2— CH— COOH ^ HOOC— CH— CH— COOH 

I 1 I 

NHo CH3 NH2 

Glutamic Acid j3-Methylaspartic Acid 

/3-methylaspartic acid may replace aspartic acid as an 
intermediate in thymine biosynthesis.^'' 

An alternate pathway of pyrimidine biosynthesis in- 
volving dihydrouracil, a member of the catabolic route, 
has been suggested. *° 

O O 

It II /"^ 

An/* /-h\ 

OH O H NH2 

4,5-Dihydrouracil 4,5-Diaminouracil 

The entire subject of the enzymic synthesis of pyrim- 
idines has been reviewed.^^ 

4,5-Diaminouracil has been detected as a metabolite of 
Eremothecium ashbyii and suggested as an intermediate 
in riboflavin biosynthesis. ^- 

■^'^ David Elwyn and David B. Sprinson, 7. Biol. Chem. 207 467 
(1954); idem., J. Am. Chem. Soc. 72 3317 (1950). 

•"J. R. Totter, Elliott Volkin and C. E. Carter, J. Am. Chem. Soc. 
73 1521 (1951); J. R. Totter and Audrey N. Best, Arch. Biochem. 
and Biophys. 54 318 (1955). 

■^^ James S. Dinning, Barbara K. Allen, Ruth Young and Paul L. 
Day, J. Biol. Chem. 233 674 (1958). 

■^^ H. D. Isenberg, E. Seifter and J. I. Berkman, Biochim.. et Biophys. 
Acta 39 187 (1960). 

^" Lewis C. Mokrasch and Santiago Grisolia, Biochim,. et Biophys. 
Acta 27 227 (1958). 

•" Peter Reichard, Advances in Enzymology 21 263-294 (1959). 

*-T. W. Goodwin and D. H. Treble, Biochem. J. 67 lOp (1957). 



5 1 y Pyrimidines 

1006 Uracil, C4H4O2N2, colorless needles, m.p. ^-'335° (dec.)- 

O 

II 

O H 

Agaricus nehularis, yeasts 

Nils Lofgren, Bjorn Liining and Harry Hedstrom, Acta 
Chem. Scand. 8 670 (1954). 

1007 Cytosine, C4H5ON3, large colorless crystals, m.p. ^320° (dec). 

NH2 



N 

O H 

Agaricus nehularis 

Nils Lofgren, Bjorn Liining and Harry Hedstrom, Acta 
Chem. Scand. 8 670 (1954). 

1008 4,5-Diaminouracil, C4H6O2N4, has been shown to be a metabolite 
of Eremothecium ashbyii by trapping with diacetyl. 



HN 



O' 

O H 



NH, 



NH2 



T. W. Goodwin and D. H. Treble, Biochem. J. 67 lOp (1957). 

1009 Uridine, CgHjoOgNs, colorless crystals, m.p. 165°, [a]D^° +6.4° 
(10°) (in water). 



HOCH2 



"O 

/^N-^ 



O 




OH OH 



Pfizer Handbook of Microbial Metabolites 



518 



Yeast 

Hellmut Bredereck, Annelise Martini and Friedrich Richter, 
Ann. 74 694 (1941). 

Hubert S. Loring and James McT. Ploeser, /. Biol. Chem. 
178 439 (1949). 

1010 Cytidine, C9Hi30riN3, colorless needles, m.p. 225-230° (dec), 
[alo'" +29.6° (in water). 



HOCH 




OH OH 



Yeast 

HeUmut Bredereck, Annelise Martini and Friedrich Richter, 
Ann. 74 694 (1941). 

1011 Uridine-3'-phosphate (Uridylic Acid), C^Hj^OoNoP, colorless 
prisms, m.p. 200° (dec), [a]„ +9.5 to 14.5° (in water). 



HN 



HOCH 
O 

(OHl^P O 




OH 



Yeast 

The 5'-di- and triphosphates also have been isolated 
from microorganisms. 

Hellmut Bredereck and Gerd Richter, Ber. 7 IB 718 (1938). 

W. E. Cohn and C. E. Carter, J. Am. Chem. Soc. 72 2606 
(1950). 

A. M. Michelson and A. R. Todd, /. Chem. Soc, 2476 (1949). 



519 



Pyrimidines 



1012 C:vtidine-2'-phosphate (Cytidylic Acid) CuHi^OhN-,?, colorless 
crystals, m.p. 238-240° (dec), [a]u +20.7° (c 1.0 In 
water). 




Yeast 

Hubert S. Loring, Nydia G. Luthy, Henry W. Bortner and 
Luis W. Levy, /. Am. Chern. Soc. 72 2811 (1950). 

Hubert S. Loring and Nydia G. Luthy, ibid. 73 4215 (1951). 

1013 Cytidine-3'-phosphate (Cytidylic Acid), C,,Hi40sN;{P, colorless 
tablets, m.p. 230-234° (dec), [ali, +49°. (c 0.5 in water). 




(OHIjP O OH 



Yeast 

The 5'-di- and triphosphates also have been isolated 
from microorganisms. 

Hubert S. Loring, Nydia G. Luthy, Henry W. Bortner and 
Luis W. Levy, /. Am. Chem. Soc. 72 2811 (1950). 

Hubert S. Loring and Nydia G. Luthy, ibid. 73 4215 (1951). 



Pfizer Handbook of Microbial Metabolites 



520 



1014 Orotidine (Orotic Acid Riboside), CioHioOgNg, cyclohexylamine 
salt, m.p. 183°. 



O 
HOCH2 ^o 



COOH 




OH OH 



Neurospora crassa mutant 

A. Michael Michelson, William Drell and Herschel K. Mitch- 
eU, Proc. Nat. Acad. Sci. U. S. 37 396 (1951). 



1015 Cytidine Diphosphate Glycerol, C12H21O12N3P2. 



NH2 



CH2— CH— CHo 

I 1 

OH OH 




OH OH 



Lactobacillus arabinosus 

J. Baddiley and R. P. Mathias, /. Chem. Soc, 2723 (1954). 
J. Baddiley, J. G. Buchanan, B. Cares, A. P. Mathias and 
A. R. Sanderson, Biochem. J. 64 599 (1956). 

1016 Cytidine-5'-diphosphatecholine ( CDP-Choline ) , C13H04O11N4P2, 
amorphous white, hygroscopic powder. 



CH3 CH3 



O O ^^ 

\l II II An 

N— CH2— CH2— O— P— O— P— O— CH2 O 

/e II 

CH3 O® OH 



OH OH 



521 



Pyrimidines 



Yeast 

This compound is a biogenetic precursor of the lecithins 
and cephahns. 

Irving Lieberman, L. Berger and W. Theodore Gimenez, 
Science 121 81 (1956). 

Eugene P. Kennedy and Samuel B. Weiss, J. Biol. Cheni. 
222 193 (1956). 

1017 Cytidine Diphosphate Ribitol, Cj^Ho-PigNaPs. 



O O N 

T T /^N 

CH2— CH— CH— CH— CH2— O— P— O— P— O— CHo O 



OH OH OH OH 



OH 



O 
H 



OH OH 



Lactobacillus arabinosus 

J. Baddiley and A. P. Mathias, /. Chem. Soc, 2723 (1954). 
J. Baddiley, J. G. Buchanan, B. Cares, A. P. Mathias and 
A. R. Sanderson, Biochem. J. 64 599 (1956). 



1018 Uridinediphosphateglucose (UDPG), C15H04O17N2P2. 

OH 
O 



CH2OH 

A — o. 



K \l o o 

koH yi II II 

HO N K O— P— O— P- 



-0— CHo 



OH 



OH OH 




OH OH 



Yeast, molds 

A uridinediphosphateacetylglucosamine also has been 
isolated from yeast. 

R. Caputto, Luis F. Leloir, C. E. Cardini and A. C. Paladini, 
J. Biol. Chem. 184 333 (1950). 

E. Cabib, Luis F. Leloir and C. E. Cardini, ibid. 203 1055 
(1953). 

J. G. MofFatt and H. G. Khorana, /. Am. Chem. Soc. 80 3756 
(1958). (Synthesis) 



Pfizer Handbook of Microbial Metabolites 



522 



1019 Thymidine Diphosphate Rhamnose, C16H26O14N2P2. 



OH 



HO i— O. 
/CH3 \ O 






OH OH 



O— P— O— P— O— CH2 /^^N 




OH OH 




CH3 



Lactobacillus acidophilus 

Reiji Okazaki, Biochem. and Biophys. Res. Comms. 1 34 
(1959). 

1020 Plicacetin (Amicetin B), C2r,H3-,07N5, colorless needles, m.p. 
182-184° from H.O— CH3OH, [aW +181° (c 2.7 in 
methanol). 



CH3 CH3 



OH 



HOCHo 



CH3 



^7- 



/- 



NH2 



Streptomyces plicatus 

Theodore H. Haskell, Albert Ryder, Roger P. Frohardt, Sal- 
vatore A. Fusari, Zbigniew L. Jakubowski and Quentin R. 
Bartz, ;. Am. Chem. Soc. 80 743 (1958). 



523 



Pyrimidines 



1021 Bamicetin, CosH^oO,,N,., white microcrystals, m.p. 240° (dec), 
[a],,-"" +123° (c 0.5 in 0.1 N hydrochloric acid). 
Partial Structure: 



C13HJ4O5N 



\/N 



■r 

N 



NH-C-/ \ 
O 



O CH3 
NH— C— C— CH2OH 
NH2 



Streptomyces plicatus 

Theodore H. Haskell, Albert Ryder, Roger P. Frohardt, Sal- 
vatore A. Fusari, Zbigniew L. Jakubowsjci and Quentin R. 
Bartz, ;. Am. Chem. Soc. 80 743 (1958). 

1022 Amicetin (Sacromycin, Allomycin), C29H42O9N6, colorless nee- 
dles, m.p. 165-169°, [a],.-' +116.5° "(c 0.5 in 0.1 N hy- 
drochloric acid). 



Amosamine 



CH, 



[ HOCH> 



CH3 




OH 



CH3 



CH3 



0^\ /=N 
N // 



/ 



O 

NH— C- 



O CH3 

II I 
NH— C— C— CH2OH 

NH, 



a-Methyl- 
D-serine 



Cytosine 



p-Aminobenzoic 
Acid 



Streptoviyces vinaceus-drappus, S. fasciculatus, S. sin- 
denensis, S. plicatus 

Edwin H. Flynn, J. W. Hinnan, E. L. Caron and D. O. Woolf, 
Jr., ;. Am. Chem. Soc. 75 5867 (1953). 



Pfizer Handbook of Microbial Metabolites 524 

Calvin L. Stevens, Robert J. Gasser, Tapan K. Mukherjee 
and Theodore H. HaskeU, ibid. 78 6212 (1956). 



n. PURINES 

The nature of nucleic acids and the participation of 
purines in their structure were discussed in the preceding 
section. The process of oxidative phosphorylation also 
was mentioned although it is not yet entirely understood. 
In this process inorganic phosphate ions disappear dur- 
ing biological oxidation of substrates and become bound 
in adenosine triphosphate (ATP), the universal storage 
molecule for chemical energy wdthin cells. Many ex- 
amples of ATP as an energy donor were seen in earlier 
sections. 

Adenosine polyphosphates have other functions, most 
of them concerned with the activation and transfer of 
various chemical moieties with formation of new chemi- 
cal bonds. ATP, for example, can donate phosphate or 
pyrophosphate groups to form new phosphate esters. 
Two such known reactions are: 



hexokinase 
Glucose + ATP ;===^ Glucose-6-phosphate + ADP 

M^ 
and phosphoribose 

pyrophosphokinase 
Ribose-5-phosphate -H ATP ^ ^ri Ribose-5-phosphate-l- 

pyrophosphate + AMP 



Adenosine-3'-phospho-5'-phosphosulfate has been estab- 
Ushed as activated sulfate,^- - and it has been used in the 
formation of sulfate esters of a number of phenols and 

^ Robert S. Bandurski, Lloyd G. Wilson and Craig L. Squires, 
/. Am. Chem. Soc. 78 6408 (1956). 

2 P. W. Robbins and Fritz Lipmann, ibid. 78 2652, 6409 (1956). 



525 



Purines 





NH2 

1 




o 

r 

HO— S- 

i 

o 




T 

0— P— 0— CH 
OH 


^ 




Adenosine-3'-phospho- 
5'-phosphosulfate 




1 1 
OH 

1 






HO 


— P— OH 

i 








alcohols in the presence of sulfokinases. The generality 
of the sulfate transfer mechanism has been demonstrated 
in yeast, neurospora and liver. 

The recognition of S-adenosylmethionine as the active 
complex in methyl group transfer from methionine (and 
perhaps in its biosynthesis) was noted in the section on 
amino acids. 

In the section on aliphatic acids an ATP requirement 
was noted in the formation of acyl coenzyme A. A num- 
ber of acyl adenylates have been prepared or isolated 
from natural sources.^' "*■ ^ These can be converted en- 
zymically into acyl coenzyme As. The general structure 
of these activated acids is: 



R— C- 



NH2 



O 

T 

-0— P— O— CH; 

I 

OH 



&^^ 



K^ 



OH OH 



Acid Anhydrides of 
Adenosine-5'-pliosphate 



3 Paul Berg, ibid. 77 3163 (1955). 

■* Preston T. Talbert and F. M. Huennekens, ibid. 78 4671 (1956). 

•■^C. H. Lee Peng, Biochim. et Biophys. Acta 22 42 (1956). 



Pfizer Handbook of Microbial Metabolites 



526 



In the same section the mediation of ATP in the forma- 
tion of active carbon dioxide was seen: 



NH2 



ATP + CO2 

O O 

II T 

HO— C— O— P— O— CH 
OH 



S.Xn'^ 



+ Pyrophosphate 



OH OH 



I Biotin phosphate 



NH2 



O 

II 



o 00 

II T T 

C— O— O— P— O— P— O— CH> .0 






HN 

1 
CH- 



N 

I 
-CH 



OH OH 




OH OH 



CH. CH2— (CH,)4— COOH 

\c/ i 

Enzyme 



Possible intermediate in 

formation of activated 

carbon dioxide 



Synthetic adenosyl-5'-phosphoryl carbonate has been pre- 
pared.*^ 

The role of adenine nucleotide as the terminal or ac- 
tivating nucleotide of transfer RNA in protein synthesis 
was mentioned in the amino acid section. 



*^ B. K. Bachhawat, J. F. Woessner and M. J. Coon, Federation 
Proc. 15 214 (1956). 



527 Purines 

Finally, the occurrence of the adenine nucleotide moiety 
in various other coenzymes (coenzyme A, flavine-adenine 
dinuclcotide, DPN, etc.) should not be forgotten. The 
functions of these coenzymes are considered elsewhere. 

Adenine polyphosphates, then, are so ubiquitous and so 
metabolically important that they nearly all have been 
encountered prior to this point in our discussions of mi- 
crobial metabolism. 

Guanosine polyphosphates, too, are widespread, and 
they seem to be able to duplicate some of the less specific 
functions of those of adenine. One reaction in which a 
guanine polyphosphate is known to participate is : ' 



a-Ketoglutaric Acid + DPN© + CoA-SH -^ 

Succinyl-S-CoA + DPNH + H© + CO2 
Succinyl-S-CoA + Guanosine Diphosphate + H3PO4 ;=^ 

Succinic Acid + CoA-SH + GTP 



The enzyme catalyzing this reaction has been isolated 
only from tissues of higher animals, and there is evidence 
that in Escherichia colt at least the adenine nucleotide 
seems to be involved.'' 

Guanosine and inosine nucleotides also participate in 
the formation of phosphoenolpyruvate from oxaloace- 
tate : " 

Oxaloacetic Acid + GTP :;=± Phosphoenolpyruvic Acid + GDP + COn 

but again this has been shown only in animal tissues. 

The general function of GTP as an energy source in 
the amination of inosinic acid during adenine biosynthe- 
sis will be seen later. 

Guanosine diphosphate mannose has been isolated 

" D. R. Sanadl, David M. Gibson, Padmaslni Ayengar and Miriam 
Jacob, ;. Biol. Chem. 218 505 (1956). 

^ Roberts A. Smith, Irma F. Frank and I. C. Gunsalus, Federation 
Proc. 16 251 (1957). 

»M. F. Utter and K. Kurahashi, /. Biol. Chem. 207 821 (1954). 



Pfizer Handbook of Microbial Metabolites 



528 



from yeast^° and a penicillium mold^^ as well as from 
higher animals, and it probably occurs in plants. Guano- 



OH 




O O H2N 

T T 
I— O— P— O— P— O— CH2 ^o. 



OH OH 



OH OH 







OH OH 



Guanosine Diphosphate Mannose 



sine diphosphate fucose has been isolated from Aerobacter 
aerogenes,^'- and this organism has an enzyme which con- 
verts GDP-mannose to GDP-fucose. This conversion re- 
quires TPNH and must involve several steps to accom- 
plish the requisite epimerizations and reduction of the 
terminal carbon atom. The functions of these guanosine 
derivatives are unknown, but yeast elaborates a mannan, 
and fucose is a proven constituent of bacterial polysac- 
charides (as well as blood group specific polysaccharides 
in higher animals). This may then be a form in which 
sugars are modified and transported for incorporation 
into polysaccharides. 

A substance of the vitamin B12 group isolated from 
Nocardia rugosa has been identified as guanosine diphos- 
phate factor B, i.e. a guanosine-5'-pyrophosphoric ester of 
factor B in which ribose is linked to N-9 of guanine (par- 
tial structure shown ).^^ 

10 E. Cabib and Luis F. Leloir, ibid. 206 779 (1954). 

^^ A. Ballio, C. Casinovi and G. Serlupi-Crescenzi, Biochim. et 
Biophys. Acta 20 414 (1956). 

12 V. Ginsburg and H. N. Klrkman, J. Am. Chem. Soc. 80 3481, 
4426 (1958). 

'^^ R. Barchielli, G. Boretti, A. DlMarco, P. Julita, A. Migliacci, 
A. Minghetti and C. Spalla, Biochem. J. 74 382 (1960). 



529 



Purines 



CN 

>f< 

CN 

° r "'"^ 

0<-P— O— P— O— CH . 

I I 

OH OH 



OH 




Guanosine Diphosphate 

Factor B 

(Factor B = Vitamin B12 

minus the 

dimethylbenzimidazole 

nucleotide moiety) 



OH OH 



This substance has been suggested as an intermediate 
near the end of the vitamin B12 synthesis just prior to 
introduction of the dimethylbenzimidazole nucleotide. 

There is evidence that labeled guanine is an isotopic 
precursor of riboflavin in Eremothecium ashbyii. Ade- 
nine also is a precursor of this vitamin. In each case 
the Cs atom is lost. In the case of adenine, at least, the 
pyrimidine ring is incorporated intact into riboflavin^* 
although pyrimidines such as uracil and thymine are in- 
effective precursors." 

Inosine is an intermediate in the biosynthesis of ade- 
nine and guanine, but beyond the phosphoenol pyruvate 
formation and some of the less specific reactions of the 
purine nucleotides (phosphate transfer, etc.) few func- 
tions have been discovered. 

The purine nucleotides have been reviewed.^" ^^' ^*- ^^- ^°- ^^ 

i-* Walter S. McNutt, Jr., /. Biol. Chem. 219 365 (1956). 

15 John A. MacLaren, /. Bacteriol. 63 233 (1952). 

1'' Paul D. Boyer, Henry Lardy and Karl Myrback, "The Enzymes" 
Vol. II, Robert M. Bock, Adenine nucleotides and properties of pyro- 
phosphate compounds. Academic Press, New York, 1960, pp. 3-27. 

1" Ibid., Merton F. Utter, Guanosine and inosine nucleotides, pp. 
75-87. 

18 Jack L. Strominger, Physiol. Rev. 40 55-111 (1960). 

"J. Baddiley and J. G. Buchanan, Quart. Rev. 12 152-172 (1958). 

-° Standish C. Hartman and John M. Buchanan, Advances in En- 
zymology 21 199-261 (1959). (Copyright 1959 by Interscience 
Publishers, Inc., New York) 

21 G. E. W. Wolstenholme and Cecilia M. O'Connor (Eds.), "CIBA 
Foundation Symposium on the Chemistry and Biology of Purines," 
J. M. Buchanan, J. G. Flaks, L. C. Hartman, B. Levenberg, L. N. 
Lukens and L. Warren, The enzymatic synthesis of inosinic acid 
de novo. Little, Brown and Co., Boston, 1957, pp. 233-255. 



Pfizer Handbook of Microbial Metabolites 



530 



The general scheme of purine biosynthesis is under- 
stood now. It is outlined in the following equations :" 



O3POCH2 




OH OH 

Ribose-5- 
phosphate 



ATP AMP O3POCH2 o 
OH Mg^H) 




OPocS 



Glut- Glut- 
amine amate 

V ^ 



Mg<S> 



OH OH 
Ribose- 1 -pyrophos- 
phate-5-phosphate © 

NH3 

/ 
CH2 



a 



03POCH2 




Glycine ADR 

+ +©fi 

NH. ATP HPO4 O3POCH2 



c=o 

I 
NH 



OH OH 
1-Aminoribose- 
5'-phosphate 



OH OH 

Glycinamide 
Ribotide 



H2O 



CH2 CHO 



N\Ni°-Anhydro- 
- formyl THFA 

■— ^THFA 
.NK 



© I 
H2N=C 



a 




Glut- Glut- 
note amii 
k Mg(H>; 



CH. 

c=o 



CHO 



'O3POCH0 r. NH amate amine ""O3POCH2 ^ NH 




OH OH ADP ATP 

Formylglycin- HPO4© H;0 

amidine Ribotide 



OH OH 

Formylglycin- 
amide Ribotide 



H2O ^1 ^^ ATP 
HP04Q^^^-^ADP 



" Reproduced from reference 20. 



531 



Purines 







i 
HC- 



V 



CH 



©. 



H,N' 



OaPOCHj 




OOC 



XD3POCH, 



\^. 



CH 



CO. 



biotin 




OH OH 
Aminoimidazole 
Ribotide 



OH OH 

5-Amino-4-imid- 

azolecarboxylic 

Acid Ribotide 



O 

II 

,c. 



HoN 



^, 



CH 



© H.N' 

O3POCH2 




Fumarate 



OH OH 
5-Amino-4-imidazole- 
carboxamide Ribotide 



Aspartate--^ I ^ ATP + H2O 

S..ADP + HPOr 



COO® 

I 

CH2 

I 

CH— NH- 
COO® 



o 
-c 



:. H2N' 

OaPOCH. 



/ 




OH OH 

5-Amino-4-imidazole- 

N-succinocarboxamide 

Ribotide 



Sulfanilamide and other sulfa drugs inhibit the growth 
of many bacteria by interfering with the incorporation 
of p-aminobenzoic acid into the folic acid coenzymes 
(p-aminosalicylic acid, etc., may do the same in mycobac- 
teria), and E. coli cultures so inhibited accumulate isola- 
ble quantities of 5-amino-4-imidazolecarboxamide ribo- 
tide.-' 

2^ Joseph S. Gots and Edith G. GoUub, Proc. Nat. Acad. Sci. U. S. 
43 826 (1957). 



Pfizer Handbook of Microbial Metabolites 532 

Azaserine, a glutamine antagonist, inhibits purine syn- 
thesis in some bacteria, and causes accumulation of for- 
mylglycinamide ribotide in E. coli.-* Another antibiotic, 
6-diazo-5-oxo-L-norleucine, also inhibits purine biosynthe- 
sis at this stage. Purine-requiring mutants of E. coli and 
A. aerogenes accumulate the following compounds or 
derivatives : aminoimidazole,-'^ 5-aminoimidazolecarboxa- 
mide,-'' 5-amino-4-imidazole-N-succinocarboxamide-^ and 
xanthine.-' Yeast grown on a biotin-deficient medium 
gives off aminoimidazole riboside and hypoxanthine.-^ 

Cell-free extracts of Neiirospora crassa are able to pro- 
mote all the reactions shown in the biosynthetic scheme 
above. All these facts as well as other evidence indicate 
that this is the principal biosynthetic route to purines in 
bacteria and fungi, and probably is quite general. 

Inosinic acid is an intermediate in the biosynthetic 
route to the other purines as shown in the formula se- 
quence on page 533. 

Extracts of Aerobacter aerogenes convert inosinic acid 
to xanthylic acid, and there is other evidence that the 
final stages of purine biosynthesis follow this route in 
many bacteria and fungi as well as in animal cells. 

Other references can be found in some of the reviews 
of this subject.-"' ^^ 

There are indications that methylated purines may be 
minor constituents of yeast and bacterial nucleic acids. 
Traces of 6-methylaminopurine, 6-hydroxy-2-methylami- 
nopurine and 1-methylguanine were detected in yeast 
RNA.-^ Small amounts of 6-methylaminopurine, 6,6-di- 

-^A. J. Tomisek, H. J. Kelley and H. E. Skipper, Abstr., 128th 
Meeting, Am. Chem. Soc, 5C, Minneapolis, Sept., 1955. 

25 Samuel H. Love and Joseph S. Gots, J. Biol. Chem. 212 647 
(1955). 

26 Joseph S. Gots, ibid. 228 57 (1957). 

2^ Boris Magasanik, H. S. Moyed and Lois B. Gehring, ibid. 226 
339 (1957). 

2* D. P. Lones, C. Rainbow and J. D. Woodward, /. Gen. Microbiol. 
19 146 (1958). 

2^^ Max Adler, Bernard Weissmann and Alexander B. Gutman, 
;. Biol. Chem. 230 717 (1958). 



533 



Purines 



e © 

OOC— CH2— CH— coo 

I 

NH, 



HN 






©O3POCH2 






Aspartate GDP 

+ GTP +HP04^ 



OH OH 
Inosinic Acid 



MgO 



•^3POCH2 



DPN® 

+ HjO ^ 

DPNH 
+H© 



:® 



)>L^ 



OH OH ^^ 

Adenylosuccinic Fumarote 

Acid 

NH2 



©O3POCH2 



*/0 



OH OH 
Adenylic Acid 



HN 



O H 



©O3POCH 



OH OH 



OH 



Xanthylic 
Acid 



Glutamine, Glutamate, 
ATP, H2O AMP, HP2O7© 



NHj, ATP AMP, HP2O7S 




N' ir % 



H2N 



©O3POCH2 



OH OH 

Guanylic 
Acid 



Pfizer Handbook of Microbial Metabolites 



534 



methylaminopurine and 2-methyladenine have been 
found in bacterial RNA.^° 



CH3 
NH 

H H 

6-Methylamino- 6-Dimethylamino- 
purine purine 




OH 




CH;,— NH H 

2-Methylamino-6- 
hydroxy purine 




NH, 



CH,— NH 



^N^^N- 
H2N H 

1-Methylguanine 




-N^^N^ 
CH3' H 

2-Methyladenine 




Kinetin is a substance isolated from yeast which stim- 
ulates cell division in plant tissues. Work on kinetin and 
related compounds has been reviewed. ^^ 

Several antibiotics contain the purine nucleus. Some 
of these have excited interest as purine analogues for 
tumor inhibition, but they are all toxic. Puromycin is 
an inhibitor of protein synthesis. - The interference has 
been shown to occur at the last stage — that is the ex- 
change of the activated amino acid between transfer- 
RNA and the growing protein chain. 

3f'J. W. Littlefield and D. B. Dunn, Biochem. J. 68 8P (1958); 
idem.. Nature 181 254 (1958). 

2^ E. R. Squibb Lectures on Chemistry of Microbial Products, "Top- 
ics in Microbial Chemistry," John Wiley and Sons, New York, 1958, 
F. M. Strong, Kinetin and kinins, pp. 98-158. 

"-Michael Yarmolinsky and Gabriel de la Haba, Chem. and Eng. 
News April 25, 1960. 



535 



Purines 



CH3 CH 



HOCH 




NH2 




HOCH2 




NH OH 




OH OH 


C— CH— CH,- 


_// y 


-OCH3 


NH2 






Puromycin 




Adenine 
Nucleoside 



Substitution of other amino acids for the p-methoxy- 
phenylalanine moiety gives analogues which still inhibit 
protein synthesis, although the free nucleoside moiety 
is a less effective inhibitor. The similarity in structure 
suggests competition with adenine nucleoside. 

Functions of coenzyme A have been discussed through- 
out the appropriate sections. The biosyntheses of the 
various moieties of the molecule also have been consid- 
ered with the possible exception of ^-aminoethanethiol, 
which is derived from cysteine. 

The biosynthetic union of these moieties, originally 
studied in animal tissues, follows the probable course: 



CH3 OH 

HOCH2— C CH— COOH 

CH3 
Pantoic Acid 



+ HoN— CHo— CH2— COOH 

^ATP 
— >ADP 



^-Alanine 



Pfizer Handbook of Microbial Metabolites 



536 



Cysteine 




i 
CH3 OH O 

I I II 
HOCH2— C CH— C— NH— CHo— CH2— COOH 

CH3 

Pantothenic Acid 



O 

T 



COOH 
Pantothenylcysteine 

CH3 OH O 

HOCH2— C CH— C— NH 

I I 

CH3 CHz 

I 
CHz 

I 
HSCH2— CH2NHC=0 

Pantetheine 

ATP 
ADP 





CH3 OH O 

HO— P— O— CH2— C CH— C- 

I I 

OH CHs 



Pantothenic Acid 
4'-Phosphate 

r'^Cysteine 

CH3 OH O 

I I II 
HO— P— O— CH2— C CH— C 

I I 

OH CH3 



NH 

I 
CHz 

CH2 

COOH 



o 

T 



NH 

I 
CH2 

I 
CH2 

I 
HSCHz- CH— NH— C=0 

COOH 

Pantethenylcysteine- 
4'-Phosphate 




O CH3 OH O ^ O 

T I I II 

HO— P— O— CH2— C CH— C— NH— CH2— CH2— C— NH— CH2— CH2SH 

OH CH3 

Pantetheine-4'-phosphate 

11 ATP 



537 



Purines 




11 

o o 

T T 

CHo— O— P— O— P— O— CH2 
OH OH 



CH3 OH O 

I I II 
-C CH— C— NH 

CH3 I 

CH2 

I 
CH2 

HSCH2— CH2— NH— C=0 
3'-Dephosphocoenzyme A 

i ATP 

Coenzyme A 

Most of these intermediates have been identified in mi- 
croorganisms, e.g. Streptobacterium plantar urn. '-^^ Pan- 
tothenic acid is required by some microorganisms, but 
probably not by man, perhaps because of the excess pro- 
duced by E. coli and other intestinal microbes. 

A number of higher fungi and molds have been ex- 
amined thoroughly for nucleotide content. Some of the 
organisms which have been studied are: Penicillium 
chrysogenum,^* Aspergillus oryzae/'' Polyporus squamo- 
sus,^^ Amanita muscaria,^^ Lycoperdon pratense,^^ 
Hypholoma capnoides/'^ Armillaria mellea,^^ Pholiota 
squarrosa,^'^ Lactarius vellereus,^''' Lactarius turpis,^'^ Toru- 
lopsis utilis,^'' Micrococcus lysodeikticus,^^ Coprinus co- 
matis,^^ and Polyporus sulfureus.*^ 

^3 Theodor Wieland, Walter Maul and Ernst Friedrich Moller, 
Biochem. Z. 327 85 (1955). 

^* A. Ballio, C. Casinovi and G. Serlupi-Crescenzi, Biochim. et 
Biophys. Acta 20 414 (1956); Alessandro Ballio and Giovanni Serlupi- 
Crescenzi, Nature 179 154 (1957). 

^^ Kazuo Okunuki, Kozo Iwasa, Fumlo Imamoto and Tadoyoshi 
Higashiyama, J. Biochem. (Tokyo) 45 795 (1958). 

3«Rolf Bergkvlst, Acta Chem. Scand. 12 1549, 1554 (1958). 

37 D. Gilbert and E. Yemm, Nature 182 1745 (1958). 

38 J. V. Scaletti, Dissertation Abstr. 17 1191 (1957). 

39 Paul Heinz List, Arch. Pharm. 291 502 (1958). 
*°ldem., Planta Med. 6 424 (1958). 



Pfizer Handbook of Microbial Metabolites 538 

1023 Hypoxanthine, C5H4ON4. 



H 

Amanita muscaria. Boletus edulis, Agaricus nebularis, 
Polyporiis sulfiireus 

E. Buschmann, Pharm. Post 45 453 (1912). (Chem. Abstr. 
6 2485) 

E. Winterstein, C. Reuter and R. Korolev, 7. Chem. Soc. 
104 I 433 (1913). 

Nils Lofgren, Bjorn Liining and Harry Hedstrom, Acta 
Chem. Scand. 8 670 (1954). 

Paul Heinz List, Planta Med. 6 424 (1958). 

1024 Xanthine, C5H4O0N4, colorless crystals, m.p. 220° (dec). 

O 






H H 

Amanita muscaria 

E. Buschmann, Pharm. Post 45 453 (1912). (Chem. Abstr. 
6 2485 ) 

1025 Uric Acid, C-,H403N4, colorless crystals, m.p. >400° (dec). 

O 

OH 

Aspergillus oryzae 

Miazuko Sumi, Biochem. Z. 195 161 (1928). 

1026 Adenine, C-.H-.N., (Trihydrate), colorless needles, m.p. 360-365° 

(dec) (subl. from 220°) (Picrate), dec. 280°. 




539 Purines 

Coprinus comatis Gray, Boletus edulis, Polyporus sul- 
fureus 

Paul Heinz List, Arch. Pharrn. 291 502 (1958). 

E. Winterstein and C. Reuter, Centr. Bakt. Parasitenk. II 
Abt. 34 566 ( 1912). {Chem. Abstr. 6 3279) 

Paul Heinz List, Planta Med. 6 424 (1958). 

1027 Guanine, C3H5ON5, (Picrate) dec. from 190°. 




HN 



Coprimis comatis Gray, Boletus edulis 
Paul Heinz List, Arch. Pharin. 291 502 (1958). 
E. Winterstein, C. Reuter and R. Korolev, /. Chem. Soc. 104 
I 433 (1913). 

1028 Heteroxanthine, CgHeOoN^, colorless crystals, m.p. ~380° 

(dec). 

O ^^^ 

O H 

Yeast 

P. W. Wiardi and B. C. P. Jansen, Rec. trav. chim. 53 205 
(1934). 

1029 Toxoflavin, C6H6O2N4, yellow crystals, m.p. 171°. 



CH3 

N 

An 

O H 

Pseudomonas cocovenenans 

A. G. van Veen and W. K. Mertens, Proc. Acad. Sci. Amster- 
dam 36 666 (1933). (Isolation) (Chem. Abstr. 27 5771) 

A. G. van Veen and J. K. Baars, Rec. trav. chim. 57 248 
(1938). (Structure) 



Pfizer Handbook of Microbial Metabolites 



540 



1030 



Kinetin ( 6-Furf urylaminopurine ) , C10H9ON5, colorless prisms, 
m.p. 265° (sealed tube to prevent sublimation). 




Yeast extracts 

E. R. Squibb Lectures on Chemistry of Microbial Products, 
"Topics in Microbial Chemistry," John Wiley and Sons, New 
York, 1958, F. M. Strong, Kinetin and kinins, pp. 98-157. 

1031 Nebularine (9-(/3-D-Ribofuranosyl) purine), C10H12O4N4, color- 
less prisms, m.p. 181°, [alo^^ —48.6° (c 1 in water). 



N 



HOCH2 



k:^ 



OH OH 

Agaricus (Clitocybe) nebularis Batsch. 

Lars Ehrenburg, Harry Hedstrom, Nils Lofgren and Bertil 
Takman, Svensk Kem. Tidskr. 58 269 (1946). 

Nils Lofgren, Bjorn Liining and Harry Hedstrom, Acta 
Chem. Scand. 8 670 (1954). 

David L Magrath and George Bosworth Brown, J. Am. 
Chem. Soc. 79 3252 (1957). (Synthesis) 

1032 Cordycepin, C10H13O3N5, colorless needles, m.p. 225°, [<xW° 
—47° (in water). 




HOCH2 



541 Purines 

Cordyceps militaris (Linn.) Link 

K. G. Cunningham, S. A. Hutchinson, William Manson and 
F. S. Spring, J. Chem. Soc, 2299 (1951). 

H. R. Bentley, K. G. Cunningham and F. S. Spring, ibid., 
2301 (1951). (Structure) 

1033 Adenosine, CioHi^O^N^, needles, m.p. 229°, [aW -60 to -63° 
(in water). 




HOCH2 ^ 



OH OH 



Agaricus nehularis 

Nils Lofgren, Bjorn Liining and Harry Hedstrom, Acta 
Chem. Scand. 8 670 (1954). 

1034 Guanosine, C10H13O5N5, colorless crystals, m.p. 237° (dec), 
[alD'" -60° (in 0.1 N sodium hydroxide). 




H:N 



HOCH, Q 



OH OH 



Yeast 

Hellmut Bredereck, Annelise Martini and Friedrich Richter, 
Ber. 74B 694 (1941). 



Pfizer Handbook of Microbial Metabolites 



542 



1035 Inosine-5'-phosphate (Inosinic Acid), C10H13O8N4P, a syrup. 

OH 




O 

II 
(OHliP— OCH2 




OH OH 



Yeast, Penicillium chrysogenum 

The 5'-diphosphate also has been isolated. 

E. Cabib, Luis F. Leloir and C. E. Cardini, /. Biol. Chem. 
203 1055 (1953). 

A. Ballio, C. Casinovi and G. Serlupi-Crescenzi, Biochim. et 
Biophijs. Acta 20 414 (1956). 

1036 Adenosine-2'-phosphate (Adenylic Acid a), C10H14O7N5P, color- 
less crystals, m.p. 187° (dec). 



NH" 






HOCH2 



^i 



HO 



O— P(OH)2 

II 

o 



Yeast 

D. M. Brown, G. D. Fasman, D. I. Magrath, A. R. Todd, 
W. Cochran and M. M. Woolfson, Nature 172 1184 (1953). 

C. E. Carter, J. Am. Chem. Soc. 72 1466 (1950). 

Joseph X. Khym, David G. Doherty, Elliot Volkin and 
Waldo E. Cohn, ibid. 75 1262 (1953). 

D. M. Brown and A. R. Todd, /. Chem. Soc, 44 (1952). 



543 



Purines 



1037 Adenosine-3'-phosphate (3-Adenylic Acid, Yeast Adenylic Acid), 
C„,H,40;N-,P, colorless crystals, m.p. 191-195° (dec), 
[a]i.-" —66° (c 2 in 5', sodium hydroxide). 



NH2 




Yeast, Penicillium chrysogenum 

H. Steudel and E. Peiser, Z. physiol. Chem. 127 262 (1923). 
D. A. Kita and W. H. Peterson, J. Biol. Chem. 203 861 
(1953). 

1038 Adenosine-5'-phosphate (Muscle Adenylic Acid), Ci(jHi407N5P, 
colorless crystals, m.p. 178°, [aju"" —50° (in formamide). 



NH.. 



OH 

0=P— O— CH. 

I 
OH 




OH OH 

Yeasts, Lactobacillus arabinosiis, Penicillium chryso- 
genum 

The 5'-diphosphate (ADP) also has been isolated from 
microorganisms. 

E. Cabib, Luis F. Leloir and C. E. Cardini, ]. Biol. Chem. 
203 1055 (1953). 

J. Baddiley and A. C. Mathias, /. Chem. Sac, 2723 (1954). 

A. Ballio, C. Casinovi and G. Serlupi-Crescenzi, Biochim. et 
Biophys. Acta 20 414 (1956). 



Pfizer Handbook of Microbial Metabolites 



544 



1039 Guanosine-3'-phosphate (Guanylic Acid), C10H14O8N5P, colorless 
crystals, [ajo -7.5° to -13.5° (in water). 



H.N 






(OH)2P— O 
O 



OH 



Yeast 

The 5'-di- and triphosphates also have been isolated 
from microorganisms. 

Walter Jones and M. E. Perkins, /. Biol. Chem. 62 557 
(1925). 

1040 Adenosine-5'-triphosphate (ATP), CioHigOia N5P3. 




OH OH OH 

I I I 

0=P— O— P— O— P— OCH > 

I il II " 

OH O O 




OH OH 



Yeasts, molds, bacteria, etc. (widely distributed) 

Th. Wagner-Jauregg, Z. physiol. Chem. 238 129 (1936). 
(Isolation) 

G. A. LePage and W. W. Umbreit, J. Biol. Chem. 148 255 
(1943). 

D. A. Kita and W. H. Peterson, ibid. 203 861 (1953). 

A. Endo, Ann. Report Takamine Lab. 11 45 (1959). 



545 



Purines 



1041 Angustmvcin A, CjiH,304N.r,, colorless needles, m.p. (anhydr.) 

169.5° (dec.), [<xW^ +48.3°. 
Probable structure: 






— C 

II 
C— OH 

I 
H— C— OH 

I 
H— C— OH 

1 

- C— H 

I 
CH3 



Adenine 



>L-2-Ketofucopyranose 



Streptomyces hygroscopicus 

Hsii Yiintsen and Hiroshi Yonehara, Bull. Agr. Chem. Soc. 
(Japan) 21 261 (1957). 

Hsii Yiintsen, Kazuhiko Ohkuma, Yoshio Ishil and Hiroshi 
Yonehara, J. Antibiotics (Japan) 9A 195 (1956). (Isolation 
and characterization) 

Hsii Yiintsen, ibid. IIA 79 (1958). (Structure) 

1042 Angustmycin C (Psicofuranine), C11H15O5N5, colorless crystals, 
m.p. 202-204°, [aW^ -71.1° (c 1.8 in pyridine). 



NH2 



N \f 



HOCHo 




CH2OH 



OH OH 



Streptomyces hygroscopicus var. angustmyceticus 
Hsu Yiintsen, /. Antibiotics (Japan) llA 244 (1958). 
(Structure) 



Pfizer Handbook of Microbial Metabolites 



546 



1043 Nucleocidin, CnH^eOgNfjS, colorless crystals, no definite m.p., 
[a],r*'' -33.3° (c 1.05 in 1 : 1 ethanol, 0.1 N hydrochloric 
acid). 
Partial structure: 






NHo 

I 
CeHioOs 

I 
OSOoNH, 



CeHioOs is an unusual 
reducing sugar. 



Streptomyces calvus 

S. O. Thomas, V. L. Singleton, J. A. Lowery, R. W. Sharpe, 
L. M. Pruess, J. N. Porter, J. H. Mowat and N. Bohonos, "An- 
tibiotics Annual 1956-1957," Medical Encyclopedia Inc., New 
York, p. 716. (Isolation) 

C. W. Waller, J. B. Patrick, W. Fulmor and W. E. Meyer, 
J. Am. Chem. Soc. 79 1011 (1957). (Structure) 

1044 Adenylosuccinic Acid, Ci4HisOiiN.r5P, no properties listed. 



HOOC— CH— CH.— COOH 



NH 



H2O3POCH. 



OH OH 



Penicillium. chrysogenum (mycelium) 

About 16 known derivatives of adenine, guanine, cyti- 
dine, uracil, etc., also were detected in this study. 

Alessandro Ballio and Giovanni Serlupi-Crescenzi, Nature 
179 154 (1957). 



547 



Purkies 



1045 Diadenosinctetraphosphate, C2„H^.„0i.,Ni„P, [a]r,44o"" —39.2° (in 
N sulfuric acid). 



NH; 



OH OH OH 

I I I 

0=P— O— P— O— P— O— CH 

I II II 

OH O O 



-I '2 



OH 



-O— P— O— CH 



OH 




OH OH 



Yeast 

W. Kiessling and O. Meyerhof, Natunvissenschaften 26 13 
(1938). 

1046 Coenzyme A, C^iHaeOjcN-SPa, white amorphous powder. 



NHo 








CH2— O- 


CH3 OH 
-P— 0— P— 0— CH2-C CH— C— NH 


OH C 


> 
\ 


P(OH); 

i 



OH OH CHj CH2 

1 
CH2 

1 
HSCHo— CH2— NH— C=0 



Occurs widely in microorganisms and higher animals. 
Yeast and certain streptomycetes were early sources. 

F. M. Strong, "Squibb Lectures on the Chemistry of Mi- 
crobial Products," Coenzyme A and related compounds, John 
Wiley and Sons, Inc., New York, 1956, pp. 44-98. (This re- 
view lists 117 earlier references.) 



Pfizer Handbook of Microbial Metabolites 



548 



J. G. Moffatt and H. G. Khorana, /. Am. Chem. Soc. 81 1265 

(1959). (Synthesis) 

1047 Puromycin, C22H29O5N7, white crystals, m.p. 175.5-177° 
(uncorr.), [a]D^^ —11° (c 1 in ethanol). 



HOCH 




0==C— CH— CH 



OCH3 



Streptomyces albo-niger 

J. W. Porter, R. I. Hewitt, C. W. Hesseltine, G. Krupka, J. A. 
Lowery, W. S. Wallace, N. Bohonos and J. H. Williams, Anti- 
biotics and Chemotherapy 2 409 (1952). 

Coy W. Waller, Peter W. Fryth, Brian L. Hutchings and 
James H. Williams, /. Am. Chem. Soc. 75 2025 (1953). 
( Structure ) 

B. R. Baker, Robert E. Schaub, Joseph P. Joseph and 
James H. Williams, ibid. 77 12 (1955). (Synthesis) 



0. PTERIDINES AND FLAVINES 

Pteridines (pterins), originally discovered in insects, 
occur widely, and several have been isolated from mi- 
crobial sources. The most important of these from the 
metabolic standpoint is folic acid. This substance, or 
group of related substances, is a vitamin for most mam- 
mals and plants and for some microorganisms unable to 
produce it. Pure folic acid first was isolated from liver 
and from yeast. The triglutamyl form was isolated from 
a corynebacterium, and the heptaglutamyl derivative, first 
isolated from yeast, since has been found in a variety of 
microorganisms. The reason for the polypeptide chains is 
not clear. These forms are as effective as folic acid in 



549 



Pteridines and Flavines 



higher animals, but are not so active as folic acid for the 
bacteria ordinarily used in bioassays. 

The functions of folic acid as a B-vitamin have been in- 
vestigated extensively and are now largely understood. 
Some of these have been encountered earlier in our discus- 
sions, but the role of folic acid derivatives in one-carbon 
metabolism has not been considered as such. 

In its coenzyme form folic acid is attached to a protein 
apoenzyme, probably at the glutamic acid moiety, and the 
pteridine ring is reduced. One of these pteroproteins has 
been crystallized.^ The "active formate" form of the co- 
enzyme has been shown to be N^'--formyltetrahydrofolic 
acid,^- ^' ^ and the "active formaldehyde" form probably is 
N'',N^'^-methylenetetrahydrofolic acid.^' ^- ''• * 



HoN 




/^N^-^N^ 



"Active Formaldehyde" N^ N^"- 
Methylenetetrahydrofolic Acid 

OH 



\Xj 



CHO 
CH.— N ^^^— C— I 



HoN 



O COOH 

I 
NH— CH 

1 
CH2 

I 
CH2 



COOH 



"Active Formate" 
N^^-Formyltetrahydrofoiic Acid 



^ Jesse C. Rabinowitz and W. E. Pricer, Jr., Federation Proc. 17 
293 (1958). 

- H. M. Rauen and Lothar Jaenicke, Z. physiol. Chem. 293 46 
(1953). 

3 Lothar Jaenicke, Biochim. et Biophys. Acta 17 588 (1955). 

4H. M. Rauen, Biochem. Z. 328 562 (1957). 

5R. L. Blakley, Biochem. J. 58 448 (1954). 

«Roy L. KisHuk, J. Biol. Chem. 227 805 (1957). 

^ M. J. Osborn and F. M. Huennekens, Biochim.. et Biophys. Acta 
26 646 (1957). 

^ F. M. Huennekens and M. J. Osborn, Advances in Enzym.ology 21 
370 (1959). 



Pfizer Handbook of Microbial Metabolites 550 

The two forms are interconvertible and this oxidation- 
reduction equilibrium probably is mediated by an enzyme 
with triphosphopyridine nucleotide (TPN) as the pros- 
thetic group. 

Formate added as a substrate is, then, activated in this 
way. The N^"-formyl group also can be furnished by 
glycine, either by way of glyoxylic acid'^ ^^ or by way of 
S-aminolevulinic acid.^^ ^'' ^^ The equations are: 



transam- 
ination [O] 

H2N— CH2— COOH , OHC— COOH * 

Glycine Glyoxylic 

Acid 

COo -f Ni°-Formyltetrahydrofolic Acid 
and 

o 00 

li NH3 II II 

H,N— CH,— C— CH,— CH2— COOH . ^ ' H— C— C— CH,.— CHo— COOH 
6-Aminolevulinic Acid a-Ketoglutoraldeiiyde 

[O] 
^ HOOC— CH>— CH2— COOH + N'o-Formyltetrahydrofolic Acid 

Succinic Acid 



Once formed "active formate" is the formylating agent 
in certain metabolic reactions. The important formyla- 
tions by this agent which have been discovered to date are 
the two formylations already noted in the biosynthetic 
route to the purines. Thus glycineamide ribotide is 
formylated to furnish C-8 of the purine nucleus and, later, 
5-amino-4-imidazolecarboxamide ribotide is formylated to 
furnish C-2 of the purine nucleus. 

^ Henry I. Nakada and Sidney Weinhouse, Arch. Biochem. and 
Biophijs. 42 257 (1953). 

1^' Sidney Weinhouse in W. D. McElroy and H. B. Glass (Editors), 
"Amino Acid Metabolism," Johns Hopkins Press, Baltimore, 1955, pp. 
637-57. 

" David Shemin, ibid., p. 727. 

^^ David Shemin, Tessa Abramsky and Charlotte S. Russell, /. Am. 
Chem. Soc. 76 1204 (1954). 

'"^ Irving Weliky and David Shemin, Federation Proc. 16 268 
(1957). 



551 



Pteridines and Flavines 



CH.NHj 

\ 
NH 



NH 
/ \ 
CH, CHO 

O C 

\ 
NH 



P— O— CH, 




"Active Formate" 



P— 0~CH, 




OH OH 

Glycineamide 
Ribotide 



and 



OH OH 

Formylglycineamide 

Ribotide 



O 

II 

c 



HoN 



-N 

II 
CH 



H2N 



P— O— CH, 




"Active 
Formate" 

K® 



H,N 
OHC 

P-O— CH, 



-N 
CH 



HN 



OH OH 

5-Amino-4-imidazole- 
carboxamide Ribotide 



OH OH 

5-Formamido-4-imidazole- 
carboxamide Ribotide 



As was seen in the biosynthesis of histidine the N-1 and 
C-2 atoms of the purine nucleus are donated to this amino 
acid during its formation so that 



NH2 

N— 1 I 

N Y 



C— 2 



^^N^N 



N N 

1 T 
c— 2 


-CH- 
NHo 


-COOH 







Adenine 



Histidine 



indirectly, at least, these atoms too are furnished by the 
coenzyme. 



Pfizer Handbook of Microbial Metabolites 



552 



The "active formaldehyde" form of the coenzyme is in- 
termediate in the interconversion of glycine and serine: 

"Active 
Formaldehyde" 



CH2— COOH ^ 



± HOCH2— CH— COOH 



NH2 



NH2 



Glycine 



The large literature on this subject has been reviewed.^ 

The "active formaldehyde" form may also be considered 
to be a methyl group donor, although much remains to be 
learned about the mechanisms of these donations. In the 
biosynthesis of thymine from uracil, serine, formaldehyde 
or formate are more effective precursors of the introduced 
methyl group than is methionine, and this precursor effect 
is inhibited by foUc acid antagonists.® Actually, the ac- 
ceptor is probably not uracil, but deoxyuridine or deoxy- 
uridylic acid: 




OH 

2'-Deoxyuridine 
5'-phosphate 



OH 

2'-Deoxy-5-methyiol- 
uridine-5'-phosphate 



Thymidine- 
5'-phosphate 



The occurrence of 5-hydroxymethylcytosine in some spe- 
cies has been cited as suggestive of formation of a hy- 
droxymethyl intermediate in this way, at least in the 
cytosine series."- ^^ On the other hand it has been re- 
ported that in Lactobacillus leicJimamiii there is a vitamin 
B12 requirement for the conversion of formic acid to the 
thymine methyl group, and that the route does not involve 
either methionine or a hydroxymethyl group. ^"^ 

" Seymour S. Cohen and Lawrence L. Weed, J. Biol. Chem. 209 
789 (1954). 

15 Maurice Green and Seymour S. Cohen, ibid. 225 387 (1957). 

1*^ James S. Dinning, Barbara K. Allen, Ruth Young and Paul L. 
Day, ibid. 233 674 (1958). 



553 



Pteridines and Flavines 



The synthesis of the labile methyl group of methionine 
has been shown to involve a one-carbon unit at the form- 
aldehyde oxidation level, and the "active formaldehyde" 
form of the coenzyme has been implicated.'"' '■ Here, 
again, not everything is known. The following route has 
been suggested : ''• ^^ 



€' 






HOOC CH— CHo—CH.— S— CH, 



NH2 



OH OH 

S-Adenosylhomocysteine 



"Active 

Formaldehyde" 



Methionine 



Homo- 
cysteine 



e 

00c— CH— CH2— CH2— S— CH. 



NH2 




OH OH 

S-Methylol-S-adenosylhomocysteine 



TPNH + H® 



G 

OOC— CH— CH,— CHo— S— CHs 



NH2 




OH OH 

S-Adenosylmethionine 

1" David Elwyn, Arthur Weissbach and David B. Sprinson, J. Am. 
Chem. Soc. 73 5509 (1951). 

18 David B. Sprinson in W. D. McElroy and H. B. Glass (Editors), 
"Amino Acid Metabolism," Johns Hopkins Press, Baltimore, 1955, p. 
608. 

I'' Audrey Stevens and W. Takami, Federation Proc. 17 316 (1958). 



Pfizer Handbook of Microbial Metabolites 554 

In an Escherichia coli mutant requiring either methi- 
onine or vitamin B12 for growth methionine synthesis 
from homocysteine and serine was stimulated by addition 
of vitamin 612-""' '^ This suggests that again vitamin Bjo 
may be involved in methyl group synthesis. 

There is some evidence (from higher animals) that 
there is a folic acid requirement for the introduction into 
aminoethanol of some, if not all, of the methyl groups of 
chohne.-" '-^^ 

Little is known about the biosynthesis of pteridines in 
microorganisms. There are suggestions that both pteri- 
dines and flavines are related to the purines in this respect. 



•rr^ 



7 4I 5 CH 



Pteridine 



>N T ^6 






COOH 


2— NH— / V-C— NH- 


-CH 




CH2 

1 




1 
CH2 


Pteroyl-L-glutamic Acid 
(Folic Acid) 


COOH 



Labeled molecule studies with butterflies indicate that 
carbon atoms 4 and 5 of the pteridine ring in leucopterin 
and xanthopterin are derived from glycine (4 from the 
glycine carboxyl group and 5 from the a-carbon atom).-^ 

OH ■■ ^ O" 

O I OH 



N j ] 



N 



H2N H O H2N 

Leucopterin Xanthopterin 

The C-6 position seems to be furnished from carbon diox- 
ide and the C-2 position from formate, reminiscent of the 
purines. Carbon atoms 8 and 9 of the pteridine nucleus 

20 c. W. Helliner and D. D. Woods, Biochem. J. 63 26 p (1956). 
-1 R. L. Kisliuk and D. D. Woods, J. Gen. Microbiol. 18 xv (1957). 
^- Jacob A. Stekol, Sidney Weiss and Ethyl I. Anderson, /. Am. 
Chem. Soc. 77 5192 (1955). 

23 R. Venkataraman and D. M. Greenberg, ibid. 80 2025 (1958). 
2* F. Weygand and M. Waldschmidt, Angew. Chem. 67 328 (1955). 



555 



Pteridines and Flavines 



(in leucopterin from butterflies) are furnished quite di- 
rectly by glucose. Over 50 percent of the activity of 
D-glucose-1-C" was found in these two positions, and 
acetate was excluded as a direct precursor of this part of 
the molecule.-^' 

A sugar origin for this part of the pteridine ring is sug- 
gested, too, by the natural occurrence of such substances 
as erythropterin and biopterin, although, in these cases, 

OH OH 
OH _ OH 



N ^ 



H.N 



OH 



C=C— CH> 



CH— CH— CH3 



N 



^^ 



N^^N 



HoN 



OH OH OH 

Erythropterin Biopterin 

pentoses would be expected. Both erythropterin and 
biopterin, incidentally, occur as glycosides. If a precursor 
such as this were assumed, it would relate these sub- 
stances closely with the riboflavin structure. There is 
experimental support for the assumption of the pyrimi- 
dine shown as a riboflavin precursor.^" 




NH. 



CH2 

1 

CH— CH— CH— CH2 

1111 
OH OH OH OH 



O 




CHj 



CH3 



CH2 

I 
CH— CH— CH— CH2 

I I I I 

OH OH OH OH 



Assumed pteridine 
precursor 



Riboflavin 



Many pteridine derivatives related to the pteridine 

-' F. Weygand, H.-J. Schliep, H. Simon and G. Dahms, ibid. 71 522 
(1959). 

-^ Toyokazu Kishi, Mitsuko Asai, Toru Masuda and Satoru Kuwada, 
Chem. and Pharm. Bull. (Japan) 7 515 (1959). 



Pfizer Handbook of Microbial Metabolites 556 

moiety of folic acid have been isolated from non-microbial 
species. This subject has been reviewed.^' '^ 

Labeled xanthopterin was converted to 5-formyl-5,6- 
7,8-tetrahydropteroic acid by Enterococcus stei. Strepto- 
coccus fecalis, E. coli and Pichia membranaefaciensr''' 
Folic acid was not formed even when p-aminobenzoic acid 
was added to the medium. Cell extracts of these micro- 
organisms produced folic acid principally. 

The assembly of the three moieties of folic acid into the 
complete molecule has been studied. Lactobacillus arabi- 
nosus contains enzymes able to couple 2-amino-4-hydroxy- 
pteridine-6-carboxaldehyde or the corresponding alcohol 
with p-aminobenzoic acid.-^ 

OH OH 

I .. .CHO I . /CH2OH 






H2N H2N 

2-Amino-4-hydroxy- 2-Amino-4-hydroxy-6- 

pter'dine-6-carboxaldehyde hydroxy methylpteridine 

These pteridines are even more effective precursors in 
their reduced forms. Many other pteridines tested were 
not used. ATP (and Mg*"^) was required. Its role is un- 
known, although phosphorylation of the alcohol of the 
pteridine hydroxymethyl group might be necessary to 
activate it for coupHng. 

p-Aminobenzoic acid was more effective than p-amino- 
benzoylglutamic acid in this coupling reaction in E. coli,^^ 
although Mycobacterium avium was able to use the pep- 
tide.^" Apparently adenylo-p-aminobenzoic acid was an 
intermediate in the latter organism (ATP and Co A were 
required ) . 

The origin of p-aminobenzoic acid was considered in an 
earlier section. It has been known for some time that the 
anti-infective sulfonamide drugs function by interfering 

27 J. J. PfifFner and O. D. Bird, Ann. Rev. Biochem. 25 416-^19 
(1956). 

27' F. Korte and Gotthard Synnatschke, Ann. 628 153 (1959). 

28 T. Shiota, Arch. Biochem. and Biophys. 80 155 (1959). 
2«Gene M. Brown, Federation Proc. 18 19 (1959). 

^° H. Katunuma, Abstr. 32nd Congr. Japanese Biochem. Assoc, 
Kyoto, July 1957. 



557 



Pteridines and Flayines 



with the incorporation of p-aminobenzoic acid into fohc 
acid. Enzyme studies (E. coli extracts) now seem to have 
narrowed this to inhibition of the couphng of the pteridine 
moiety with p-aminobenzoic acid," although in the Myco- 
bacterium avium study inhibition of peptide formation by 
prevention of adenylo-p-aminobenzoic acid formation was 
suggested. 

Investigation of the biosynthesis of riboflavin is facili- 
tated by the existence of the two microorganisms, Eremo- 
thecium ashbyii, a yeast, and Ashbya gossypii, a mold, 
which are prodigious producers of this vitamin, evolving 
large quantities into the culture medium. 

Besides riboflavin several other substances have been 
isolated from riboflavin fermentations. The structures of 
these metabolites suggest that they may be biosynthetic 
precursors of the vitamin. 



CH3 



\ 



c=o 

I 
CH-OH 



CH3 



HN 




NH2 



H NH., 



^V^ 



CH3 






CHs 



CH2— CH— CH— CH— CH2 



OH OH OH OH 

Acetoin 4,5-Diaminouracil 6,7-Dimethyl-8-(D- 1 '-ribityl)-lumazine 

G-Compound (green fluorescence) 



3HN 





OH OH OH OH 

6-Methyl-7-oxy-8-(D- 1 '-ribitylj-lumazine 
V-Compound (violet fluorescence) 



OH OH OH OH 

Riboflavin 



They are shown in the accompanying formulas. 

Addition of purines to cultures of growing riboflavin 
producers increases the yield of riboflavin.''- C"-8-Labeled 
adenine contributes no radioactivity to the riboflavin mole- 

■'*i Gene M. Brown, Physiol. Revs. 40 359 (1960). 
32 John A. MacLaren, /. Bacteriol. 63 233 (1952). 



Pfizer Handbook of Microbial Metabolites 



558 



cule,"'^ but C-4 of the purine nucleus is equivalent to C-4a 
in riboflavin, and C-5 of purine to C-9a of riboflavin.^"* 
The C-4 of riboflavin is furnished by carbon dioxide (cf. 
C-6 in purines), and C-2 from formate (cf. C-2 in pu- 
rines). These relationships are shown in generalized 
diagram. 



Sources of the Carbon Atoms in Purines, Pteridines and Flavines 



co; 

i 



o 



HCOOH 



y 



IN iT| 



\ 



HCOOH HCOOH 



Purine 




HCOOH 



The pyrimidine rings in all these systems seem to have 
a common origin, and perhaps purines are precursors of 
the other two classes of heterocycles. 

Guanine-5-C^ * was converted to labeled riboflavin and to 
labeled G-compound by Eremothecium ashbyii, Ashbya 
gossypii, Candida fiareri, C. guilliermondii and C. parapsi- 
lopsis.'^-' Pyrimidines and pteridines were not used di- 
rectly, and, when labeled G-compound was added to grow- 
ing cultures, it was not converted to riboflavin by E. 
ashbyii nor was labeled 4,5-diaminouracil. V-Compound 
was shown to be formed rather easily from G-compound 
by air oxidation of a stored alkaline solution. While G- 
compound was not used by growing whole cells, cell-free 
extracts of E. ashbyii, Ashbya gossypii, Mycobacterium 
smegmatis and M. avium were able to incorporate it into 
the riboflavin molecule.-^'" ■" 

33 Walter S. McNutt, 7. Biol. Chem. 210 511 (1954). 

3*G. W. E. Plaut, ibid. 208 513 (1954). 

"•"' Friedhelm Korte, Hans Ulrich Aldag, Gerhard Ludwig, Wilfried 
Paulus and Klaus Storiko, Ann. 619 70 (1958). 

3« Friedhelm Korte and Hans Ulrich Aldag, Ann. 628 144 (1959). 

3" G. F. Maley and G. W. E. Plaut, /. Am. Chem. Soc. 81 2025 
(1959). 



559 



Pteridines and Flavines 



Adenine was found to be a more efficient precursor for 
riboflavin than G-compound in C"-labeling studies,-^"^ and 
guanine and xanthine have been found more efficient than 
adenine. •' 

Acetate'" and shikimic acid" have been shown to be im- 
probable direct precursors of the A ring of riboflavin. 
Acetoin has been isolated from riboflavin fermentations*- 
and is a normal metabolite of these organisms and of 
other yeasts. On the basis of chemical studies this sub- 
stance (or near derivatives) was proposed as a precursor 
of the A ring of riboflavin.''^ '* It has been confirmed that 
acetoin is an efficient biological precursor of the vitamin" 
although intermediates cannot be ruled out entirely. 

At present, then, the following biosynthetic scheme 
seems indicated: 



Purine, 

Purine nucleoside 

or 
Purine nucleotide 




r^' 



HO 




NH 












CH, 




Pyruvic Acid 

i 






(CHOH)3 


"Active Acetaldehyde 






1 




i 






CH2OH 


Acetoin 








\y 










--♦ 


2H2O 













CH3 




Acetoin 


II 


CH3 


""^Y"^^ 




)\o] 


H\f^ 


f^ Y^ 


-N--^-^ 


3H2O 


„>M^ 


^N^ 


1 CH, 






1 CH3 


CHo 








CH2 


(CHOH), 








(CHOH)3 


CH2OH 










CH2OH 



3-^ R. Cresswell and H. Wood, Proc. Chem. Soc, 386 (1959). 

3»E. G. Brown, T. W. Goodwin and S. Pendleton, Biochem. J. 68 
40 (1955). 

40 G. W. E. Plaut, ;. Biol. Chem. 211 111 (1954). 

"T. W. Goodwin and D. H. Treble, Biochem. J. 70 14 p (1958). 

42Toru Masuda, Pharm. Bull. (Japan) 5 136 (1957). 

«A. J. Birch and C. J. Moye, /. Chem. Soc, 412 (1957); 2622 
(1958). 



Pfizer Handbook of Microbial Metabolites 560 

The occurrence of V-compound could be explained as due 
to a side-reaction in which pyruvate rather than acetoin 
reacted with the pyrimidine, or it may merely be an oxida- 
tion product of G-compound. The close relationship be- 
tween pyruvate, active acetaldehyde and acetoin, which is 
mediated by thiamine, has been discussed in an earlier 
section. 

The origin of the ribityl group remains obscure. It is 
yet to be shown whether this moiety is derived from the 
ribose of the purine nucleosides or whether it is formed in 
some other way. Some work has been done on this facet 
of the biosynthesis. ^'^ **• *^' ^'^ 

Riboflavin is phosphorylated by ATP to give riboflavin- 
5'-phosphate, a coenzyme form. This, in turn, can react 
again with ATP in the presence of the appropriate en- 
zyme to form flavine-adenine dinucleotide, the other co- 



Mg++ 

Riboflavin -f ATP -^ Riboflavin-5'-phosphate + ADP 

Riboflavin-5'-phosphate -|- ATP :^ Flavine-adeninedinucleotide + Pyrophosphate 



enzyme form. Flavine-adenine dinucleotide (FAD) is 
produced commercially in Japan from E. ashbyii my- 
celium. 

The principal point of attachment of fiavinemononucle- 
otide (FMN) to the apoenzyme seems to be the phosphate 
group. There may be involvement of the 3-imino group 
also. FAD is the most prevalent coenzyme form, although 
FMN occurs in rather large proportions in some microor- 



44 G. W. E. Plaut and Patricia L. Broberg, /. Biol. Chem. 219 131 
(1956). 

4=^ Edna B. Kearney and Sasha Englard, ibid. 193 821 (1951). 

4^ Anthony W. Schrecker and Arthur Kornberg, ibid. 182 795 
(1950). 



561 Pteridines and Flavines 

ganisms. Obligate anaerobes contain relatively large 
quantities of flavoproteins. Surveys have been made of 
the flavine content of microorganisms not used in com- 
mercial production.'' '^ There is variation in the tight- 
ness of binding of the coenzyme, and the modes of attach- 
ment are not entirely understood. 

One of the functions of the flavine enzymes has been 
mentioned already, namely, the dehydrogenation of re- 
duced DPN in the respiratory chain. Sites of DPNH for- 
mation were seen earlier, particularly in the glycolysis 
route and the citric acid cycle. The enzyme succinic de- 
hydrogenase is a flavoprotein, and the FADH^ formed in 
this reaction also is fed into the respiratory chain. Besides 
the direct net synthesis of 2 moles of ATP during glycoly- 
sis and of 1 mole of ATP in the citric acid cycle, the re- 
maining energy released during glucose catabolism is 
transferred in the form of hydrogen or electrons to en- 
zymes with TPN, DPN or FAD as prosthetic groups. 

These reduced enzymes are, in turn, oxidized by the 
metal ion-porphyrin enzymes, which are oxidized by gase- 
ous oxygen. When two hydrogen atoms are passed along 
the entire respiratory chain, water is formed as well as 3 
more molecules of ATP. 

The exact number of particles in the chain is not en- 
tirely clear, and there are variations with different or- 
ganisms. In lactobacilli, for example, flavines seem to 
replace heme proteins in electron transport. *^^ Also ob- 
scure is the exact manner in which ATP is formed during 
respiration and the precise way in which hydrogen is 
transferred from one coenzyme to the next. There has 
been interesting speculation in this area of biophysics. 

The respiratory chain can be shown in a simplified form 
as in the accompanying diagram. ^^ 



''"J. L. Peel, Biochem. J. 69 403 (1958). 

*^ Chester DeLuca, Morton M. Weber and Nathan O. Kaplan, 
/. Biol. Chem. 223 559 (1956). 

^s" Cornelius F. Strittmatter, Federation Proc. 17 318 (1958). 
■*» Albert L. Lehninger, Scientific American 202 102 (1960). 



Pfizer Handbook of Microbial Metabolites 



562 



s 
u 

B 




/" 



^iSH 



Jx <— dpnh4x 








>'r> 



+ 

H 

The natures of the entities X, Y and Z are mysterious. If 
they are assumed to possess nucleophiUc groups such as 
R — S', R — COO" or H0PO4", then one scheme has been ad- 
vanced to show how the requisite energy-rich bonds could 
be formed in DPNH, FADH and Ferricytochrome ag.^" 
The couphng methods and resonance systems involved are 
shown in the diagram: 



CHj 



H Hilll 
Rb-ADP 



H H -K 



/ +HX;=; r 'J; ^N; + oh© 



^N 

Rb-ADP 



C H 

N +2e -f- HjO 



H % (3. 

I ill! " 



^0(2) 



CHj 1 r ^>ri,„ CH: 

ADP-Rb H 
(1) 



H X 







HC— X 



CjHs CH; 

CH Fe(3) CH + HX 

fatty CH 

acid II 

RCH 





ADP-Rb ^) 

+2e + H2O 



CH— 



-f-OHQ— a 



Z 



OH 



R— C -t- HS— E ;^ R— C— SE -> R— C -f 2e -f- 2H© 

\ I \ 

H H SE 



5" Paul E. Glahn and Sigurd O. Nielsen, Nature 183 1578 (1959). 



563 



Pteridines and Flavincj 



CYT 


2 


CYT 
C" 



H-f'-'U 



&0*-' 



zh^2nr:!Hz 2 



CYT 







+ 
H 

Another hypothesis assumes close approach of DPNH 
and riboflavin in parallel planes with interposition of in- 
organic phosphate, held perhaps by hydrogen bonding, 
e.g. to the amide moiety of nicotinamide.'^^ These geomet- 
rical and chemical relationships can be represented as 
follows : 




ie " 



"Barbro Grabe, Biochim. et Biophys. Acta 30 560 (1958). 



Pfizer Handbook of Microbial Metabolites 564 

When an electron is transferred from the N-atom of the 
reduced pyridine ring to an unoccupied 7r-orbital of the 
isoalloxazine ring of FAD, the N-atom assumes a positive 
charge, which is neutrahzed by attraction of a proximate, 
ionized phosphate hydroxyl oxygen. The increased elec- 
tron density on the 0-atom at position 2 in the riboflavin 
nucleus might cause formation of a bond to phosphorus 
as shown in the activated complex above, the reaction 
being : 

DPNH + FAD + H2O3PO® + H® ^ DPN® + H2O3P— FADH + OH® 

When this substance is oxidized by the subsequent carrier 
(probably a cytochrome), two electrons, perhaps dislocal- 
ized TT-electrons, are withdrawn from the FAD-complex 
thus permitting dissociation of a proton and activation of 
the phosphoryl group. In the presence of ADP, then, ATP 
could be formed according to the equation: 

FADH— PO3H2 + ADP + 2Fe€I5) ^ FAD + H© + ATP + 2Fe<£5 

Other flavoprotein dehydrogenase substrates are: alde- 
hydes, a-amino acids, a-hydroxy acids, purines, fatty acid- 
coenzyme A esters and certain amines. Flavine enzymes 
also participate in bacterial hydrogenase systems, in ni- 
trate reduction and assimilation by fungi and higher 
plants and in photosynthesis and bioluminescence. There 
is currently much study of flavoprotein reactions, which 
can often be followed by spectrophotometry and EPR tech- 
niques. 

Reviews of the flavine coenzymes and their biosynthesis 
are available. ^^' ^^ 

1048 Xanthopterin, CgHjOoNg, yellow amorphous substance, isolated 
as barium or sodium salts. 

OH 

I .OH 



HoN 



N Y 1 



^2 Paul D. Boyer, Henry Lardy and Karl Myrback (Eds.), "The 
Enzymes" Vol. II, 2nd ed., Helmut Beinert, Flavin coenzymes. 
Academic Press, New York, 1960, pp. 340-416. 



565 Pteridines and Flavines 

Mycobacterium tuberculosis 

Also occurs as a butterfly wing pigment. 

Marguerite 0"L. Crowe and Amy Walker, Brit. J. Exptl. 
Pathol. 35 18 (1954). (Isolation from this organism) 

Robert Purrmann, Ann. 546 98 (1940), 548 284 (1941). 
(Synthesis) 

1049 Pterin-like Substance. 

By paper chromatographic comparisons this purple 
fluorescent substance was shown to be similar to or identi- 
cal with 2-amino-4,7-dihydroxypteridine-6-acetic acid 
(C,H,0,N,). 

As-pergilli 

Yasuyuki Kaneko, /. Agr. Chem. Soc. Japan 31 122 (1957). 

1050 Erythropterin, C.,H;,0-,N-„ deep red crystals from 0.01 N hydro- 

chloric acid. 

OH 

I /OH 

H2N C=C— CH2OH 

I I 
OH OH 

Mycobacterium tuberculosis var. hominis, M. lacticola 
M. O'L. Crowe and A. Walker, Science 110 166 (1949). 
Rudolf Tschesche and Frederic Vester, Chem. Ber. 86 454 
(1953). 

1051 Biopterin, C9H11O3N5, pale yellow crystals, m.p. 250-280° 

(dec), [aln^" -50° (in 0.1 N hydrochloric acid). 

OH 

I XH— CH— CH3 

N 

/ 
H2N 



-^Kl/-\l 



OH OH 



Yeast, Ochromonas malhamensis 

E. L. Patterson, H. P. Broquist, Alberta M. Albrecht, M. H. 
von Saltza and E. L. R. Stokstad, /. Am. Chem. Soc. 77 3167 
(1955). 



Pfizer Handbook of Microbial Metabolites 



566 



1052 V-Compound (8-Ribityl-6-methyl-7-oxylumazme, Compound A), 
Ct.Hi607N4, colorless crystals, m.p. 263° (dec), [ocW 
+4.5° (c 3.3 in water) +11.45° (in 0.1 N sodium hy- 
droxide solution). 



II M 



XH3 



OH 



.CH3 



CH2 

I 
(CHOH)3 

1 
CH2OH 



T J I 

I ^ 

CHo 

I 
(CHOH)3 

I 
CH2OH 



Eremotheciuvt ashbyii 

Toru Masuda, Toyokazu Kishi and Mitsuko Asai, Chem. and 
Pharm. Bull (Japan) 6 291 (1958). (Structure) 

Toru Masuda, Toyokazu Kishi, Mitsuko Asai and Satoru 
Kuwada, ibid. 7 361, 366 (1959). (Synthesis) 

Waher S. McNutt, /. Am. Chem. Soc. 82 217 (1960). 

1053 G-Compound (8-Ribityl-6,7-dimethyllumazine), C13H18O6N4, 
light yellow needles, m.p. 273° (dec), [ocW -164°. 



HN I 



XH3 



CH3 



CH2 

I 
(CHOH)3 

I 
CH2OH 

Eremothecium ashbyii 

1054 Z-3-Oxykynurenine, CHJH10O4N2 

O 

C— CHo— CH— COOH 

NH2 



O" NH, 



567 



Pteridines and Flavines 



was isolated from the same culture. This metabolite re- 
sembles 3-oxyanthranilic acid, known to be a biosynthetic 
precursor of nicotinic acid. 

Toru Masuda, Pharm. Bull. (Japan) 4 71 (1956). (Iso- 
lation) 

Idem., ibid. 5 28 (1957). (Structure) 

Toru Masuda, Toyokazu Kishi, Mitsuko Asai and Satoru 
Kuwada, Chem. and Pharm. Bull. (Japan) 7 361 (1959). 
(Synthesis) 

1055 Rhizopterin (N"'-Formylpteroic Acid) (Streptococcus lactis R 
Factor) (SLR Factor), Ci5Hio04N„, light yellow crystals, 
m.p. >300°. 



OH 






CH.— N— / \— COOH 
CHO 



H..N 



Rhizopus nigricans 

Edward L. Rickes, Louis Chalet and John C. Keresztesy, 
J. Am. Chem. Soc. 69 2749 (1947). 

Donald E. Wolf, R. Christian Anderson, Edward A. Kaczka, 
Stanton A. Harris, Glen E. Arth, Philip L. Southwick, Ralph 
Mozingo and Karl Folkers, ibid. 69 2753 (1947). (Synthesis) 

1056 Riboflavin (Vitamin B^), Ci7H2i,OeN4, yellow-orange micro- 
crystalline powder, m.p. ~280° (rapid heating), [a]ir^ 
-112° to -122° (50 mg. in 2 ml. of 0.1 N alcoholic 
sodium hydroxide diluted to 10 ml. with water). 







CHo 

I 
(CHOH)3 



CH3 



CH3 



CHoOH 

Ascomycetes such as EremotheciuTU ashbyii and Ashbya 
gossypi produce high yields. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
1954 Vol. II, Richard J. Hickey, Production of riboflavin by 
fermentation, Chap. 5, pp. 157-190. (A review) 




Pfizer Handbook of Microbial Metabolites 



568 



1057 Riboflavin-5'-phosphate, C17H21O9N4P, yellow microcrystals. 



HN 



,^^N^^N 




CHo 

H— C— OH 

I 
H— C— OH 

I 
H— C— OH 

I 
CH2 

I 
O 

I 
HO— P— OH 

i 
O 



CH3 



CH3 



Yeast 

Otto Warburg and Walter Christian, Biochem. Z. 254 438 
(1932); 258 496 (1933); 263 228 (1933). (Isolation) 

H. S. Forrest and A. R. Todd, J. Chem. Soc, 3295 (1950). 
( Synthesis ) 

1058 Folic Acid (Pteroyl glutamic Acid Folacin, Vitamin B^.), C19H19- 
OgN-, pale yellow-orange needles, which char above 250°. 



HoN 



OH 



CH2— NH- 



N Y 1' 



(/ ^C— NH— CH— CH2— CH2— COOH 
COOH 



Yeasts and certain higher fungi 

Yields of 19-80 ^g. per gram of dry cell weight are ob- 
tained from brewers' yeast. 

Leland A. Underkofler and Richard J. Hickey, "Industrial 
Fermentations," Chemical Publishing Co., Inc., New York, 
1954 Vol. Ill, J. M. Van Lanen, Production of vitamins other 
than riboflavin. Chap. 6, pp. 191-216. (A review) 



569 Pteridines and Flavines 

1059 Citrovoriim Factor (Folinic Acid-SF, Leucovorin, N^-Formyltet- 
rahydrofolic Acid) C^H.-fi^N- (Trihydrate) : Buff crys- 
tals, m.p. 248-250° (dec), [a],,-" +16.76 (c 3.52 on 
anhydrous basis in 5% sodium bicarbonate solution). 



C— H O COOH 

OH I ^^ II I 

I ,CH..— NH— (/ \)— C— NH— CH 

CH 

I 
H2N H CH2 

1 
COOH 






Yeasts (probably widely distributed) 

The corresponding compound with the formyl group 
transferred to the amine group of the p-aminobenzoic acid 
moiety (Nj,,) is also known. 

C. H. Hill and M. L. Scott, J. Biol. Chem. 196 195 (1952). 
(Isolation from brewers' yeast) 

A. G. M. Sjostrom and L. E. Ericson, Acta Chem. Scand. 7 
870 (1953). (Isolation from eight lichens) 

Donna B. Cosulick, Barbara Roth, James M. Smith, Jr., 
Martin E. Hultquist and Robert P. Parker, /. Am. Chem,. Soc. 
74 3252 ( 1 952 ) . ( Structure ) 

1060 Flavine-Adenine-Dinucleotide, CotH^sOi^NciPo, amorphous white 
powder. 

NH2 



N 






o 

li II 

CHo— O— P— O— P— O— CH, 

1 I 
OH OH 

HC— OH OH OH 

I 
HC— OH 

I 
HC— OH 

I 
CHo 



CH3 I o 

CH, 



Pfizer Handbook of Microbial Metabolites 570 

Yeasts, molds, bacteria (widely distributed) 

Otto Warburg and Walter Christian, Biochem. Z. 298 150 

(1938). (Isolation) 

S. M. H. Christie, G. W. Kenner and A. R. Todd, Nature 170 

924 (1952). 

Idem., J. Chem. Soc, 46 (1954). (Synthesis) 

J. G. MofFatt and H. G. Khorana, /. Am. Chem. Soc. 80 

3756 (1958). (Synthesis) 

1061 Fermentation "Lactobacillus casei" Factor (Teropterin, Pteroyl-y- 
glutamyl-y-glutamylglutamic Acid), C29H33O12N9. 




CH2— NH— <f \— CO— NH— CH— COOH 

{CH,)2 



V '-" 



H2N CO 

NH 
HOOC(CH2)2— CH— NH— CO— (CHo)^— CH— COOH 
COOH 

Corynebacterium sp. 

Brian L. Hutchings, E. L. R. Stokstad, Nestor Bohonos, Na- 
than Sloane and Y. Subbarow, Ann. N. Y. Acad. Sci. 48 265 
(1946). (Isolation) 

J. H. Boothe, J. H. Mowat, B. L. Hutchings, R. B. Angler, 
C. W. Waller, E. L. R. Stokstad, J. Semb, A. L. Gazzola and 
Y. Subbarow, /. Am. Chem. Soc. 70 1099 (1948). 

J. H. Boothe, J. Semb, C. W. Waller, R. B. Angler, J. H. 
Mowat, B. L. Hutchings, E. L. R. Stokstad and Y. Subbarow, 
ibid. 71 2304 (1949). (Synthesis) 

1062 Vitamin Bg Conjugate (Pteroylhexaglutamyl glutamic Acid), 

C4oHoiOo4Ni3. 

The structure is like that of the preceding formula, but 
with four more glutamic acid units in the polypeptide side- 
chain. 

Bacteria, yeasts, molds (widely distributed among 
microorganisms ) 

P. R. Burkholder, Ilda McVeigh and Katherine Wilson, 
Arch. Biochem. 7 287 (1945). 

J. J. Pfiffner, D. G. Calkins, E. S. Bloom and B. L. O'Dell, 
J. Am. Chem. Soc 68 1392 (1946). (Structure) 



571 Pteridines and Fla vines 

1063 Ptcridine pigment. 

A pigment which fluoresces under U.V. light is produced 
by Microsporum species (some of which cause ringworm). 
This pigment has been isolated and purified to some ex- 
tent. The infrared spectrum indicates that it is a pteridine, 
probably trisubstituted, and possibly 2-NHo (or — OH), 
4— OH and 6— CH.OH substituted. 

Microsporuw gijpseum, M. canis 

Frederick T. Wolf, Ernest A. Jones and Helene A. Nathan, 
Nature 182 475 (1958). 



19. 



Unclassified Metabolites 



1064 Aburamycin (M5-18903), yellow crystals, m.p. 163-165° (169- 

, 171°), [a]D'° +24.56° (c 1 in methanol) [aW -29° (c 0.5 
in methanol). 

Absorbs 2 moles of Ho. Acetylates (m.p. acetate = 205- 
207°). A weakly acidic antibiotic, apparent molecular 
weight 1295. Aburamycin and M5-18903 appear to be 
optical antipodes of the same compound. 

Streptomyces spp. 

Haruo Nichimura, Toshiaki Kimura, Katsuya Tawara, Kunio 
Sasaki, Kiyoshi Nakajima, Noboru Shimaoka, Saburo Okamoto, 
Masafumi Shimohira and Jun Isono, J. Antibiotics (Japan) 
lOA 205 (1957). 

Richard M. Gale, Marvin M. Hoehn and Mack H. McCor- 
mick, "Antibiotics Annual 1958-1959," Medical Encyclopedia, 
Inc., New York, p. 489. 

1065 Actinobolin, Cj;iH2o-2206N2, amorphous hygroscopic white pow- 

der, [aW" (Sulfate) 4-54.5° (c 1 in water). 

An amphoteric antibiotic. Forms an acetate: m.p., 
partial m. at 130°, resolidified 145°, dec. 263-266°, [a]^-^ 
+58° (c 1 in water). Positive ninhydrin, ferric chloride, 
KMn04, Tehlings, iodoform tests. Absorbs no hydrogen. 

Streptomyces sp. 

Theodore H. Haskell and Quentin R. Bartz, "Antibiotics An- 
nual 1958-1959," Medical Encyclopedia, Inc., New York, p. 
505. 

1066 Actinoleukin (C9Hi20HN2)n, colorless crystals, m.p. 191° (dec). 

Analysis: C 55.53, H 6.05, N 14.05 
55.68, 5.98, 14.01 
Negative biuret, ninhydrin, Tollens, Fehling. Positive 
FeClg. 



5y3 Unclassified Metabolites 

Streptomyces aureus 

Masahiro Ueda, Yukio Tanigawa, Yoshiro Okami and Hamao 
Umezavva. /. Antibiotics (Japan) 7A 125 (1954). 

1067 Akitamycin, [a],r' +158° (c 0.5 in dimethylformamide), U.V. 

291, 303.5, 319 m^t. Tetraene, C 57.26, H 7.68, N 1.64. 
Streptomyces akitaensis 
J. Antibiotics (Japan) 12B 293, 295, 297 (1959). 

1068 Albidin, C-,H402 (proposed), red needles, not melting below 

380°. 

Unstable. Most stable below pH 3. 

Penicilliurn albidum 

P. J. Curtis and J. F. Grove, Nature 160 574 (1947). 

P. J. Curtis, H. G. Hemming and C. H. Unwin, Brit. Mycol. 
Soc. Trans. 34 332 (1951). 

1069 Albofungin, bright yellow powder, dec. 190°, U.V. 240, 255, 305, 

375 m,jL. Contains C, H, N, O. 

Streptomyces albus var. fungus 

A. S. Chochlov, Czech. Symposium on Antibiotics (Prague), 
154 (1959). 

1070 Albomycetin, C32H54O9N (proposed), colorless crystals, m.p. 

166°. 

A basic substance precipitated by ammonium reineck- 
ate. Positive Fehlings, Tollens, cherry colored Elson- 
Morgan. Negative FeCIa, Sakaguchi, Molisch, Millon. 
May be a macrohde. 

Streptomyces albus 

Bunji Takahashi, J. Antibiotics (Japan) 7A 149 (1954). 

1071 Alboverticillin (Hydrochloride), colorless, amorphous, [a]D^° 

-33.5° (c 1.0 in water). 

Negative U.V., Tollens, Molisch, Benedict, maltol, Elson- 
Morgan, biuret, Millon, Sakaguchi, anthrone and FeClg. 
Positive ninhydrin, Fehling. 

Streptomyces sp. 

Kenji Maeda, Sinichi Kondo, Kofumi Ohl, Hiroko Kondo, 
E. Lin Wang, Yasusuke Osato and Hamao Umezawa, 7- Anti- 
biotics (Japan) HA 30 (1958). 

1072 Aliomycin, yellowish brown powder. Contains C, H, N, O, S. 

Pentaene. U.V. 321, 330, 351 m^x. 

Positive Fehling (on heating, weakly positive Molisch, 
red purple in concentrated H2SO4). 

Streptoinyces acidomyceticus 



Pfizer Handbook o£ Microbial Metabolites 574 

Seizi Igarasi, Koichi Ogata and Akira Miyake, J. Antibiotics 
(Japan) 9B 101 (1956). 

1073 Allomycin,* CogH4409, crystalline, m.p. 237-239° (dec.) [xW 

-118.8 ± 0.5° (c 0.98 in 0.1 N hydrochloric acid). 

Streptomyces sindenensis 

Koichi Nakazawa, Shigehiro Fujii, Michitaka Inoue, Hiroshi 
Hitomi, Ohira Miyake and Jyuzo Kaneko, /. Antibiotics (Ja- 
pan) 7B 168 (1954). 

Sueo Tatsuoka, Koichi Nakazawa, Michitaka Inoue and 
Shigehiro Fujii, J. Pharm. Soc. Japan 75 1206 (1955). 

1074 Alternarine, colorless needles, m.p. 230°. 

Alternaria solani 

Herman Darpoux, Albert Faivre-Amiot and Louis Roux, 
Compt. rend. 2:J0 993 (1950). 

1075 Althiomycin, Ci-Hi^NjS.O,., colorless crystals, m.p. 172-174° 

(dec.) (browning" from 120-160°), [7.W +20.3 (c 1.33 
in methyl cellosolve). 

Unstable at pH <5.0 or >7.0. 

A streptomycete 

Hiroshi Yamaguchi, Yuya Nakayama, Keiichi Takeda, 
Kosaku Tawara, Kenji Maeda, Tomio Takeuchi and Hamao 
Umezawa, /. Antibiotics (Japan) lOA 195 (1957). 

1076 Anisomycin (PA-106, PA-107), Ci4Hi,,04N, white needles, m.p. 

140°, [a]D'' -45° ± 3° (c 1.0 in chloroform). 

Streptomyces griseolus, other Streptomyces spp. 

Ben A. Sobin and Fred W. Tanner, Jr., J. Am. Chem. Soc. 76 
4053 (1954). 

Fred W. Tanner, Jr., B. A. Sobin and J. F. Gardocki, "Anti- 
biotics Annual 1954-1955" Medical Encyclopedia, Inc., New 
York, p. 809. 

1077 Antibiotic A 246,t C4]H,;6.-„0]4, crystalline, m.p. 235° (dec), 

[aW -160° (c 0.2 in methanol). 

Reacts with HIO4. 

Streptomyces sp. 

M. L. Dhar, V. Thaller and M. C. Whiting, Proc. Chem. Soc, 
148 (1958). 

1078 Antibiotic B-456, m.p. 176° (dec), UW' -22.9°. 

C 57.52, H 6.67, N 11.12 

Positive biuret, Millon. Negative Molisch, Benedict, 
Fehling. 

* See amicetin. 

t Identical with lagosin, entry 229. 



575 Unclassified Metabolites 

Valine, leucine, proline, aspartic, glutamic, D-tyrosine 
and ornithine produced after hydrolysis. 
Bacillus suhtilis 
Yuzuru Tanaka, J. Antibiotics (Japan) 9B 1 (1956). 

1079 Antibiotic C-159. 

U.V. 260-280, 345 m^x in aqueous solution. 
C 58.7, H 7.4, N 9.9, O 24.0 

Inhibits growth of organisms containing glycine, ala- 
nine, threonine, aspartic acid. 
Streptojiiyces caiius 
Bristol Laboratories, British Patent 814,794 (1959). 

1080 Antibiotic D-13, dense crystals, m.p. 243°. 

C 56.91, H 6.97, O 22.61, N 13.51. 

Streptomyces vinaceus-drappiis 

Upjohn Co., British Patent 708,686 (1954). 

1081 Antibiotic E-212, colorless needles, m.p. 233-234°. 

U.V. 235, 273 m/x in 0.1 N hydrochloric acid. C 49.14, 
H 4.34, N 23.77, O 22.55 

Negative ninhydrin, biuret, Fehling, FeClj, Molisch, 
Millon and Ehrlich. 

Streptomyces sp. like S. albus 

Ko Kikuchi, J. Antibiotics (Japan) 8A 145 (1955). 

1082 Antibiotic LA-7017, greenish yellow powder, m.p. 154-157° 

(dec), [aj,r' —155° (c 0.4 in ethanol). 

Contains only C, H, O (C 56.99, H 7.18). Contains 
two acidic groups, Equiv. Wt. = 1180. Decolorized 
KMn04. Negative Fehling's test. 

Streptomyces sp. 7017 

P. Sensi, A. M. Greco and H. Pagani, Antibiotics and Chemo- 
therapij 8 241 (1958). 

1083 Antibiotic M-4209, C4„-4i;Hfi-.7,0,,;N, white crystals m.p. 210- 

214° (dec), [a],r' -54 ± 2° (c 1 in methanol), U.V. 240, 
330 ruii. 

Methoxyl, acetyl and iso-valeryl groups present. 

Streptomyces hygroscopicus 

James D. Dutcher, John Vandeputte, Sidney Fox and L. J. 
Heuser, Antibiotics and Chemotherapy 3 910 (1953). 

1084 Antibiotic WC 3628, C4.H7..iO,fiN, white crystals, m.p. 220-222° 

(Kofler), [a]D"-57±3° (c 0.5 in ethanol). 



Pfizer Handbook of Microbial Metabolites 576 

Streptomyces sp. WC 3628 

McCormick, Canadian Patent 513,324 (1955). 

1085 Antibiotic T, trichothecin-like, crystalline prisms, m.p. 126°, 

[ale'" +135° (c 1 in chloroform). 

A basidiomycete. 

E. T. Glaz, Eszter Scheiber, J. Gyimesi, I. Horwath, Katalin 
Steczek, A. Szentirmai and G. Bohus, Nature 184 908 (1959). 

1086 Antibiotic X-206, C4fjHgoOi3, colorless crystals, m.p. 126-128°, 

[a]D-" +15.0° (c 2.0 in methanol). 

Streptomyces sp. 

Julius Berger, A. I. Rachlin, W. E. Scott, L. H. Sternbach 
and M. W. Goldberg, J. Am. Chem. Soc. 72 5295 (1951). 

1087 Antibiotic X-464, C25H40O7, white crystals, m.p. 172-174° 

(dec), [aW +65.9° (c 2.0 in methanol). 

Streptomyces sp. 

Julius Berger, A. I. Rachlin, W. E. Scott, L. H. Sternbach 
and M. W. Goldberg, J. Am. Chem. Soc. 73 5295 (1951). 

1088 Antibiotic X-537A, C34H-,o08, colorless crystals, m.p. 100-109°, 

[alD'** -7.2° (c 1.0 in alcohol), U.V. 317, 249 m^ in iso- 
propyl alcohol. 

Positive FeClg test. 

Streptomyces sp. 

Julius Berger, A. I. Rachlin, W. E. Scott, L. H. Sternbach 
and M. W. Goldberg, /. Am. Chem. Soc. 73 5295 (1951). 

1089 Antibiotic X-1008, CaoHgsOyNgS, cube-like crystals, m.p. 209- 

216° (dec), [o^W -282° (c 1 in chloroform). 

Resembles echinomycin 

Streptomyces sp. 

J. Berger, E. R. LaSala, W. E. Scott, B. R. Meltsner, L. H. 
Sternbach, S. Kaiser, S. Teitel, E. Mack and M. W. Goldberg, 
Experientia 13 434 (1957). 

1090 Antibiotic from B. cepae, colorless crystals, m.p. 185° (dec.) 

C 40.8, H 5.3. 

Bacillus cepae 

Isolated from rotting onion. 

M. Fiuczek, Med. Doswiadczalna i Mikrobiol. 2 175 (1950). 
(Biol. Abstr. 26 3975). 

1091 Antibiotic from B. pumilis, CgHgNoOaS, white crystals, m.p. 

252°. 

Negative ninhydrin. 



577 



Unclassified Metabolites 



Bacillus pumilis 

A. T. Fuller, Nature 175 722 (1955). 

1092 Antibiotic from Monosporium honorden, CiyHibOy, colorless 

crystals, m.p. 193.5°, [a],r" +203° (in chloroform). 

Two phenolic hydroxyl groups, one active hydrogen on 
an aromatic ring, one double link in a side-chain and a 
free carboxyl group present. 

Molecular structure may be closely related to the struc- 
ture proposed for mycophenolic acid. 

Monosporium honorden 

P. Delmotte and J. Delmotte-Plaquee, Nature 171 344 
(1953). 

1093 Antibiotic from Penicillium spinulosum, fine white needles, 

m.p. 183-185°. 

Penicillium spinulosum 

Shegejii Kondo and Bunji Takahashi, /. Penicillin (Japan), 
1 147 (1947). 

1094 Antibiotic from S. abikoensis, yellow powder. Heptaene. U.V. 

242, 358, 400 m/x in ethanol. Actinoleukin in mycelium. 

Streptomyces abikoensis 

Masahiro Ueda and Hamao Umezawa, J. Antibiotics (Ja- 
pan) 9A 86 (1956). 

1095 Antibiotic from S. /ungicirfict/s. U.V. 290, 303, 317 m^^. Similar 

to fungicidin or rimocidin. 

Positive Fehling, Molisch, Negative Millon, Sakaguchi, 
Schiff, ToUens, FeCl.v Blue with FeClg-K ferricyanate; 
decolorizes KMn04. 

Streptomyces fungicidicus 

Hamao Umesawa, Yoshio Okami and Ryozo Utahara, Japa- 
nese Patent 5744 (1956). 

1096 Antibiotic from S. griseus. Heptaene. U.V. 359-362, 378- 

382, 401-405 m/x. 

Streptomyces griseus 

Richard A. Pledger and Hubert Lechevalier, "Antibiotics 
Annual 1955-1956," Medical Encyclopedia, Inc., New York, 
p. 249. 

1097 Antibiotic 26/1, yellow crystalline. Heptaene. U.V. 359, 380, 

404 m^ in ethanol. 

Alcohol solution turns violet with H2SO4; decolorizes 
KMn04. Negative biuret and ninhydrin. 



Pfizer Handbook of Microbial Metabolites 578 

Actinomyces globisporus 

V. A. Tsyganov, P. N. Golyakov, A. M. Bezborodov, V. P. 
Namestnikova, G. V. Khopko, S. N. Solov'ev, M. A. Malyshkina 
and L. O. Bol'shakova, Antibiotiki 4 21 (1959). 

1098 Antibiotic 446, white crystalline powder, m.p. 81-87°, [a]u^^ 

-82° (c 0.5 in ethanol). U.V. 230-231, 280 m^i. 

C 60.47, H 7.99, N 2.02 

Negative Fehling. 

Nocardia mesenterica 

Masahiro Ueda and Hamao Umezawa, J. Antibiotics (Ja- 
pan) 8A 164 (1955). 

1099 Antibiotic 720-A,* C2SH40O9N0, white needles, m.p. 139.5-140°, 

[a]i/' +73.5° (c 1.0 in acetone) U.V. 227, 346 m^. 

Positive FeClg; negative Molisch, ninhydrin, biuret, 
Ehrlich and 2:4 DNPH. 

Streptomyces n. sp. 

Yoshio Sakagami, Setsuo Takeuchi, Hiroshi Yonehara, 
Heiichi Sakai and Matao Takashima, /. Antibiotics (Japan) 
9A 1 (1956). 

1100 Antibiotic 587/13, Hydrochloride 

C 39.5, H 6.97, N 15.7, CI 16.75 

Streptomyces lavendulae 

D. M. Trakhtenberg, V. M. Baikina, E. 1. Rodionovskaya, 
1. M. Prosnyakova, O. A. Kalinovskii, Yu V. Zakharova and 
A. A. Khokhlov, Antibiotiki (U.S.S.R.) 4 9 (1959). 

1101 Antibiotic 1037, crystalline needles, m.p. 283-289°, [^W^ -51°. 

C 49.33-49.47, H 4.56-4.90, N 23.75-24.14, no halo- 
gen or sulfur 

Streptomyces sp. 

Hiroshi Yamamoto, Shlgehlro Fujli, Koichi Nakazawa, 
Akira Miyake, Hiromu Hitomi and Masahiko Imanishl, Ann. 
Repts. Takeda Research Lab. 16 26 (1957). 

1102 Antibiotic 6270,t CooHs-NeSO^.^, crystalUne. 

Streptomyces fiavochromo genes 

M. G. Brazhnikova, Czech. Symposium on Antibiotics 
(Prague), 140 (1959). 

1103 Antibiotic 6706,1 C26-27H32O8N4, colorless needles, m.p. 214- 

216°, U.V. 304" ni;,."" 

Gives negative FeCLj, Fehling, Tollens, ninhydrin and 

Millon tests. 
* See entry 269 (antimycin Ai). 
t Cf. entry 1089. 
X See pyridomycin, entry 752. 



579 Unclassified Metabolites 

Streptomyces sp. 

Masahiko Kuraya, Bunji Takahashi, Yorio Hinuma, Takaaki 
Yashima, Kenzo Watanabe, Masa Kuroya and Susumu Ha- 
mada, /. Antibiotics (Japan) 7A 58 (1954). 

1104 Antifungal Substance, colorless needles, m.p. 283-289°, [a]i,'' 

-51°. 

A water-soluble compound similar to toyokamycin and 
monilin. Analysis: C 49.33-49.47, H 4.56-4.90, N 
23.75-24.14. 

Streptomyces sp. 

Hiroshi Yamamoto, Shigehiro Fujii, Koichi Nakazawa, Akira 
Miyake, Hiromu Hitomi and Masahiko Imanishi, Takeda 
Kenkyusho Nempo 16 26 (1957). 

1105 Antifungal substance produced by Streptomyces strain No. 1037. 

Crystalline needles, m.p. 283-289°, [alo'-' -51°. C 
49.33-49.47 H 4.56-4.90 N 23.75-24.14, no halogen or 
sulfur. 

It seems to belong to the same group of substances as 
toyokamycin and monilin. 

Hiroshi Yamamoto, Shigehiro Fujii, Koichi Nakazawa, Akira 
Miyake, Hiromu Hitomi and Masahiko Imanishi, Ann. Rept. 
Takeda Research Lab. 16 26 (1957). 

1106 Argomycin, ConH^gO^N, m.p. 164°, [(x]ir' +8.2° (in ethanol). 

May be a macrolide. 

Streptomyces griseolus 

Toji Hata, Yoshimoto Sano, Hideo Tatsuta, Ryozo Sugawara, 
Akihiro Matsumae and Kokichi Kanamori, J. Antibiotics (Ja- 
pan) 8A 9 (1955). 

1107 Aspelein, C29H20O10, dark red plates, no m.p. 

This pigment contained two hydroxyl groups (diacetate, 
yellow crystals, m.p. 276-285°) and an alkoxyl group. 
Spectra described. 

Aspergillus elegans 

P. E. Gregoire, Bull. soc. chim. biol. 33 1681 (1951). 

Aterrimins — complex containing aterrimins A and B, exhibiting 
characteristics of a lactone; contain C, H and O and have 
no definite m.p. 

1108 Aterrimin A, [a],.-" +245° in ethanol. U.V. 277, 287, 310-325 

m^ in absolute alcohol C. 65.5 H 7.8 O 26.7 (by differ- 
ence). 



Pfizer Handbook of Microbial Metabolites 580 

1109 Aterrimin B, [aU-° +342° in ethanol. U.V. same as A. C 69.7 

H 8.05 O 22.25 (by difference). 

Bacillus subtilis var. aterrimus 

Gordon Alderton and Neva S. Snell, U. S. Patent 2,850,427 
(1958). 

1110 Aureolic Acid, Mg salt: (C56.6oH96-io4029-3i)2Mg, yellow crystals, 

[aln +68° (c 1 in methanol). 

A weak acid, green FeClg test, negative FehUngs, 
an throne. 

Streptomyces sp. 

Walton E. Grundy, Alma W. Goldstein, Charles J. Rickher, 
Marjorie E. Hanes, Halleck B. Warren, Jr. and John C. Sylves- 
ter, Antibiotics and Chemotherapy 3 1215 (1953). 

im Azalomycin B, C14H24O5, white needles, 185-187° (dec.) [ajc^' 
-48° (c 1.0 in methanol). U.V. 252.5 m,x. 
Streptomyces hygroscopicus 
Manoru Aral, /. Antibiotics (Japan) ISA 51 (1960). 

1112 Azalomycin F, C30H50O10N2, white needles, m.p. 125-127° (dec.) 

[ccW +46° (c 1.0 in methanol). 

U.V. resembles that of musarin and hygrostatin. I.R. 
differs. 

Positive ninhydrin, negative FeCls, MoHsch, anthrone 
and Millon. 

Streptomyces hygroscopicus 

H. D. Tresner and E. J. Backus, Appl. Microbiol. 4 243 
(1956). 

S. A. Waksman and A. T. Henrici, Bergey's "Manual of 
Determinative Bacteriology," 1957, pp. 796-797. 

Mamoru Arai, /. Antibiotics (Japan) ISA 51 (1960). 

1113 Baccatine A, C26H48O6N0 (proposed). Colorless crystals, m.p. 

135°. Mol. Wt. -^480. 

May be a depsipeptide (peptolide). 

Gibberella baccata. 

Jean Guerillot-Vinet, A. Guerillot-Vinet, Lucien Guyot, Jac- 
ques Montegut and Louis Roux, Compt. rend. 230 1424 (1950). 

M. M. Shemyakin, Angew. Chem. 72 342 (1960). 

1114 Bacilipin A, sheaves of needles, m.p. 76-78°. 

C 42.6, H 6.3, N 2.5, Ba 24.6. 
Negative Mohsch, 2,4-DNPH, AgNOg. 
Positive Br2. 



581 Unclassified Metabojites 

M15 Bacilipin B, crystals, m.p. 105°. 

C 52.45, H 6.75, N 2.09, Ba 21.6 

Gave same tests as Bacilipin A. 

Both A and B gave positive ninhydrin after hydrolysis. 

Bacillus subtilis 

G. G. F. Newton, Brit. J. Exptl. Biol. -iO 306 (1949). 

1116 Bacilysin, white powder containing C, H, O and N. 

Gives a positive ninhydrin; negative biuret and MoUsch 
tests. 

Produced by the soil bacillus NTCC 7197. 

E. P. Abraham and H. W. Florey, "Antibiotics," Vol. I 
Antibiotics from bacteria in the genus bacillus, Oxford Uni- 
versity Press, London, 1949 Chap. 10, pp. 457-458. 

1117 Biformyne 1 (Biformin), C9H20O0, white crystalHne solid, m.p. 

40-43°, U.V. 276, 278, 291 m^x in alkaH. 

Polyporus biformis 

Marjorie Anchel and Marvin P. Cohen, /. Biol. Chem. 208 
319 (1954). 

1118 Blasticidin A, C4r,..,2, H^.^o N4.7, light yellow powder, m.p. 197- 

201°. U.V. 216 m^. Soluble in HoO. 

1119 Blasticidin B, colorless liquid, b.p. 36° (0.001 mm.). Insoluble 

in H.O. 

1120 Blasticidin C, red-brown powder. Insoluble in HoO. 

Streptomyces griseochromogenes 

Kazuo Fukunaga, Tomomasa Misato, Itaru Iskii and Masaru 
Asakawa, Bull. Agr. Chem. Soc. (Japan) 19 181 (1955). 

1121 Blasticidin-S, Ci4H2o05N6, white needles, m.p. 235° (dec), 

[ale" +108.4° (c 1.0 in water). 

A basic antibiotic (forms a picrate). 

Negative FeCl.j, Fehling, Tollens, Millon, Ehrlich, 
Sakaguchi, MoUsch, biuret, ninhydrin, aldehyde and 
ammoniacal AgNOg tests. Blasticidin-S is a member of a 
complex with at least three other components, blasticidins 
A, B and C. 

Streptomyces griseochromogenes 

Setsuo Takeuchi, Kosei Hirayama, Kazaburo Ueda, Heiichi 
Sakai and Hiroshi Yonehara, /. Antibiotics (Japan) llA 1 
(1958). 



Pfizer Handbook of Microbial Metabolites 582 

1122 Borrelidin, C2sH4:{0(jN, m.p. 145°, [a],r' -28° (in ethanol). 

An acidic compound. 
Streptomyces rochei 

J. Berger, L. M. Jampolsky and M. W. Goldberg, Arch. 
Biochem. 22 476 (1949). 

1123 Caerulomycin, Ci2Hi,02N;^, colorless needles, m.p. 175°. 

Red FeCl^ test. Contains one methoxyl group. 

Streptomyces caeruleus 

A. Funk and P. V. Divekar, Can. J. Microbiol. 5 317 (1959). 

1124 Camphomycin, white needles, m.p. -^ 149°. Positive Nessler and 

Tollens. 

Streptomyces rutgersensis var. castelarense 
Augusto P. Cercos, Rev. argentian agron. 20 53 (1953). 

1125 Candidulin, C,,H,,-,0..jN, white needles, m.p. 88°, [a]ir' +15° 

±2° (c 1 in chloroform). 

A neutral, non-aromatic substance. Negative ninhydrin, 
2,4-DNPH, FeCl-i. 

Aspergillus Candidas 

P. G. Stansly and N. H. Ananenko, Arch. Biochem. 23 256 
(1949). 

1126 Canescin, Cir,Hi,0-, white needles, m.p. 201-202° (dec). 

Purple color with FeClj in ethanol. 
Penicillium canescens 
Yield 30-110 mg. per liter. 

P. W. Brian, H. G. Hemming, J. S. Moffatt and C. H. Unwin, 
Trans. Brit. Mtjcol. Soc. 36 243 (1953). 

1127 Cardinophyllin (Carzinophilin), potassium salt: colorless 

needles, m.p. 220° (dec). 

Contains C, H, O, N. Positive xanthoprotein, negative 
ninhydrin, diphenylamine. Negative resorcinol, Millon, 
Liebermahn. 

Streptomyces sahachiroi 

Toju Hata, Fumiwaka Koga, Yoshimoto Sano, Koklchi 
Kanamori, Akihiro Matsumae, Ryozo Sugawara, Tadashi Hoshi 
and Tatsuo Shima, /. Antibiotics (Japan) 7A 107 (1954). 

Fujiki Hata and Takamoto Sano, Japanese Patent 7590 
(1956). 

1128 Carzinophilin A, colorless needles, m.p. 217-222° (dec), [cz]i."^ 

+57.8° (in chloroform). 

Positive ninhydrin, 2,4-DNPH, bromine uptake, an- 



583 Unclassified Metabolites 

throne, Baeyer, xanthoproteic. Unstable in aqueous solu- 
tion. 

Strcptomijces sahachiroi n. sp. 

Hideo Kamada, Shigetoshi Wakaki, Yasuo Fujimoto, Keitaro 
Tomioka, Satoshi Ueyama, Hakudai Marumo and Keizo Uzu, 
;. Aiitibiotics (Japan) 8A 187 (1955). 

1129 Cerevioccidin, C:.._.H:^n04Nr„ colorless needles, m.p. 249° (dec). 

Negative biuret, ninhydrin, Fehling, Sakaguchi, Tollens, 

glucosamine. Positive Janovsky. 

Streptoviyces sp. resembling S. cacaoi 

Satoru Yamashita, Teruzo Sawazaki, Makoto Kawasaki, 

Goto Nakamura, Kentaro Anzai, Kiyoshi Isono, Yoshiko 

Serizawa, Yoshiko Sekiyama and Saburo Suzuki, /. Antibiotics 

(Japan) 8A 42 (1955). 

Chlamydosporin, complex of two closely related antibiotics pro- 
duced by the fungus Fiisarium MLF 1230 found in insects 
and their larvae. 

1130 Chlamydosporin A, light brown amorphous substance insoluble 

in water. 

1131 Chlamydosporin B, colorless, crystalline, soluble in water. 

Both contain 4.3% N but no sulfur. 

Albert Faivre-Amiot, Hermon Darpoux and Louis Roux, 
Compt. rend. 235 912, 982 (1952). 

1132 Chromomycin A.j (main component of complex), C.22.-2Aiir2.:MOii, 

yellow powder, m.p. 183° (dec), [a],.'-'" -26° (c 1 in 
ethanol. 

May be related to the actinomycins. 

Streptomyces griseus No. 7 

Yoshitomo Aramaki, Junmei Watanabe, Ichiro Ishikawa, 
Akira Miyake, Homu Ito, Koichi Nakazawa, Koichi Ogata, 
Motoo Shibata, Masaji Igarashi and Kazuo Tanabe, Ann. Repts. 
Takeda Research Lah. 14 60 (1955). 

Tatsuoka et al, Gann. 49 Suppl. 23 (1958). 

S. Wakaki et al.. Antibiotics and Chemotherapy 8 228 
(1958). 

Motoo Shibata, Kazuo Tanabe, Yoshio Hamada, Koiti 
Nakazawa, Akira Miyake, Hiroshi Hitoma, Masuo Miyamoto 
and Komei Mizuno, /. Antibiotics (Japan) 13B 1 (1960). 

1133 Chrysergonic Acid, C.^H;,, •j^O,.,, fine yellow needles, m.p. 268— 

270° from chloroform (250-257° from acetic acid), UW" 
-3° -^ +34° (in pyridine). 
Claviceps purpurea 



Pfizer Handbook of Microbial Metabolites 584 

A. Stoll, J. Renz and A. Brack, Helv. Chim. Acta 35 2022 
(1952). 

1134 Chrysomycin, C22H00O7 (proposed), greenish yellow crystals, 

m.p. 255-260° (dec), [o(W^ +16° (c 1 in acetic acid). 

Neutral, photosensitive compound. Takes up 4H2 with 
loss of color. 

Streptomyces sp. 

Frieda Strelitz, Helen Flon and Igor V. Asheshov, J. Bacteriol. 
69 280 (1955). 

1135 Clitocybin, colorless crystals, m.p. 77°. 

Clitocybe Candida 

A. Charles Hollande, Compt. rend. 221 361 (1945); 228 1758 
(1949). 

1136 Coelicolorin, purpUsh red powder, 142-146°. 

Streptomyces coelicolor 

Yuichi Hatsuta, /. Antibiotics (Japan) 2 276 (1949). 

1137 Collinomycin, orange prisms, m.p. 280°. 

Streptomyces collinus (mycelium) 

Hans Brockmann and Karl-Heinz Renneberg, Naturwissen- 
schaften 40 166 (1953). 

1138 Compound CuHooOgNo. 

A basic red pigment, yellow in alkaline, red in acid 
solutions. Positive Bayer, diazo tests. 
Inocybe patoullardii Bres. 
Helmut Miiller, Dissertation, Wiirzburg, 1959. 

1139 Cosynthetic Factor-1 C14.15H17O7N3, crystalline. An acidic com- 

pound, Mol. Wt. 340-360. 

Thought to be a cefaclor in the biosynthesis of tetra- 
cyclines. 

Streptomyces aureofaciens strain W-5, S. albo-niger, S. 
griseus, S. albas, S. platensis, S. hygroscopicus, S. rimosus 

Jerry Robert Daniel McCormick, Nancy Hazlett Arnold, 
Ursula Hirsch, Philip Andrew Miller and Newell Oscar 
Sjolander, Union of South Africa Patent Application 59-2174 
(1959). 

1140 Croceomycin, C.oRx&Of.,, m.p. 325° (subl. 240° at 1-2 mm.), 

UW^ -32 ±4°. 

Forms a triacetate. Diazomethane adds two methyl 
groups. 

Streptomyces arabicus 



585 Unclassified Metabolites 

Motoo Shibata, Koichi Nakazawa, Akira Miyake, Michitaka 
Inoue and Akira Akabori, Takeda Kenkyusho Nempo 16 32 
(1957). (Chem. Ahstr. 52 10279e) 

1141 Cyanomycin, C15H10N0O0 (proposed), dark blue needles, m.p. 

128° (dec.)- 

A basic antibiotic pigment with pH-indicating prop- 
erties, apparently distinct from other known pigments. 
Aureothricin occurs in the same culture. 

Streptomyces strain No. 4738 

Masanao Funaki, Fumiyasu Tsuchiya, Kiyoharu Maeda and 
Takeshi Kamiya, /. Antibiotics (Japan) llA 143 (1958). 

1142 Datemycin, C-,><Hlo-O^JN4, colorless powder, m.p. 197° (dec), 

[a]i/^ -43.7° (c 1 in water). 

U.V. maximum at 247 m/x. Negative ninhydrin 

6N HCl 

— — > positive ninhydrin. Negative Hopkins- 
Cole, xanthoprotein, Sakaguchi, Millon, Elson-Morgan, 
Molisch, Fehling, silver mirror tests. 

"M-14" strains 

Masahiko Kuroya and Yasuo Koyama, Japanese Patent 6648 
(1959). 

1143 Desertomycin, C33H(;,,.yoOi4N, snow white crystals, m.p. 189°. 

Positive ninhydrin, C-methyl. Acetylates, hydrogenates, 
decolorizes bromine or permanganate. 

A crystalline antifungal agent, flavofungin, has been 
isolated from the same culture. 

Streptomyces flavofungini 

J. Ori and I. Behesi, Nature 181 908 (1958). 

J. tJri, R. Bognar and B. Varga, ibid. 182 401 (1958). 

1144 Diaporthin, C13HJ4O3, white crystals, m.p. 91.5-92.5° [ajn^^ +58° 

(c 1 in chloroform). 
Endothia parasitica 
A. Neelameghan, Hindustan Antibiotics 2 13 (1959). 

1145 Diplococcin, antibacterial substance elaborated by certain milk 

streptococci. In the same category as the sulfur-free 
polypeptides, gramicidin and tyrocidine. Unlike these 
polypeptides diplococcin contains arginine residues, shows 
no tendency to crystallize and is obviously of greater mo- 
lecular complexity. 

C 50.5, H 7.3, N 13.1, no sulfur. 



Pfizer Handbook of Microbial Metabolites 586 

Streptomyces lactis 

A. E. Oxford, Biochem. 7. 38 178 (1944). 

Idem., ibid. 39 xiii (1945). 

1146 Distamycin A, pure white powder, basic, forms salts. 

Positive biuret test. 

CIBA, Australian Patent 28,469 (1957). 

1147 D-Substance, white needles, m.p. 124-125°. 

Highly toxic. 

Streptomyces flavus 0-2 

Isao Takahashi, /. Antibiotics (Japan) 6A 117 (1953). 

1148 Elaiophylin (C^HioOo),,, no N, S, X, white crystals, m.p. 178- 

183° (dec.) [air'' -49° (in chloroform). 

Streptomyces melanosporus (sine melanosporofaciens) 
F. M. Arcamone, C. Bertazzoh, M. Ghione and T. Scotti, 

Giorn. microbiol. 7 207 (1959). 

1149 Endosubtilysin, yellow powder, soluble in alcohol and chloro- 

form. Forms a water-soluble sodium salt. Appears to 
be an organic acid. 

Bacillus subtilis 

Louis de Saint-Rat and Henry R. Olivier, Compt. rend. 222 
297 (1946). 

1150 Enteromycin, m.p. 167-168°, U.V. 282 m,x. 

C 38.2, H 4.62, N 4.3. 

Streptococcus albireticuli 

Teisuke Osato, Masahiro Ueda, Setsuko Fukuyama, Koki 
Yagishita, Yoshiro Okami and Hamao Umezawa, /. Antibiotics 
(Japan) 8A 105 (1955). 

1151 Ergochrysin, CosHosOjo, yellow-golden leaflets, m.p. 266° from 

chloroform (242-244° from alcohol-pyridine). 
Claviceps purpurea 

C. Jacoby, Arch. exp. Pathol. Pharmakol. 39 85 (1897). 
Werner Bergmann, Ber. 65 1486, 1489 (1932). 

1152 Ergoflavin, C3oH2fjO,4, yellow needles, m.p. 350° (dec.) from 

methanol or dioxane, [a]i,-' +37.5° (c 1.236 in acetone). 
Structural features determined : 
4 phenolic hydroxyls 
2 alcoholic hydroxyls 
2 carbonyls 
2 y-lactones 
Claviceps purpurea 



587 Unclassified Metabolites 

The yield is 1-2% of the weight of the dry sclerotia. 

G. Eglinton, F. E. King, G. Lloyd, J. W. Loder, J. R. Marshall, 
Alexander Robertson and W. B. Whalley, /. Chem. Soc, 1833 
(1958). 

The relationship of ergofiavin to the other yellow pig- 
ments, secalonic acid, ergochrysin, chrysergonic acid, 
sclererythrin, scleroxanthin, sclerocristallin and ergoxan- 
thin (some of them identical) is discussed in the above 
paper as well as in an earlier paper by: 

Albert Freeborn, Pharm. J. 88 568 (1912). 

A. Stoll, J. Renz and A. Brack, Helv. Chim. Acta. 35 2022 
(1952). 

1153 Estin, C„iHi40,.CL, m.p. 223°. 

Contains two methoxyl groups. A second and similar 
compound, Nordin, is produced with it. A 143 mg. sam- 
ple of the mixture was obtained from 1480 ml. of culture 
solution. 

Penicillium paxilli var. echinulatum 

Eitaro Komatsu, Japanese Patent 4799 (1953). 

1154 Eumycetin, fine white needles, m.p. 148-150°. 

Positive FeCl.-,, negative biuret, ninhydrin, Molisch, 

Fehling. 

Streptomyces sp. similar to S. purpurochromogenes 
Edwin A. Johnson and Kenneth L. Burdon, J. Bacteriol. 51 

591 (1946). 

1155 Eumycin, amorphous precipitate, heat-stable in acid, unstable 

in alkaline solutions above pH 8.0. 

Bacillus subtilis 

Edwin A. Johnson and Kenneth L. Burdon, J. Bacteriol. 51 
591 (1946). 

1156 Exfoliatin, CotH^oOisCI, colorless needles, m.p. 172°. 

Positive FeClg, Molisch, Negative Fehling. 
Streptomyces exfoliatus 

Hamao Umezawa, Kiyoshi Oikawa and Motoko Suzuki, /. 
Antibiotics (Japan) 5 466 (1952). 

1157 Fairodin, crystalline, m.p. 237-239° (dec.) [a]v'^ -102° (c 1 

in water). 

C 59.6, H 6.7, N 14.3. 
Bacillus brevis 



Pfizer Handbook of Microbial Metabolites 588 

S. Oya, Japanese Patent Application SHO 32-3997 (1957). 

1158 Fermicidin, C14H01O4N, colorless needles, m.p. 96°, [aW^ +52.3° 

±1.5° (c 0.65 in water). 

Streptomyces sp. similar to S. griseolus 

Seizi Igarasi and Shozo Wada, /. Antibiotics (Japan) 7B 

221 (1954). 

1159 Fermizin, C14H21O4N, needles, m.p. 96-98°. 

An antifungal agent. 

Streptomyces griseus 

About 10 g. were obtained from 100 1. of fermentation 
broth. Apparently identical with fermicidin. 

Koichi Ogata, Masaji Igarashi and Shozo Wada, Japanese 
Patent Application 6150 (1958). 

1160 Fervenulin, C7H7O2N5, yellow crystals, m.p. 178-179° (dec). 

Mol. Wt. 193. Acid-stable, base-labile. U.V. peaks at 
275, 239 rrifi. 

Streptomyces fervens 

T. E. Eble, E. C. Olson, C. M. Large and J. W. Shell, 7th 
Annual Symposium on Antibiotics, Washington, D. C, 1959. 

1161 Flavensomycin, pale yellow crystals, m.p. 152°. 

A water soluble compound containing nitrogen but not 
sulfur or halogen. Some carbohydrates tests were posi- 
tive. U.V. maximum at 251 m^u. 
Streptomyces tanaschiensis type 
R. Craveri and G. Giolitti, Nature 179 1307 (1957). 

1162 Flavucidin, C34H55NO9, colorless needles, m.p. 144-145°, [ixW° 

94°, U.V. 275 nifx. 

Positive Molisch. Negative ninhidrin. 

Streptomyces sp. No. 14420 

Motoo Shibata, Koichi Nakazawa, Akira Miyake, Michitaka 
Inoue, Jiro Terumichi and Hiroshi Kawashima, Ann. Rept. 
Takeda Research Lab. 17 16 (1958). 

1163 Folimycin, m.p. 163-164° (dec.) agricultural antifungal anti- 

biotic. 

Streptomyces nayagawaensis n. sp. 

Hiroichi Yamamoto, Koiti Nakazawa, Satoshi Horii and 
Akira Miyake, J. Agr. Chem. Soc. Japan 34 268 (1960). 

1164 Fomecin A, CgHgOg, m.p.: dec. >160°. 

Weakly acidic, thermostable, alkali labile. 
Fomes (Polyporus) juniperinus 



589 Unclassified Metabolites 

Marjorie Anchel, Annette Hervey and William J. Robbins, 
Proc. Nat. Acad. Sci. U. S. 38 655 (1952). 

1165 Fuscomycin, m.p. 180° (dec.)- 

Streptomyces fuscus 

Fujiki Hata and Keigen Sano, Japanese Patent 5046 (1953). 

1166 Glutinosin, C4sH,ioOio (proposed), colorless plates, gradual dec. 

to 300°, [aW" ^+54° (c 0.2 in benzene). 
Metarrhizium gliitinosum 
P. W. Brian and J. C. McGowan, Nature 157 334 (1946). 

1167 Grisaminc, C^sH^sOioN^ or Co„H;5,)07N4 (proposed), colorless 

needles, m.p. 167-170°. 

Negative Fehling, FeCIs, Sakaguchi, ninhydrin, biuret. 

Streptomyces sp. similar to S. griseoflavus 

Teruzo Sawazaki, Goto Nakamura, Makato Kawasaki, Satoru 

Yamashita, Kiyoshi Isono, Kentaro Anzai, Yoshiko Serizawa, 

Yoshiko Sekiyama and Saburo Suzuki, /. Antibiotics (Japan) 

8A39 (1955). 

1168 Griseoflavin, colorless needles, m.p. 210-215° (dec). 

Not precipitated by peptide reagents. Negative carbo- 
hydrate and amino sugar tests, FeClg. 
Streptomyces griseoflavus 
Yoshio Waga, /. Antibiotics (Japan) 6A 66 (1953). 

1169 Griseoviridin, C22H29O7N3S (proposed), colorless crystals, m.p. 

(polymorphic') 158-166°, 194-200°, 230°, 240° (dec), 
[oiW -237° (c 0.5 in methanol). 

Neutral compound. Negative FeClg, Sakaguchi, posi- 
tive Bayer. Gives cystine on acid hydrolysis. Further 
structural features are suggested in the last reference 
below. 

StreptoTTiyces griseus, S. griseoviridus n. sp. 

Quentin R. Bartz, Jean Standiford, James D. Mold, Doris W. 
Johannessen, Albert Ryder, Andrew Maretzki and Theodore H. 
HaskeU, "Antibiotics Annual 1954-1955," Medical Encyclope- 
pedia, Inc., New York, p. 777. 

John Ehrlich, George L. Coffey, Myron W. Fisher, Margaret 
M. Galbraith, Mildred Penner Knudsen, Raymond W. Sarber, 
A. S. Schlingman, Robert M. Smith and Jean K. Weston, ibid., 
p. 790 (1954-1955). 

Lucia E. Anderson, John Ehrlich, Sung Huang Sun and 
Paul R. Burkholder, Antibiotics and Chemotherapy 6 100 
(1956). 



Pfizer Handbook of Microbial Metabolites 590 

D. E. Ames, R. E. Bowman, J. F. Cavalla and D. D. Evans, 
J. Chem. Soc, 4260 (1955). 

D. E. Ames and R. E. Bowman, ibid., 4264 (1955). 

1170 Helenine. 

An unstable, little characterized antiviral agent. A 
ribonucleoprotein. 

Penicillium funiculosum 

Richard E. Shope, J. Exp. Med. 97 601, 639 (1953). 

U. J. Lewis, Edward L. Rickes, Laurella McClelland and 
Norman G. Brick, J. Am. Chem. Soc. 81 4115 (1959). 

1171 Heliomycin, needles, chars >300°, complex U.V., Mol. Wt. 235. 

Positive FeClg and Millon. 
May be a polypeptide. 

Actinomyces flavochromo genes var. heliomycini 
M. G. Brazhnikova, T. A. Uspenskaya, L. B. Sokolova, T. P. 

Preobrazhenskaya, G. F. Gauze, R. S. Ukholina, V. A. Shorin, 

O. K. Rossolimo and T. P. Vertogradova, Antibiotiki 3 29 

(1958). 

1172 Hirsutic Acid C, Ci-,H.o04 (proposed), colorless crystals, m.p. 

179.5°, [cz]d'" +li.9° (in absolute ethanol). 

A group of acidic materials. Hirsutic acid C has been 
best characterized. It is a monobasic acid, only slightly 
soluble in H^-O, soluble in most organic solvents. Neg- 
ative 2,4-DNPH, FeCl;^, Fehling. White precipitate with 
Br water. 

Stereum hirsutum 

N. G. Heatley, M. A. Jennings and H. W. Florey, Brit. J. 
Exp. Path. 28 35 (1947). 

1173 Hygroscopin A, C13H.4O3N., oil, b.p. 64° (0.003 mm.), ni.^^ 

1.4830, [a],," 84.7° (in methanol). 

1174 Hygroscopin B, Ci.-.Ho.O^N,, oil, b.p. 70° (0.008 mm.), n„^^ 

1.4935, [a],/' -38.8° (in ethanol). 

Streptoniyces hygroscopicus 

Koichi Nakazawa, Kinzo Oki, Isao Tadokoro, Mikio Honjo, 
Hiroshi Hitomi and Jisaburo Ueyanagi, J. Agr. Chem. Soc. 
Japan 28 296 (1954). 

Sueo Tatsuoka, Akira Miyake, Mikio Honjo, Hiroshi Hitomi, 
Jisaburo Ueyanagi. Masuo Miyamoto, Koiti Nakazawa and 
Kinzo Oki, J. Antibiotics (Japan) 7B 329 (1954). 



591 Unclassified Metabolites 

1175 Hygrostatin, light yellow powder, m.p. 129-131° (dec), [a]ii"" 

+43° (c 1.21 in methanol). 

Contains nitrogen, but no sulfur or halogen. U.V. at 
240 m^. 

Streptomyces Jiygrostaticus 

Kenzo Furushiro, Kiyotake Shimizu, Heiichi Sakai, Masayuki 
Minoyata and Toshio Fujisawa, lyaku, Shigen Kankyusho 
Nevipo 24-39 (1958). (Chem. Abstr. 54 10048b) 

1176 Ilhidin M, Cj,H^.o07, prismatic rods in ethanol, m.p. 216° (cor.)- 

[a],,-" -126° in ethanol. Mol. Wt. 386. U.V. 247, 330 m^ 
in 95 ^f ethanol. 

Contains two acidic groups and an a,/8-unsaturated 
carbonyl group. 

Yield 0.08 g. per liter. 

1177 Illudin S, Ci.-,R.o04, crystalline, m.p. 124-125°. Mol. Wt. 264. 

U.V. 235, 328 m^ in 95 ^c ethanol. 

Yield 0.33 g. per liter. 

Clitocybe illudens 

Marjorie Anchel, Annette Hervey and William J. Robbins, 
Proc. Nat. Acad. Sci. U. S. 36 300 (1950); 38 927 (1952). 

A third, antibiotically inactive substance, Ci,|Hi(.04 or 
CinHo^Oe, crystals, m.p. 72-74°, [aln'" -107° (in absolute 
ethanol) occurred in the same culture. 

1178 Imoticidin, m.p. 245° (darkening from 210°). 

An antibiotic isolate, C 64.71, H 9.50, N 0.0, H.O 7.63. 
Mol. Wt. 532-553. 

Streptomyces albus 

Tadao Inouye, Yasuhiro Okamoto and Yosikazu Nishikado, 
Ber. Ohara Inst. Landjuirtsch. Biol., Okayama Univ. 11 95 
(1959). (In English) 

1179 Indigoidine, deep blue pigment, no melting point. 

Low solubility in most solvents. Soluble in dilute hy- 
drochloric acid. Analysis of partially purified compound: 
C 47.74, H 3.82, N 17.95. Formed a red crystalline ace- 
tate, m.p. >300° (dec), but more soluble: C 49.63, 
H 3.98, N 16.05, acetyl 16.9. A red benzoate was also 
prepared. 

Corynebacterium insidiosiim (McCulloch) Jensen, 
Pseudomonas indigofera, Erwinia chrysanthemi, Arthro- 
bacter sp. 



Pfizer Handbook of Microbial Metabolites 592 

B. Elazari-Volcani, Arch. Mikrobiol. 10 343 (1939). 

D. A. Kuhn and M. P. Starr, Bacteriol. Proc. 58 (1956). 

Mortimer P. Starr, Arch. Mikrobiol. 30 325 (1958). 

1180 Isorhodomycin A,* CooHogOgN or CsiHgiOgN, hydrochloride: deep 

red prisms, m.p. 220°, [aleoeo-Teoo'" +268° ±30° (c 1 in 
methanol). 

Occurs wdth rhodomycin A. 

Either compound on mild hydrolysis yields a water- 
soluble, N-containing moiety and a water-insoluble chro- 
mophore. 

Streptomyces purpurascens 

Hans Brockmann and Peter Patt, Chem. Ber. 88 1455 
(1955). 

1181 Itaconitin, yellow needles, m.p. 169°. 

Negative Beilstein, fuchsin, xanthogen. Legal, Ehrlich, 
Liebermann, FeClg tests. Decolorized bromine and 
KMn04. Formed an acetate, semicarbazone and 2,4- 
DNPH. Hydrogenated to hexahydroitaconitin. 

Aspergillus itaconicus 

Kono Kinoshita and Shoichi Nakajima, Hoshi Yakka 
Daigaku Kiyo 7 17 (1958). 

1 1 82 Laterosporin 

Appeared to be a peptide. Isolated as a hydrochloride. 
Soluble in water. Tendency to precipitate out of solution 
in NaCl solution or in 0.2 m phosphate buffer. 

Bacillus laterosporus 

Ella M. Barnes, "Antibiotics," Vol. II Antibiotics from 
bacteria in the genus bacillus, Oxford University Press, Lon- 
don, 1949, Chap. 10 appendix, pp. 1540-1541. 

1183 Latumcidin (Sulfate), C11H13O2N -112804, white needles, m.p. 

140°, [ocId"' +148.9° (c 0.1 in 0.1 N sodium hydroxide). 

A basic, unstable, antifungal agent. Positive diazo, 
Baeyer, bromine. Negative FeCl^, Fehling, Tollens, Ehr- 
lich, Sakaguchi, ninhydrin, biuret, Molisch. 

Somewhat resembles eulicin, and abikoviromycin. 

Streptomyces reticuli var. latumcidus 

Yoshio Sakagami, Ichiro Yamaguchi, Hiroshi Yonehara, 
Zoichiro Okimoto, Sadazi Yamanouchi, Kazuo Takiguchi and 
Heiichi Sakai, /. Antibiotics (Japan) llA 6 (1958). 
* Identical with entry 597. 



593 



Unclassified Metabolites 



1184 Lenamycin, C4H^0;^No or C4H40._.No (proposed), colorless crys- 

tals, m.p. 202-207° (dec.) optically inactive. 

Apparently an a,/?-unsaturated amide. Negative nin- 
hydrin, biuret, anthrone, FeCl^, Sakaguchi, Elson-Morgan, 
nitro and oxime tests. 

A streptomycete 

The yield was 72 mg. from 5 1. of broth. Occurs to- 
gether wdth fra ns-cinnamic acid amide and ethoxyethene- 
1 ,2-dicarboamide. 

Yasuharu Sekizawa, /. Biochem. (Japan) 45 159 (1958). 

1185 Lenzitin, colorless needles, m.p. 166°. 

Contains C, H, O only. Positive FeClg, KMn04. 

Lenzites sepiaria (Wulf) 

M. Litvinov and E. Moiseeva, Priroda 1 60 (1951). 

1186 Litmocidin, m.p. 144-146° (dec). 

An acid-base indicator. Decolorized by bisulfite or zinc 
dust, color restored by air oxidation. 

Proactinomyces cyaneus var. antibioticiis 

G. F. Cause, /. Bacteriol. 51 649 (1946). 

M. G. Brazhnikova, ibid. 51 655 (1946). (Isolation) 

1187 Longisporin, CgeHsgOio, crystals, m.p. 99-101°, [ajo -f2.62°. 

Alkaline hydrolysis yields a hydroxy acid C^oHi60(OH) 
(COOH). It was suggested that the antibiotic is a cychc 
ester of three such acid units. 

Actinomyces longispori 

G. P. Menshikov and M. M. Rubinshtein, Zhur. Obshchei 
Khim. 26 2035 (1956). 

1188 Lustericin, C4oHti40i:i, white crystals, m.p. 130° [aJD-*" 0°, mol. 

wt. 130. 

Streptomyces sp. 

Motoo Shibata, Koichi Nakazawa, Michitaka Inoue, Jiro 
Terumichi and Akira Miyake, Ann. Rept. Takeda Research Lab. 
17 19 (1958). 

1189 Lycopersin, C^.^HisOg, bright red needles, darkens from 250°, 

dec. 305°. 

Fusarinm lycopersici, F. vasinfectum 

G. Kreitman and F. F. Nord, Arch. Biochem. 21 457 (1949). 

Gerald Kreitman, Oldrlch K. Sebek and F. F. Nord, ibid. 28 
77 (1950). 



Pfizer Handbook of Microbial Metabolites 594 

1190 Malucidin, complex yeast protein, soluble in water, not coagula- 

ble, not dialyzable. Contains organic phosphorus to 
which its activity can be related. 

The protein is combined with RNA, while the latter by 
itself has very little, if any, antibacterial property. 

Brewers' and bakers' yeasts 

I. A. Parfentjev, Federation Proc. 16 428 (1957). 

1191 Marasmic Acid, C10H00O4 (proposed) colorless needles, m.p. 

174° (sealed tube), [x]r>^^ 176° (c 1.4 in acetone). 

A monobasic acid with reducing properties. Negative 
FeCl^, Br^ in CCI4. Forms a 2,4-dinitrophenylhydrazone. 

Marasmius conigenus 

Frederick Kavanagh, Annette Hervey and William J. Rob- 
bins, Proc. Nat. Acad. Sci. U. S. 35 343 (1949). 

1192 Marcomycin, Ci-Ha^OyN:,, white crystals, m.p. 160-180° (dec). 

Streptomyces hygroscopicus 
German Patent 1,027,846 (1958). 

1193 Megacidin, C24H38OJ0 (proposed), colorless crystals, m.p. 162- 

164°, [ain -51° (c 0.958 in ethanol). 

A neutral compound with an easily saponifiable ester 
or lactone group. 

Also isolated from the same fermentation were: L-leu- 

1194 cyl-L-proline anhydride, m.p. 158-165°, [aln —128° (c 

1195 0.968 in ethanol) and L-leucyl-L-leucine anhydride. 

Streptomyces sp. 

L. Ettlinger, E. Gaumann, R. Hiitter, W. Keller-Schierlein, 
F. Kradolfer, L. Neipp, V. Prelog, P. Reusser and H. Zahner, 
Monatsh. Chem. 88 989 (1957). 

1196 Melanosporin, C56-63H105-117O20-22N3, yellowish white amorphous 

solid, m.p. 132-134°, [aW° +30° (c. 1.57 in methanol). 

Strong" acid hydrolysis yields three ninhydrin-positive 
compounds. Negative FeClg. Positive ninhydrin. 

Streptomyces melanosporus {sine m,elanosporofaciens) 
n. sp. 

F. M. Arcamone, C. Bertazzoli, M. Ghione and T. Scotti, 
Giorn. microbial. 7 207 (1959). 

1197 Mesenterin, colorless needles, m.p. 122-126°. 

A basic compound, analysis: C 65.82, H 7.10, N 8.66. 
Positive Molisch, negative ninhydrin, biuret, Fehling, 
Feci,. 



595 



Unclassified Metabolites 



Occurs with azomycin and antibiotic 446. 
Nocardia mesenterica 

Masahiro Ueda and Hamao Umezawa, /. Antibiotics (Ja- 
pan) 8A 164 (1955). 

1198 Metabolite, C24H-0O2, colorless crystals, m.p. 82°. 

Negative Liebermann-Burchard, KMnO,, tetranitro- 
methane tests. 

Amanita phalloides 

Heinrich Wieland and Gustav Coutelle, Ann. 548 270 ( 1941 ). 

1199 Metabolite of Coprinns comatis, Ci2Hi,.0N<>S, m.p. 157°. 

A basic compound, containing a phenolic hydroxyl 
group. Positive Millon, Pauly diazo tests. Raney nickel 
desulfurization gave a compound, m.p. 250° (dec). 

Copriniis comatis Gray 

Paul Heinz List, Arch. Pharm. 291 502 (1958). 

1200 Metabolite from Curvularia lunata, Ci4His,05, colorless needles, 

m.p. 195°. 

Insoluble in aqueous sodium carbonate, soluble in aque- 
ous sodium hydroxide. Brown color with alcoholic ferric 
chloride. 

Curvularia lunata 

Also isolated from the same culture were mannitol and 
a trace of crystalline material, m.p. 176-178° (dec). 

T. Krishna Murty and S. Sankara Subramanian, Indian J. 
Pharm. 20 72 (1958). 

1201 Metamycin, white crystals, m.p. 173° (dec.) [ajr^^^ +36.6 (c 

d.ll in methanol) U.V. 237, 305-307 m^ in 0.1 N sodium 
hydroxide. 

C 43.95, H 4.06, N 14.45, S 13.57. 

Positive Fehhng, Tollens, Bro, decolorization of per- 
manganate, 2,4-DNPA tests. Negative FeClg and Saka- 
guchi tests. 

Streptomyces matensis 

P. Sensi, R. Ballotta and G. G. Gallo, Antibiotics and Chemo- 
therapy 9 76 (1959). 

1202 Microcin A, neutral, reddish violet in color, separated at pH 7.0. 

1203 Microcin B, acidic, yellowish red, slightly soluble in water sepa- 

rated at pH 2.0. 

Both give negative Molisch and FeClg; vary from mi- 
cromonosporin in activity, have much resistance to U.V. 



Pfizer Handbook of Microbial Metabolites 596 

Micromonospora sp. 

Tomotsune Taira and Shigehiro Fujii, J. Antibiotics (Japan) 
5 187 (1952). 

1204 Mikamycin A, C31H39O9N3, yellowish white crystals, m.p. 147- 

152° (dec), [aln'" -152° (c 0.5 in methanol). 

Apparently identical with the principal active com- 
ponent of the streptogramin and antibiotic No. 899 com- 
plexes. 

A neutral antibiotic. Negative ninhydrin, biuret, glu- 
cosamine, maltol and Millon. Green-black FeCla. Brown 
precipitate with the Tollens reagent. Positive Benedict. 
Forms a 2,4-DNPH. 

Streptomyces mitakaensis 

Mamoru Aral, Keiko Karasawa, Shoshiro Nakamura, Hiroshi 
Yonehara and Hamao Umezawa, J. Antibiotics (Japan) 11 A 
14 (1958). 

Mamoru Aral, Koichi Okabe, Hiroshi Yonehara and Hamao 
Umezawa, ibid. IIA 21 (1958). 

Koichi Okabe, ibid. 12A 86 (1959). 

1205 Mikamycin B, C45H5sOiiNg (proposed), white platelets, m.p. 

160°, dec. 262°, [aW -61.3° (c 1.0 in methanol). 

Similar to PA-114 B in physical and chemical proper- 
ties but differs from staphylomycin S. It is thought to be 
different from both. 

Gives a positive FeClj. Negative Ehrlich, biuret, 
Fehling, Tollens, nearly negative ninhydrin. 

Streptomyces mitakaensis 

Kiyoshi Watanabe, Hiroshi Yonehara, Nobuo Tanaka and 
Hamao Umezawa, J. Antibiotics (Japan) 12A 112 (1959). 

Koyoshi Watanabe, ibid. 13A 57 (1960). 

Mitomycins. 

1206 A complex from which several compounds were iso- 

1207 lated: colorless fractions W-1 (m.p. 148°), W-2 (m.p. 

1208 138°) and W-3 (m.p. 187°). Pigmented fractions A (red 

1209 crystals) m.p. 167°, B (violet crystals), C (bluish violet 

1210 crystals), Y (yellow crystals) m.p. 180-240° (dec.) and 

1211 R (red-brown amorphous powder). Pigmented fractions 

1212 are antibiotic. 

1213 

1214 Mitomycin C, C54H61OJ9N13 (tentative), deep bluish violet crys- 
tals, m.p.: no melting or dec. noted below 360°. 



597 Unclassified Metabolites 

:• Positive FeCl;,, Fehling, biuret, Ehrlich, decolorization 

of permanganate. Negative Benedict, Tollens, ninhydrin, 
Milton, Raymond. Mol. Wt. ~1120. 

The mitomycins may be related to the actinomycins. 

Streptomyces caespitosus 

S. Wakaki, H. Marumo, K. Tomioka, G. Shimizu, E. Kato, 
H. Kamada, S. Kudo and Y. Fugimoto, Antibiotics and Chemo- 
therapy 8 228 (1958). 

Toju Hata, Yoshimoto Sano, Ryozo Sugawara, Akihiro 
Matsumae, Kokichi Kamamori, Tatsuo Shima and Tadashi 
Hoshi, ;. Antibiotics (Japan) 9A 141 (1956). 

1215 INIoldin, gives positive Molisch and FeCls but negative biuret, 

ninhydrin, Tollens, Fehling and Sakaguchi tests. 
Streptomyces sp. res. S. phalochroviogenus 
Kenji Maeda, Yoshiro Okami, Osamu Taya and Hamao 

Umezawa, J. Antibiotics (Japan) 5 465 (1952). 

1216 Monilin, Ci^HooOgNfi, colorless needles, m.p. 235-238° (dec). 

An antifungal compound. Positive ninhydrin. 

Streptomyces sakaiensis 

Shigehiro Fujii, Hiromu Hitomi, Masahiko Imanishi and 
Koichi Kakazawa, Ann. Rept. Tdkeda Research Lab. 14 8 
(1955). 

1217 Musarin (C35H6oOi4N2)n (proposed), Mol. Wt. ^5000, yellow 

powder, m.p. 170° (dec), UW" +35.1° ±1.6° (c 1.21 in 
methanol). 

An acidic substance. 

Streptomyces sp. 

H. R. V. Arnstein, A. H. Cook and Margaret S. Lacey, J. Gen. 
Microbiol. 2 111 (1948). 

1218 Mutomycin, C7H11.12O2, white crystalline powder, m.p. 141.5- 

142°. 

Actinomyces atroolivaceus var. mutom.ycini 

G. F. Gauze, T. S. Maksimova, O. L. Popova, M. G. Brazh- 

nikova, T. A. Uspenskaya and O. K. Rossolimo, Antibiotiki 

U.S.S.R. 4 20 (273 in EngUsh) (1959). 

1219 Mycelin, m.p. 263° (dec). 

Water insoluble, contains no nitrogen or sulfur. Neg- 
ative Molisch. Flavomycin is produced by the same or- 
ganism. Mycelin has antifungal properties. 

Streptom.yces roseofiavus 

Kazuyoshi Also, Tadashi Aral, Kazuhiro Washlda and Tei 
Tanaami, J. Antibiotics (Japan) 5 217 (1952). 



Pfizer Handbook of Microbial Metabolites 598 

1220 Mycelin-IMO, yellow crystalline, m.p. 214-222° (dec), [a]n^^ 

+70 ±2 (c 1 in 1,4-dioxane) U.V. 243, 294, 335, 355, 373 
m/j,. Mol. Wt. 335, C 71.29, H 5.96, N 11.31. 

Streptomyces diastatochromogenes 

Koichi Ogata, Masaji Igarashi, Akira Miyake and Hiroichi 
Yamamoto, Japanese Patent 5898 (1957). 

1221 Mycorhodin, bright red needles, m.p. 200-202° (dec.) U.V. 

420, 471, 250m;x in ethanol. C 58.7 H 5.2 N 2.1. 

Mol. Wt. 698, 635. 

Acid-base indicator. 

Streptomyces sp. 

M. Misiek, A. Gourevitch, B. Heinemann, M. J. Cron, D. F. 
Whitehead, H. Schmitz, I. R. Hooper and J. Leln, Antibiotics 
and Chemotherapy 9 280 (1959). 

1222 Mycospocidin (C20H32O9N2),,, colorless crystals, dec. 233°, 

[a]D-" +56° "(c i in pyridine). 

Negative ninhydrin, biuret, Tollens, Fehling, ferric 
chloride tests. Positive diazo reaction. 

Acid hydrolysis yielded two ninhydrin-positive sub- 
stances, one perhaps being glycine. 

Streptomyces bobiliae 

Shoshiro Nakamura, Mamoru Aral, Keiko Karasawa and 
Hiroshi Yonehara, J. Antibiotics (Japan) lOA 248 (1957). 

1223 Mycothricin, colorless crystals, complex consists of strong or- 

ganic bases. 

Negative ninhhydrin, biuret, Fehling, Tollens, Molisch, 
Millon, maltol and Sakaguchi. 

Streptomyces lavendulae 

G. Rangaswami, Hindustan Antibiotics Bull. 2 46 (1959). 

1224 Mycoticin, CigHsoO-, (proposed), yellow crystals. 

Contains a hydroxyl group, has reducing properties, 
fluoresces under U.V. 

Streptomyces ruber 

Ruth C. Burke, Jacob H. Swartz, S. S. Chapman and Wei- 
Yuan Huang, J. Invest. Dermatol. 23 163 (1954). 

1225 Nigericin, C3i,H,;,,0i,, colorless needles, m.p. 246-254°. 

A monobasic acid. 

Streptomyces sp. resembling S. violaceaniger 
Roger L. Harned, Phil Harter Hidy, Cyril J. Corum and Ken- 
neth L. Jones, Antibiotics and Chemotherapy 1 594 (1951). 



599 Unclassified Metabolites 

1226 Nocardianin, C6r,.67H»6.i04Oi5Ni8, red prisms, m.p. 228-235° 

(dec), [a]i."" -223° (c 0.3 in methanol). 

Negative biuret, ninhydrin. 

Nocardia sp. 

I. R. Bick, Gregory J. Jann and Donald J. Cram, Antibiotics 
and Chemotherapy 2 255 (1952). 

1227 Nocardorubin, crimson powder, darkens from 180° (dec.). 

Nocardia narasinoerisis 

J. Antibiotics (Japan) 8B 253 (1955). 

1228 Nonactin, C;,oH^^O,,, colorless crystals, m.p. 147°, optically in- 

active. 

Slight U.V. at 264 m^ (log e = 1.5 in ethanol). Inert 
to chemicals and microbes. 

Streptomyces spp. which produce cycloheximide. 

R. Corbaz, L. Ettlinger, E. Gaumann, W. Keller-Schierlein, 
F. Kradolfer, L. Neipp, V. Prelog and H. Zahner, Helv. Chini. 
Acta 38 1445 (1955). 

1229 Nordin, CigHieO^sCL, m.p. 134-136°. 

Occurs with estin (q.v.). 

Penicilliuin paxilli var. echinulatum 

Eitaro Komatsu, Japanese Patent 4799 (1953). 

1230 Nudic Acid A, C,4H2i,0;^ (proposed), colorless crystals, m.p. 

123.5°. 

No reducing properties. Takes up bromine. 

Tricholoma nudum 

H. W. Florey, E. Chain, N. G. Heatley, M. A. Jennings, A. G. 
Sanders, E. P. Abraham and M. E. Florey, "Antibiotics," Oxford 
University Press, London, 1949, p. 358. 

1231 Nybomycin, C1SH14O4N0, colorless crystals, which darken at 330° 

without melting. 

Negative ninhydrin, biuret, FeClg; sugar tests, Ehrlich, 
KM,.04, Bro. 

Streptomyces sp. 

Frieda Strelitz, Helen Flon and Igor N. Asheshov, Prac. Nat. 
Acad. Sci. U. S. 41 620 (1955). 

T. E. Eble, G. A. Boyack, C. M. Large and W. H. De Vries, 
Antibiotics and Chemotherapy 8 627 (1958). 

1232 Oligomycin A, C24H4yO(., colorless crystals, m.p. 140° (dec), 

150° (dec.) (polymorphic), [alo"^ —54.5° (c 4.40 in diox- 
ane). 



Pfizer Handbook of Microbial Metabolites 600 

Mol. Wt. = 424. Absorbs 2 moles H2. Four active H. 
Five C — CH3 groups. Forms a diacetate. 

1233 Oligomycin B, C22H36O6, colorless crystals, m.p. 160°, 169° 

(polymorphic), [aW^^ —49.5° (c 1.03 in methanol). 
Mol. Wt. = 396. Four active H. Five C— CH3 groups. 
Forms a diacetate. 

1234 Oligomycin C, CorH^.-Oc colorless crystals, m.p. 198-200°, [aW^ 

-80.7° (c 3.70 in dioxane). 

Contains six C — CH3 groups. 

Streptomyces sp. (may be S. diastatochromo genes) 

Robert M. Smith, William H. Peterson and Elizabeth McCoy, 
Antibiotics and Chemotherapy 4 962 (1954). 

Satoru Masamune, J. M. Sehgal, E. E. van Tamelen, F. M. 
Strong and W. H. Peterson, /. Am. Chem. Sac. 80 6092 (1958). 

1235 Ophiobalin, C28H30O4, white prisms, m.p. 181-182°. 

Ophiobalus miyabeanus 

A. Neelameghan, Hindustan Antibiotics 2 13 (1959). 

1236 Oregonensin, C20H32OS (proposed), colorless needles, m.p. 82°. 

A neutral substance. Positive 2,4-DNPH. 

Ganoderma oregonense 

H. W. Florey, E. Chain, N. G. Heatley, M. A. Jennings, A. G. 
Sanders, E. P. Abraham and M. E. Florey, "Antibiotics," Oxford 
University Press, London, 1949, p. 362. 

1237 Oryzacidin (Oryzasizine), CgHigOsN, colorless, hygroscopic nee- 

dles, m.p. 162° (dec), [ajn -138°. 

y3-Nitropropionic acid also occurs free in the culture 
broth. 

Aspergillus oryzae 

Chujiro Shimoda, /. Agr. Chem. Soc. Japan 25 254 (1951). 

Seiji Nakamura and Chuji Shimoda, ibid. 28 909 (1954). 

1238 PA-128, C37.46H6i.75O13.1cN, Ught yellow rectangular plates, m.p. 

143° [aln'' -2.0° (c 1 in methanol). 

Negative FeClg, no colors in aqueous base nor concen- 
trated H2SO4. Positive 2,4-DNPH, decolorizes Brs water 
and permanganate. Takes up >6 mM of hydrogen per 
gram of antibiotic. 

Unclassified Streptomycete 

Koppaka V. Rao and John E. Lynch, Antibiotics and Chemo- 
therapy 8 437 (1958). 



6oi Unclassified Metabolites 



i239 PA-132, CicHissoOr,, free acid is a colorless amorphous powder, 
[a].,-' —161° (c 1.0 in methanol). Handled as the ben- 
zylamine salt: white crystals, m.p. 128-131°, [a]i)~^ —130° 
(c 1.0 in methanol). 

A lactonic acid containing two C-methyl groups. De- 
colorizes bromine or permanganate. Negative FeCla, 
Fehhng, 2,4-DNPH, Tollens, AgNOg and NaOI. 

Streptomyces sp. 

B. Kenneth Koe, Ben A. Sobin and Walter D. Celmer, "Anti- 
biotics Annual 1956-1957," Medical Encyclopedia, Inc., New 
York, p. 672. 

1240 Phagolessin A 58, light yellow hygroscopic powder. 

Negative FeClg, biuret, Millon and ninhydrin test. 
Streptomyces sp. 

Igor N. Asheshov, Freda Strelitz and Elizabeth A. Hall, Anti- 
biotics and Chemotherapy 2 366 (1952). 

1241 Phalamycin, CaeH^iOi^N^S (proposed), colorless crystals, no 

sharp m.p. 

Positive FeClg, Bro absorption. Has primary or second- 
ary alcohol groups. 

Streptomyces noursei variant 

Rachel Brown, N. Y. State Dept. Health, Ann. Kept. Div. 
Labs and Research 18 (1956). (Chem. Abstr. 51 16672e) 

1242 Phalofacin gives positive FeCls but negative biuret, Millon, nin- 

hydrin, MoHsch, Tollens and Sakaguchi tests. 

Streptomyces sp. res. S. aureus 

Kenji Maeda, Yoshiro Okami, Osamu Taya and Hamao 
Umezawa, /. Antibiotics (Japan) 5 465 (1952). 

1243 Phleomycin, C53H93O32N17, white to pale green amorphous pow- 

der, isolated as a blue monocopper complex. U.V. 244, 
295-300 lUfi. 

Gives positive ninhydrin and diazo tests. Negative 
Fehling, Tollens, Sakaguchi and Molisch. 

Streptomyces verticillis 

Tomohisa Takita, Kenji Maeda and Hamao Umezawa, J. 
Antibiotics (Japan) 12A 111 (1959). 

Tomohisa Takita, ibid. 12 285 (1959). 

1244 Phytonivein, C29H46O2, colorless needles, m.p. 138°. 

Fusarium bulbigenum 
The watermelon wilt toxin. 



Pfizer Handbook of Microbial Metabolites 602 

Isamu Hirose and Seiyo Aoe, Ann. Phytopathol. Soc. Japan 
19 162 (1955). (Chem.' Abstr. 50 14058g) 

Isamu Hirose and S. Nishimura, Nippon Nogii-kagaku 
Kaishi 30 528 (1956). 

1245 Piricularin, C17H14N2O3 or C18H14N0O3, colorless crystals, m.p. 

73.5°, [a]D'« -19°. 

Absorbs 4 moles of hydrogen over platinum catalyst, 
contains two phenolic or enolic hydroxyls, no methoxyl. 
Reacts with 3 moles of 2,4-dinitrophenylhydrazine. Has 
1 N-methyl, no NH or NHo. A max. in HoO = 240 m/x. 
Ejom/"^^ 2824. A toxin of rice blast disease. 

Piricularia oryzae 

Kinjiro Tamari and Jun Kaji, Nippon Nogei-kagaku Kaishi 
31 387 (1957). 

1246 neomycin, C,4Hj20s, rectangular plates from ethanol, m.p. 

235°, U.V. 270, 330, 340 m^^ in 0.13 m phosphate buffer. 

Streptomyces pleofaciens 

Roy A. Machlowitz, Jesse Charney, Alfred A. Tytell and 
W. P. Fisher, "Antibiotics Annual 1954-1955," Medical Ency- 
clopedia, Inc., New York, p. 806. 

1247 Pleuromutilin (Drosophilin B), C22H340r,, colorless crystals, 

m.p. 170°, [aW +20° (c 3.0 in absolute ethanol). 

Forms a diacetate, non-phenolic, probably has a lactone 
ring, forms a hydrazone. 

Pleurotus mutilus 

Marjorie Anchel, /. Biol. Chem. 199 133 (1952). 

1248 Pleurotin, C20H22O5, yellow-amber needles, m.p. 220-215° 

(dec), [a]v~^' —20° (c 0.59 in chloroform). 

A neutral, photosensitive compound. Negative FeCls, 
oxidized KI. 

Pleurotus griseus 

William J. Robbins, Frederick Kavanaugh and Annette 
Hervey, Proc. Nat. Acad. Sci. U. S. 33 171 (1947). 

1249 Pluramycin A, orange needle crystals, dec. from 177°, U.V. 208, 

245 (265-270) m^^ in ethanol. 
C 66.63, H 6.30, N 3.66 
Negative FeCl^, Fehling, ToUens and 2,4-DNPH. 

1250 Pluramycin B, reddish brown powder, possible neutral sub- 

stance. The pluramycins may be related to the actino- 
mycins. 



6o3 Unclassified Metabalites 

Tomio Takeuchi, Kazuo Nitta and Hamao Umezawa, J. Anti- 
biotics (Japan) 9 A 22 (1956). 

Kenji Maeda. Tomio Takeuchi, Kazuo Nitta, Koki Yagishita, 
Ryozo Utahara, Teisuke Osato, Masahiro Ueda, Shinichi Kondo, 
Yoshiro Okami and Hamao Umezawa, ibid. 9A 75 (1956). 

1251 Poin, crystals, m.p. 142-143°. 

C 59.70, H 7.77, O 32.53 
Fusarium sporotrichiella var. poae 

O. K. filpidina, Antibiotiki U.S.S.R. 4 46 (273 in English) 
(1959). 

1252 Primycin, C,.,H.5-0;N, white microcrystals, m.p. 166-168° (dec). 

No reducing properties. Can be acetylated. Strong 
Sakaguchi test. 

An unclassified actinomycete 

T. Valyi-Nagy, J. Ori and I. Szilagy, Nature 174 1105 (1954). 

1253 Psalliotin, crystalline, water soluble, inactivated by bright light. 

Psalliota xanthoderma 

Nancy Atkinson, Nature 174 598 (1954). 

Idem., Australian Patent 20,272,156 (1957). 

1254 Pulvilloric Acid, buff colored needles, turning bright yellow in 

air. 

An acidic, antifungal antibiotic, containing only C, H, 
O. Blue FeCli, negative Tollens. Yield 600 mg. per liter. 

PenicilliuTn pulvillorum Turfitt 

P. W. Brian, P. J. Curtis, H. G. Hemming and G. L. F. Norris, 
Brit. Mycol. Soc. Trans. 40 369 (1957). 

1255 Pumilin, lemon-yellow crystals, m.p. >360°. 

Negative FeCl-,, copper-red in 5 N hydrochloric acid. 

Bacillus pumilis 

0.7 g. was obtained from 500 gal. of broth. 

D. S. Bhate, Nature 175 816 (1955). 

1256 Racemomycin A 

1257 Racemomycin B,* C60H128O32N20, white powder, m.p. (Hydro- 

chloride) 175° (dec), [o^W -45° (c 0.5 in water). 

Positive Molisch, Elson-Morgan and biuret. Negative 
Sakaguchi, maltol, FeCl^, 2,4 DNPH and Fehhng. Yields 
^-lysine and roseonine on hydrolysis. 

1258 Racemomycin C, isolated in a small amount as a salt (m.p. 

210°). 
* See entry 790. 



Pfizer Handbook of Microbial Metabolites 604 

Streptomyces racemochromogenes n. sp. 
Hyozo Taniyama and Shoji Takemura, J. Pharm. Soc. Japan 
77 1210, 1217 (1957); 78 742 (1958). 

1259 Ractinomycin A, C33H3nOi4N3, orange needles, m.p. browns 

-157°, blackens at 205°. 

Negative ninhydrin, biuret, Sakaguchi, Millon. Posi- 
tive Tollens, Molisch, FeClg. Decolorizes KMn04. De- 
colorized by HoOo. Alkali-unstable. Turns purple above 
pH 6.5. Contains no amino acids. 

1260 Ractinomycin B, reddish orange needles, m.p. 172-175° (dec). 

Negative FeClg. 

The ractinomycins are said to resemble the actinomy- 
cins in some respects. 

Streptomyces sp. similar to S. phaeochromogenes 

Ryozo Utahara, Hideo Oyagi, Koki Yagishita, Yoshiro Okami 
and Hamao Umezawa, J. Antibiotics (Japan) 8A 132 (1955). 

Ryozo Utahara, ibid. lOA 115 (1957). 

S. Wakiki et al., Antibiotics and Chemotherapy 8 228 
(1958). 

1261 Raisnomycin, dark yellow basic material, insoluble in water. 

The hydrochloride and disulfate are slightly soluble. The 
impure material does not have an end absorption in U.V. 

Streptomyces kentuckensis 

Fred S. Barr and Paul E. Carman, Antibiotics and Chemo- 
therapy 6 286 (1956). 

1262 Rammacin, C26H43O8, crystalline, m.p. 235°, Mol. Wt. 499. 

Negative Brs; positive benzenoid. 

Streptomyces sp. 

K. Ahmad and M. F. Islam, Nature 176 646 (1955). 

1263 Ramycin (Mol. Wt. 478, contains only carbon, hydrogen and 

oxygen), 'colorless plates, m.p. 158° (dec), optically in- 
active. 

Structural features: 

A non-phenolic hydroxy acid with one or more carbon- 
carbon double bonds. 

Mucor ramannianus 

P. J. van Dijck and P. deSomer, J. Gen. Microbiol. 18 377 
(1958). 

1264 Raromycin, m.p. 211-213° C 57.97, H 8.46, N 0.44, O 33.13 by 

difference. 



6o5 Unclassified Metabolites 

'.' Streptomyces sp. 

Nabuo Tanaka, Hisaji Yamazaki, Koichi Okabe and Hamao 
Umezawa, /. Ajitibiotics (Japan) lOA 189 (1957). 

1265 Roseomycin, crystalline helianthate, m.p. 211-216° (dec.) and 

reineckate m.p. 114° (dec). 

Positive Molisch, Tollens, indole, glucosamine and 
Fehling. 

Negative maltol, biuret, ninhydrin and Sakaguchi. 

Streptomyces roseochromogenes 

Nakao Ishida, J. Antibiotics (Japan) 3 845 (1950). 

1266 Rhizobacidin, crystalline, m.p. 215-220° (dec). Contains C, 

H, O and N but not S. Positive biuret, xanthoproteic, 
ninhydrin and Sakaguchi. Negative Ehrlich, Molisch and 
FeCla. 

Bacillus subtilis 

Carlos Casas-CampiUo, Ciencia (Mexico) 11 21 (1951). 

1267 Rhodocidin, red powder, U.V. shows a broad peak at 500-530 

mix. Soluble in water and organic solvents. 

Streptomyces phoenix 

Jesse Charney, Roy A. Machlowitz, W. S. Roberts and W. P. 
Fisher, Antibiotics and Chemotherapy 3 788 (1953). 

Ristocetins (Spontins, Ristins). 

Two closely related amphoteric antibiotics containing 
amino and phenolic groups and sugars. Each contains 
four reducing sugars: glucose, mannose, rhamnose and 
D-arabinose. 

Negative biuret, Sakaguchi, maltol. Positive phospho- 
molybdic acid test for phenols, ninhydrin (after acid hy- 
drolysis), anthrone. Mol. Wt. 2500-5000. Contain C, H, 
O, N, S. 

1268 Ristocetin A (Sulfate): [alo'^ -120-133° (in water). 

1269 Ristocetin B (Sulfate): [ajn" -144-149° (in water). 

Nocardia lurida 

Julian E. Philip, Jay R. Schenck and Martha P. Hargle, 
"Antibiotics Annual 1956—1957," Medical Encyclopedia, Inc., 
New York, p. 699. 

1270 Rotaventin, white crystals, m.p. 170-175° (dec). 

Streptom.yces reticuli 

Nobukiko Komatsu and Momoe Soeda, Japan. J. Exp. Med. 
21 279 (1951). 



Pfizer Handbook of Microbial Metabolites 606 

1271 Rubromycin, thin square rods, m.p. 215° (dec.) U.V. 518-520, 

546, 584 m^. 

C 60.30, H 4.26, O 33.91 

Contains no N (differing from rhodomycin). Differs 
from rhodomycetin in that the latter is found in the cul- 
ture solution; the present compound is in the mycelium. 

Streptomyces collinus n. sp. 

Hans Brockman and Karl Heinz Renneberg, Naturwissen- 
schaften 40 59 (1953). 

1272 Ruticin, orange needle-hke crystals, U.V. 227, 262, 364 m^x. 

Streptomyces res. S. rutgersensis 

W. P. Fisher, Jesse Charney, Ray A. Machlowitz, James E. 
Blair and Alfred A. Tytell, "Antibiotics Annual 1953-1954," 
Medical Encyclopedia, Inc., New York, p. 174. 

1273 Sarcidin, m.p. 274-275° (dec). 

C 41.89, H 5.02, N 21.82 and a qualitative sulfur test. 
Tamio Takeuchi, Kazuo Nitta and Hamao Umezawa, /. Anti- 
biotics (Japan) 6A 31 (1953). 

1274 Secalonic Acid, C31H30.32O14, lemon-yellow needles, m.p. 244- 

250° from chloroform, [a],,-" -81° (acetone), -66° (chlo- 
roform), -198°->-59° (pyridine). 

Claviceps purpurea 

F. Kraft, Arch. Pharm. 244 336 (1906). 

A. Stoll, J. Renz and A. Brack, Helv. Chim. Acta 35 2022 
(1952). 

1275 Seligocidin, crystalline powder, U.V. 304 m^j, in ethanol. 

Positive Sakaguchi and ninhydrin; negative biuret. 
Streptomyces res. S. roseochromogenes 
Shoshiro Nakamura, Kenji Maeda, Yoshiro Okami and 
Hamao Umezawa, J. Antibiotics (Japan) 7A 57 (1954). 

1276 Sirenin, CoiHsgO^N. 

Mol. V^t. : found 386, calculated 414. Contains a lac- 
tone ring, a carbonyl group and a — C=C — or — C=N — 
bond. The absence of hydroxyl and carboxyl groups and 
of aromatic rings was ascertained. 

Allomyces species 

Sirenin is a sex hormone of this water-mold. 

Leonard Machlis, Nature 181 1790 (1958). 



6o7 Unclassified Metabolites 

1277 Sporidesmin (probably) Ci9H:;iO,iN;<SXlCCl4, colorless crystals 

(carbon tetrachloride solvate) sintering from 109°-^ 
resins 125° semi-solid -^ meniscus at 130-134°, [a]i.-" 
-19° (c 2.2 in methanol). 

Other formulae without chlorine are not excluded, 
since the solvent-free compound has not been isolated. 
The compound is a toxin in animals. 

Sporidesmiiim bakeri Syd. 

R. L. M. Synge and E. P. White, Chem. and Ind., 1546 
(1959). 

1278 Streptocardin, Crystalline, U.V. 365 (242) (252) m^^ in phos- 

phate buffer (pH 6) forms water-soluble alkali salts. 

Streptomyces sp., Nocardia sp. 

W. P. Fisher, Roy A. Machlowitz, Alfred A. Tytell and Jesse 
Charney, "Antibiotics Annual 1953-1954," Medical Encyclope- 
dia, Inc., New York, p. 177. 

1279 Streptolydigin, C;i.H4e09No (or C3,-H.-^oOioNo), m.p. 144-150° 

(dec), [alo"' —93° (c 1.6 in chloroform). 

An enolic acid. Positive FeCls, iodoform. Negative 
biuret, ninhydrin, Fehling, Molisch. Reacts with Bto in 
CCI4. 

Streptomyces lydicus 

T. E. Eble, C. M. Large, W. H. DeVries, G. F. Crum and 
J. W. Shell, "Antibiotics Annual 1955-1956," Medical Encyclo- 
pedia, Inc., New York, p. 893. 

Streptovaricin (Dalacin). A complex consisting of at least five 
active closely related components. These were separated 
by countercurrent distribution into Streptovaricins : 

1280 A, C34H4-.490i;^N, yellow crystals, m.p. 182-184°, [2W +454° 

(CHCI3). 

1281 B, C34H4749O13N, yellow crystals, m.p. 195-200°, [aW* -fl68° 

(CHCI3). 

1282 C, C34H4-.4c,0,3N, yellow crystals, m.p. 168-171°, [ajir" +317° 

(CHCI3). 

1283 D, yellow crystals, m.p. 115-118°, hW^ +102° (CHCI3). 

1284 E, yellow crystals, m.p. 102-105°, [aW +6.13° (CHCI3). 



Pfizer Handbook of Microbial Metabolites 608 

Streptoinyces spectabilis 

Paul Siminoff, Robert M. Smith, Walter T. Sokolski and 
G. M. Savage, Am. Rev. Tuherc. Pulmonary Diseases 75 576 
(1957). 

George B. Whitfield, Edward C. Olson, Ross R. Herr, John A. 
Fox, Malcolm E. Bergy and Gerald A. Boyack, ibid. 75 584 
(1957). 

Upjohn Co., British Patent 811,757 (1959). 

1285 Streptozotacin, C14H17O12N5, m.p. 115-125° (dec). 

Probably still a mixture. Base-unstable neutral 
substance. Seems to contain the partial structure 


R — Cf /N=0. Alkaline treatment liberates diazo- 

^CH3 
methane. 

Streptomyces achromogenes 

R. R. Herr, T. E. Eble, M. E. Bergy and H. K. Jahnke, 7th 
Annual Symposium on Antibiotics, Washington, D. C, 1959. 

1286 Substance 1404, yellow crystalline, Hexaene. M.p. 210-220° 

(dec), [aln'' +67.5 ±2.0° (c 1 in dioxane). 
Contains N 10.47, no sulfur, no halogen. 
Streptomyces diastatochromogenes (Mycelium) 
Masaji Igarashi, Koichi Ogata and Akira Miyake, J. Anti- 
biotics (Japan) 8B 113 (1955). 

1287 Sulfactin, C3sH.-r,07NiiS4 or C27H4oO,-,NsS3 (proposed), hygro- 

scopic white needles, m.p. 245-275° (dec). 

Positive Fehling. Reduces KMn04. Negative biuret, 
FeCl3, Molisch, Sakaguchi. 

Streptomyces roseus 

Renate Junowicz-Kocholaty, Walter Kocholaty and Albert 
Kelner, J. Biol. Chem. 168 765 (1947). 

1288 Sulfocidin, yellow-brown crystals, m.p. 166-178°, [a]D^^ —58.5° 

(c 0.51 in chloroform). 

Neutral antibiotic, analysis C 64.88, H 8.38, N 4.25, S 
1.80. Negative nitroprusside and azide iodine, ninhydrin, 
FeCl3, Sakaguchi, maltol, biuret, Fehling, 2,4-DNPH. De- 
colorizes permanganate. 

Streptomyces sp. 

Morris Zief, Robert Woodside and George E. Ham, "Anti- 
biotics Annual 1957-1958," Medical Encyclopedia, Inc., New 
York, p. 886. 



6o9 Unclassified Metabolites 

1289 Taitomycin, yellow-brown powder, U.V. at 330, 420 m^x. 

C 53.57, H 4.87, N 9.50 ash 2.8. 

Positive Fehling and ninhydrin (acid hydrolysate). 

Streptoynyces afgJianensis 

Mitsuo Shimo, Tatsuji Shiga, Takashi Tomosugi and Ikuzo 
Kamoi, /. Antibiotics (Japan) 12A 1 (1959). 

Takashi Tomosugi, Ikuzo Kamoi, Tatsuji Shiga and Mitsuo 
Shimo, ibid. 12A 7 (1959). 

1290 Tardin, CnHir.Oa (proposed), pale yellow oil, [a]v^° —11.4° (in 

alcohol). 

Positive FeCl... Negative 2,4-DNPH. Hydrolyzes to an 
acidic and a neutral fraction. 

PeniciUium tardum 

N. Borodin, F. J. Philpot and H. W. Florey, Brit. J. Exp. 
Path. 28 31 (1947). 

1291 Terrecin, light yellow prisms, m.p. 219°. 

Analysis: C 51.89, H 3.51, N 3.8, CI 19.1. Alkali 
soluble. Positive FeCl^. 

Aspergillus terrens 

Kazuo Iwata and Itiro Yosioka, /. Antibiotics (Japan) 3 192 
(1950). 

1292 Thiactin, acid and alkali metal salts (previously identified as 

bryamycin). M.p. 220-234°, [a]n'' -68.5 -69.5° (c 1 
in chloroform). 

Streptomyces hawaiiensis 

Bernard Heinemann, Irving R. Hooper and Martin J. Cron, 
British Patent 790,521 (1958). 

1293 Thioaurin (Orosomycin, Antibiotic HA-9), CyHeOoN^So or 

Ci4H,.j04N4S4 (proposed), yellow crystals, m.p. 178-180°, 
optically inactive. 

Strong U.V. at 232, 370 m^. Negative FeClg. 

Streptomyces sp. resembling S. lipmanii 

William A. Bolhofer, Roy A. Machlowitz and Jesse Charney, 
Antibiotics and Chemotherapy 3 382 (1953). 

William Eisenman, P. Paul Minieri, Anthony Abbey, John 
Charlebois. Mary Moncrieff-Yates and Neil E. Rigler, ibid. 3 
385 (1953). 

1294 Thiomycin, golden yellow needles, m.p. 176-178° (dec). 

Resembles thioaurin somewhat. May be identical. 
Analysis: C 49.61, H 5.50, N 8.88, S 16.26. Negative 
FeCl,, ninhydrin, Fehling. 



Pfizer Handbook of Microbial Metabolites 6io 

Streptomyces sp. resembling S. phaeochromogenes var. 
chloromyceticus 

Yorio Hinuma, Susumu Hamada, Takaaki Yashima and 
Kyoko Ishikara, /. Antibiotics (Japan) 8A 118 (1955). 

1295 Totomycin, C2iH;.90iiN, amorphous. 

Streptomyces crystallinus 

Jacques Loewe Research Foundation, Inc., British Patent 

758,276 (1956). 

1296 Toyocamycin, C12H14O4N-,, colorless needles, prisms (mono- 

hydrate), m.p. 243°. 

Analysis: Negative FeClg, Fehling, Mohsch, Millon, 
Sakaguchi, EhrUch. Mol. Wt. 286, 266. 

Streptomyces toyocaensis 

Kg Kikuchi, J. Antibiotics (Japan) 8A 145 (1955). 

Haruo Nishimura, Ken Katagiri, Kozaburo Sato, Mikao 
Mayama and Noburo Shimaoka, ibid. 9A 60 (1956). 

1297 Tubercidin, C11H14O4N4, crystals, m.p. 247° (dec). 

Forms a picrate, reineckate, helianthate, and penta- 
chlorophenolate. A basic substance stable to acid and 
alkali. 

A streptomycete 

Kentaro Anzai, Goto Nakamura and Saburo Suzuki, /. Anti- 
biotics (Japan) lOA 201 (1957). 

1298 Unclassified Compound, Ci^HioO.No, m.p. 220° (dec). 

Contains two enolic groups. U.V. bands at 243 and 
374 m/x. Photosensitive. 

Penicillium puherulum (mycelium) 

A. H. Campbell, M. E. Foss, E. L. Hirst and J. K. N. Jones, 
Nature 155 141 (1945). 

1299 Unnamed antibiotic, CnHiyO^N, hygroscopic light yellow crys- 

tals, m.p. 195° (dec). 

U.V. absorption at 365, 410 m^u,. 

Proteus immunitatis anticarcinomatosa n. sp. (on a 
special blood plasma-bouillon medium) 

Atsuo Ushiyama and Takaaki Miyasaka, Japanese Patent 
Application 3998 (1957). 

1300 Vancomycin (Hydrochloride), amphoteric white solid, Mol. Wt. 

3200-3500 ±200 (titr.). 

Streptomyces orientalis n. sp. 



6ii Unclassified Metabolites 

M. H. McCormick, W. M. Stark, G. E. Pittenger, R. C. Pit- 
tenger and J. M. McGuire, "Antibiotics Annual 1955-1956," 
Medical Encyclopedia, Inc., New York, p. 606. 

H. Nishimura, Ann. Kept. Shionogi Res. Lab. 1 479 (1957). 

1301 Variotin, Ci.sH^jyOiN, colorless oil, [a],r' -5.68° (c 1.0 in meth- 

anol). 

A neutral oil with an ester-like odor. C 67.35, H 8.58, 
N 4.16, contains no halogen, sulfur or phosphorus. Posi- 
tive diazo, nitroalkyl and hydroxamic acid reactions; nega- 
tive ferric chloride, Millon, Ehrlich, Sakaguchi, Molisch, 
biuret, xanthoprotein and ninhydrin tests. 

Paecilomyces variotis Bainier var. antibioticus 
Hiroshi Yonehara, Setsuo Takeuchi, Hakao Umezawa and 
Yusuke Sumiki, /. Antibiotics (Japan) 12A 109, 195 (1959). 

1302 Vengicide, C04H09O9N10, white, amorphous, m.p. 241.5-243°, 

[a]i>-" —51.6° (in 0.1 N hydrochloric acid solution). 

Mol. Wt. -600. U.V. Amax. 233.5 and 273.5 m^x in 0.05 
N hydrochloric acid. C 47.05, H 4.85, O 24.85, N 23.85. 

Streptomyces vendargensis 

Oxytetracycline is produced also in this fermentation. 

N. V. Koninklijke Nederlandsche Gist — en Spiritus — fabriek, 
British Patent 764,198 (1956). (Chem. Abstr. 51 10009a) 

A. P. Struyck, Canadian Patent 514,164 (1955). 

1303 Vertimycin C, crystalline, m.p. 152-155°. C 62.4, H 6.84, O 

21.9, N 8.0. 

Streptomyces verticillatus 
Canadian Patent 575,235 (1959). 

1304 Violacetin, fine yellow needles, m.p. (hydrochloride) >210°. 

Basic compound. Positive ninhydrin, diazo tests. Pre- 
cipitated from aqueous solution by picric acid, phospho- 
tungstic acid, forms reineckate. Analysis: C 38.26, H 
6.74, N 24.71, CI 9.33. Negative biuret, Fehling, ninhy- 
drin, glucosamine, maltol, Sakaguchi, Millon, xanthopro- 
tein. 

Streptomyces sp. resembling S. purpurochromogenes 
Kazuyoshi Aiso, Tadashi Aral, Ichiro Shidara, Hiroo Kurihara 
and Yoshiro Morita, /. Antibiotics (Japan) 8A 33 (1955). 

1305 Violarin, C22-24H.i:j .S4OS-9, dark violet color or amorphous red 

powder, dec. 130°, somewhat similar to litmocidin, rubidin 
and rhodomycetin. 



Pfizer Handbook of Microbial Metabolites 612 

Streptomyces violaceus 

N. A. Krasilnikov, G. K. Skryabin and O. I. Artamonova, 
Antibiotiki (U.S.S.R.) 3 (1958). 

Idem., J. Antibiotics (Japan) 13A 1 (1960). 

D. M. Trakhtenberg, L. V. Cerenkova and A. S. Chochlov, 
Symposium on Antibiotics, Prague (1959). 

Viridins, CigHjgOe (isomers). 

1306 a-Viridin, fine colorless needles, m.p. 208-217° (dec), [a]D^° 

-213.4° (in chloroform). 

1307 yS-Viridin, Fine colorless needles, m.p. 140° (dec), [ajn^" -50.7° 

(in chloroform). 

Both compounds show: negative Schiff, FeCly, iodo- 
form. Red- violet color with phloroglucinol-hydrochloric 
acid. Positive ketone derivative tests, Fehling, Tollens. 

Tricho derma viride 

P. W. Brian and J. C. McGowan, Nature 156 144 (1945). 

P. W. Brian, P. J. Curtis, H. G. Hemming and J. C. McGowan, 
Ann. Appl. Biol. 33 190 (1946). 

E. B. Vischer, S. R. Rowland and H. Raudnitz, Nature 165 
528 (1950). 

1308 Virtosin, C27H40O9N2, colorless needles, m.p. 142.5-143°, [aW^ 

+80° ± 0.5° (c 1 in acetone). 

Positive Fehling and Sakaguchi reactions; negative 
ninhydrin and maltol tests. 

Streptomyces olivochromogenes 

Akira Miyake, Shozo Wada, Motoo Shibata, Koichi Naka- 
sawa, Jujo Kaneko and Yasuharu Mamiya (to Takeda Pharma- 
ceutical Industries Ltd.), Japanese Patent Appl. 6149 (1957). 

1309 Wortmannin, colorless needles, m.p. 240° (yellowing in sun- 

light). 

A neutral antifungal antibiotic, containing only C, H, O. 
Yields were about 100 mg. per liter. 

Penicillium wortmanni Klocker 

P. W. Brian, P. J. Curtis, H. G. Hemming and G. L. F. Nor- 
ris, Brit. Mycol. Soc. Trans. 40 365 (1957). 

1310 Xanthicin, C].3H]-,0-,N, yellowish silky crystals, m.p. 211-213° 

(dec), [cxW +319° (c 0.25 in acetone). 

U.V. maxima at 270 m^ (CH3OH). 260 m^, 325 nifx 
(0.1 MKOH). Positive aldehyde, indole, FeClg tests. 
Negative amino, nitro, Fehling's, phosphomolybdic acid 
tests. Alkaline KMn04 oxidation gives succinic acid. 



6i3 Unclassified Metabolites 

Streptomyces xanthochromo genes 

Yasuji Sekizawa and Keiko Miwa, Nippon Noget-kagaku 
Kaisld 30 471 (1956). 

1311 Xanthomycin-like Antibiotic, C29n420-N,)S4Cr (Reineckate), yel- 

low-orange glass, U.V. 264.5, 335 m^^ in water, pH 2. 

Positive Benedict, bromine, silver nitrate, potassium 
iodide, sodium hydrosulfite and periodic acid. 

Streptomyces sp. 

James D. Mold and Quentin R. Bartz, /. Avi. Chem. Soc. 72 
1847 (1950). 

1312 Xanthomycins (Protomycins), C23H09.31O7N3, free base: deep 

orange-red amorphous solid. Dihydrochloride : bright 
orange-yellow plates, [a]ir^ -fll5° (c 0.4 in water). 
Reineckate: long, orange needles, m.p. 165-170° (dec). 

Contains components A and B. Acid hydrolysis yields 
ethanolamine, methylamine and ammonia. Red-purple 
color with alkali. Positive Bro uptake, Benedict, silver 
nitrate, sodium hydrosulfite, ketone derivatives. Negative 
ninhydrin, Molisch, Sakaguchi, FeCls. 

Streptomyces sp. 

C. B. Thorne and W. H. Peterson, /. Biol. Chem. 176 413 
(1948). 

K. V. Rao and W. H. Peterson, /. Am. Chem. Soc. 76 1335 
(1954). 

1313 Xanthothricin, yellow needles, m.p. 165° (s. 161-162°). 

Analysis: C 43.64, H 3.82, N 35.21, O 17.34. 

Streptomyces sp. similar to S. albus 

Roy A. Machlowitz, W. P. Fisher, Betsey S. McKay, Al- 
fred A. Tytell and Jesse Charney, Antibiotics and Chemo- 
therapy 4 259 (1954). 



BIBLIOGRAPHY, REVIEWS AND 
GENERAL REFERENCES 

A book closely related to this one in intent and format is 
Walter Karrer's "Konstitution und Vorkommen dcr organischen 
Pflanzenstoffe (exclusive Alkaloide)." This lists over 2600 
compounds with simple physical properties and thorough refer- 
encing. The emphasis is on metabolites of higher plants, al- 
though many fungal products are listed. 

Another related book is "Type Reactions in Fermentation 
Chemistry," by Lowell L. Wallen, Frank H. Stodola and Rich- 
ard W. Jackson. Here the emphasis is on non-sugar substrates, 
and classification is by type of reaction (oxidation, reduction, 
etc.) accomplished. Many microbial transformations of ster- 
oids are included, for example. Structural formulas, names of 
microorganisms and references are listed. 

The revised edition of W. W. Umbreit's "Metabolic Maps" 
should be mentioned. This is essentially a list of equations, 
outlining various metabolic pathways, with no discussion and 
little referencing, but including catabolic routes and those in 
higher organisms. 

"Naturally Occurring Quinones," by R. Thomson, is similar 
in method to our handbook, but is confined to the single class 
of compounds with more thorough discussion of each entry. 
"The Comparative Biochemistry of the Carotenoids" by T. W. 
Goodwin is somewhat similar in its restriction to a single class 
of chemicals. Both books are broader in scope as far as pro- 
ducing organism is concerned, and are not limited to micro- 
organism products. 

"The Chemistry of Microorganisms," by Arthur Bracken, is 
descriptive in style, showing some of the degradations and 
syntheses leading to establishment of chemical structures and 
offering essays on related topics. There is, perhaps, some 
emphasis on substances isolated and characterized by the 
Raistrick group. 

We have not designated antibiotics as such nor have we 
attempted to separate the commercial from the non-commercial 
or to give the trade names or the biological properties. Data 
on biological properties are difficult to evaluate and, on the 
newer antibiotics, may conflict. Trade names tend to change 
due, for example, to improvements in dosage forms. 



Pfizer Handbook of Microbial Metabolites 6i6 

Many antibiotic spectra as well as physical properties and 
references are given in the "Handbook of Toxicology, Vol. II, 
Antibiotics" edited by W. S. Spector. 

The "Physicians' Desk Reference" is an annual publication 
listing antibiotics and other medicines by brand name, by man- 
ufacturer and by type of medicine. There is also a therapeutic 
indications index, Usting medicines available for the treatment 
of a given condition, and an index Usting professional informa- 
tion (composition, dosage, etc.) on each product. 

The "Antibiotics Annual" series also is a useful reference 
work on antibiotics. 

Various other monographs, reviews and general references 
are in the hst below. 

1 "Konstitution und Vorkommen der orpanischen PflanzenstofFe (ex- 
clusive Alkaloide)," Walter Karrer, Birkhauser Verlag, Basel, 1958, 
1207 pp. An index similar in intent to this book, but with its scope 
the entire plant kingdom. Thoroughly referenced. 

2 "Type Reactions in Fermentation Chemistry," L. Wallen, F. Stodola 
and R. Jackson, Agricultural Research Service, United States De- 
partment of Agriculture (ARS-71-13), Peoria, 1959. A compilation 
of the types of chemical conversions by microorganisms which have 
been reported in the literature with emphasis on non-sugar sub- 
strates. 

3 "Metabolic Maps," W. W. Umbreit, Burgess Publishing Co., Minneap- 
olis, 1960. 

4 "The Chemistry of Microorganisms," Arthur Bracken, Pitman and 
Sons, London, 1955, 343 pp. 

5 "Antibiotics and Mold Metabolites," a symposium at the March 26, 
1956 meeting of the English Chemical Society. Reprinted as Special 
Publication No. 5. 

6 "Chemical Compounds Formed from Sugars by Molds," B. Gould, 
Scientific Report Series No. 7 of the Sugar Research Foundation, 
New York, 1947. 

7 "The Microbes Contribution to Biology," Albert J. Kluyver and C. van 
Niel, Harvard University Press, Cambridge, 1956, 182 pp. 

8 "Industrial Fermentations," Leland A. Underkofler and Richard J. 
Hickey, Chemical Publishing Co., Inc., New York, 1954, Vol. I, 565 
pp.. Vol. II, 578 pp. 

9 Industrial and Engineering Chemistry Annual Unit Process Review 
of Fermentation, Samuel C. Beesch and G. M. Shull, Ind. and Eng. 
Chem. 48 1585 (1956). These reviews list, among other things, 
new antibiotics and new microbiological transformations of steroids. 

10 Industrial and Engineering Chemistry Annual Unit Processes Review 
of Fermentation, Samuel C. Beesch and G. M. Shull, Ind. and Eng. 
Chem. 49 1491 (1957). 



6 1 7 General References 

n Industrial and Enqiveering Chemistry Avnnal Unit Process Review 
of Ferjuentation, Samuel C. Beesch and Fred W. Tanner, Jr., Ind. 
and Eng. Chein. 50 1341-1354 (1958). 

12 Biochemistrii of microorganisms, C. B. van Niel, Ann. Rev. Biochem. 
12 551-586 (1943). A review with 371 references. 

13 "Handbook of Toxicology, Vol. II, Antibiotics," W. S. Spector (Ed.), 
W. B. Saunders and Co., Philadelphia, 1957. This is a compilation 
of data on physical and biological properties of 340 antibiotics or 
substances which have been tested as antibiotics. Most of these are 
microorganism metabolites. Thoroughly referenced. This compila- 
tion was prepared under the direction of the Committee on the 
Handbook of Biological Data, Division of Biology and Agriculture, 
the National Academy of Sciences, The National Research Council. 

14 Chemistry and biochemistry of antibiotics, E. B. Chain, Ann. Rev. 
Biochem. 27 167-212 (1958). A review with 297 references. 

15 Structural chemistry of actinomycetes antibiotics, Eueene van Tame- 
len, Fortschr. Chem. org. Naturstoffe 16 90-138 (1958). A review 
with 113 references. 

16 Biochemistry of antibiotics, S. B. Binkley, Ann. Rev. Biochem. 24 
597-626 (1955). A literature survey complete to October 1954 
with 284 references. 

17 Biochemistry of antibiotics, B. Duggar and V. Singleton, Ann. Rev. 
Biochem. 22 459-496 (1953). A review of the literature to Novem- 
ber 1952 with 288 references. 

18 "Biochemistry of Some Polypeptide and Steroid Antibiotics," CIBA 
Lectures in Microbial Biochemistry, E. Abraham, John Wiley and 
Sons, Inc., New York, 1957. 

19 "Topics in Microbial Chemistry," Antimycin, Coenzyme A, Kinetin 
and Kinins, E. R. Squibb Lectures on Chemistry of Microbial Prod- 
ucts, F. M. Strong, John Wiley and Sons, Inc., New York, 1957. 

20 Antibiotics produced by fungi, P. Brian, Botan. Rev. 17 357—431 
(1951). A review with 276 references. 

21 Antibiotics produced by actinomycetes, R. Benedict, Botan. Rev. 19 
229-320 (1953). A review with 251 references. 

22 "The Phvsicians' Desk Reference (to Pharmaceutical Specialties and 
Biologicals)," 14th Ed., Medical Economics, Inc., Oradell, N. J., 1960. 

23 "Lectures in Antibiotics," G. F. Cause, Medgiz, Moscow, 1959, 356 
pp. (In Russian) 

24 "Antibiotics," Milos Herold, Czechoslovakian Academy of Science, 
Prague, 1957, 363 pp. (In Czechoslovakian) 

25 "New Antibiotic Binan (Usnic Acid)," Symposium on usnic acid 
and its use as an antibiotic. (In Russian) Academy of Science. 
U.S.S.R., 1957, 224 pp. 



Pfizer Handbook of Microbial Metabolites 6i8 

26 "Streptomycin and Dihydrostreptomycin," Louis Weinstein and 
N. Joel Ehrenkranz, Antibiotic Monographs No. 10, Medical Ency- 
clopedia Inc., New York, 1958, 111 pp. 

27 "Streptomycin, Nature and Practical Applications," Selman A. Waks- 
man (Ed.), The Williams and Wilkins Co., Baltimore, 1949, 612 pp. 

28 "Polymyxin, Neomycin, Bacitracin," Ernest Jawetz, Antibiotic Mono- 
graphs No. 5, Medical Encyclopedia Inc., New York, 1956, 85 pp. 

29 "Antibiotics Derived from Bacillus Polymyxa," (a symposium) Roy 
Waldo Miner (Ed.), Annals of the New York Academy of Sciences, 
51, 853-1000 (1949). 

30 "Terramycin, Review of the Literature," Chas. Pfizer and Co., Inc., 
1953, 76 pp. 

31 "Terramycin, Oxytetracycline," Merle M. Musselman, Medical Ency- 
clopedia Inc., New York, 1956, 141 pp. 

32 "Terramycin" (a symposium) Roy Waldo Miner (Ed.), Annals of 
the New York Academy of Sciences, 53, 223-459 (1950). 

33 "Tetracycline," Harry F. Dowling, Antibiotics Monographs No. 3, 
Medical Encyclopedia Inc., New York, 1955, 57 pp. 

34 "A Review of the Clinical Uses of Aureomycin," Lederle Laboratories 
Div., American Cyanamid Co., 1951, 241 pp. 

35 "Aureomycin, Chlortetracycline," Mark H. Lepper, Medical Encyclo- 
pedia Inc., New York, 1956, 149 pp. 

36 "Chloromycetin, Chloramphenicol," Theodore E. Woodward and 
Charles L. Wisseman, Jr., Antibiotics Monographs No. 8, Medical En- 
cyclopedia Inc., New York, 1958, 152 pp. 

37 "Erythromycin," Wallace E. Herrell, Antibiotics Monographs No. 1, 
Medical Encyclopedia Inc., New York, 1955, 56 pp. 

38 "Penicillin," Harold L. Hirsh and Lawrence E. Putnam, Antibiotics 
Monographs No. 9, Medical Encyclopedia Inc., New York, 1958, 
144 pp. 

39 "Antibiotics, A Survey of Penicillin, Streptomycin, and Other Anti- 
microbial Substances from Fungi, Actinomyces, Bacteria, and 
Plants," H. W. Florey, E. Chain, N. G. Heatley, M. A. Jennings, A. G. 
Sanders, E. P. Abraham and M. E Florey, Oxford University Press, 
London, 1949, Vol. I, 628 pp.. Vol. II, 1662 pp. 

40 "Neomycin, Its Nature and Practical Application," Selman A. Waks- 
man (Ed.), The Wilhams and Wilkins Co., Baltimore, 1958, 396 pp. 

41 "The Fifth Year of Aureomycin," Lederle Laboratories Div., Ameri- 
can Cyanamid Co., 1952, 374 pp. 

42 "Antibiotics," Robertson Pratt and Jean Dufrenoy, J. P. Lippincott 
Co., Philadelphia, 2nd. ed., 1953, 369 pp. 



Gig General References 

43 "Antibiotics and Antibiotic Therapy," Allen E. Hussar and Howard L. 
HoUey, Macmillan Co., New York, 1954, 463 pp. 

44 "Chemistry of Proteins," Shiro Akabori (Ed.), Chap. 9, Antibiotic 
Polypeptides, Kyoritsu Shuppan, Tokyo, 1957. (In Japanese) 

45 "Physiology of Fungi," Vincent W. Cochrane, John Wiley and Sons, 
Inc., New York, 1958, 524 pp. Particularly pertinent is Chapter 2 
(pp. 35-55), Tlie Composition of Fungus Cells. 

46 "Chemical Activities of Fungi," Jackson W. Foster, Academic Press, 
New York, 1949, 648 pp. 

47 Chemistry of the Fungi, C. Stickings and H. Raistrick, Ann. Rev. 
Biochem. 25 225-256 (1956). A review with 182 references. 

48 Chemistry of the Fungi, J. Birkinshaw, Ann. Rev. Biochem. 22 371- 
399 (1953). A review with 152 references. 

49 Biochemistry of Fungi, Edward L. Tatum, Ann. Rev. Biochem. 13 
667-704 (1944). A review with 333 references. 

50 Biochemistry of the Lower Fungi, Harold Raistrick, Ann. Rev. 
Biochem. 9 571-592 (1940). A review with 95 references. 

51 Oxygen Heterocyclic Fungal Metabolites, W. Whalley, Prog, in Org. 
Chem. 4 72-113 (1958). 

52 "Essays in Biochemistry," Samuel Graff (Ed.), Some metabolic prod- 
ucts of basidiomycetes, M. Anchel, John Wiley and Sons, Inc., New 
York, 1957, pp. 1-13. A review with 40 references. 

53 "Organic Acid Production by some Wood-Rotting Basidiomycetes," 
G. Walter, Univ. Microfilms Publ. No. 10,417, 99 pp. Dissertation 
Abstracts 15 321 (1955). 

54 "Chemistry of Lichen Substances," Y. Asahina and S. Shibata, Japan 
Society for the Promotion of Science, Tokyo, 1954, 240 pp. 

55 Chemistry of Lichens, Carl Axel Wachtmeister, Svensk Kent. Tidskr. 
70 117-133 (1958). A review in English with 74 references. 

56 Chemical Constitution and Antibiotic Action of Lichen Substances, 
Josef Klosa, Pharmazie 8 435^42 (1953). A review with 59 refer- 
ences. 

57 Algal Chemistry, B. Wickberg, Svensk Kem. Tidskr. 71 87-106 
(1959). A review in English with 73 references. 

58 "The Chemistry and Chemotherapy of Tuberculosis," E. Long, The 
Williams and Wilkins Co., Baltimore, 3rd ed., 1958. 

59 The Chemistry of the Lipids of the Tubercle Bacillus and Certain 
Other Microorganisms, R. J. Anderson, Fortschr. Chem. org. Natur- 
stoffe 3 145-302 (1939). 

60 Chemistry of Bacterial Lipids, J. Asselineau and E. Lederer, Fortschr. 
Chem. org. Naturstoffe 10 170-256 (1953). A review with 362 ref- 
erences; E. Lederer, Angew. Chem. 72 372 (1960). (A review) 



Pfizer Handbook of Microbial Metabolites 620 

61 "Bacterial Fermentations," H. Barker, John Wiley and Sons, Inc., 
New York, 1956, 90 pp. 

62 "Bacterial Anatomy," Sixth Symposium of the Society for General 
Microbiology, E. Spooner and B. Stocker (Eds.), Cambridge Univer- 
sity Press, Cambridge, 1956, 360 pp. 

63 The (phenazine) bacterial pigments, in "Phenazines," George A. 
Swan and Desmond G. I. Felton, Interscience Publishers, New York, 
1957, pp. 174-209. 

64 Structure and Synthesis of Naturally Occurring Polypeptides, F. Rob- 
inson, /. Pharm. and Pharmacol. 8 297-308 (1956). A review with 
89 references. 

65 "Biochemistry of the Amino Acids," Alton Meister, Academic Press, 
New York, 1957. 

66 Paper Chromatographic Investigation of the Amino Acid Content of 
a Variety of Bacterial Hydrolysates, I. Kandler and C. Zehender, 
Arch, filr Mikrobiol. 24 41-48 (1956). (Semiquantitative) 

67 Bacterial and fungal products containing amino sugars, P. W. Kent 
and M. W. Whitehouse, in "Biochemistry of the Amino Sugars," But- 
terworths, London, 1955, pp. 133-161. 

68 Branched Chain Sugars of Natural Occurrence, F. Shafizadeh, Ad- 
vances in Carbohydrate Chemistry 11 263-283 (1956). 

69 Bacterial Dextrans, M. Stacey and C. Ricketts, Fortschr. Chem. org. 
Naturstoffe 8 28-43 (1951). 

70 Die natUrlich vorkommenden Polyacetylen-Verbindungen, F. Bohl- 
mann, Angew. Chem. 67 389 (1955). 

71 Natural Alkynes, J. Beer, Wiadomosci Chem. 9 460^81 (1955). A 
review with 74 references. 

72 Acetylenverbindungen im Pfianzenreich, F. Bohlmann and H. Mann- 
hardt, Fortschr. Chem. org. Naturstoffe 14 45-53 (1957). 

73 Occurrence of Acetylenic Compounds in Nature, P. Wailes, Revs. 
Pure and Appl. Chem. (Australia) 6 61-98 (1956). A review with 
tabulation of ultraviolet absorption data and 89 references. 

74 Acetylenic Compounds as Natural Products, J. Bu'Lock, Quart. Rev. 
10 371-394. A review with 102 references. 

75 Carotenoids, T. W. Goodwin, Ann. Rev. Biochem. 24 497-522 (1955). 

76 Carotenoids in fungi, bacteria and algae, in "The Comparative Bio- 
chemistry of the Carotenoids," T. W. Goodwin, The Chemical Pub- 
lishing Co., New York, 1954, pp. 99-155. 

77 Some Biochemical Aspects of Fungal Carotenoids, F. Haxo, Fortschr. 
Chem. org. Naturstoffe 12 169-197 (1955). A review with 116 ref- 
erences. 



621 General RefercHces 

78 The Biosynthesis and Function of the Carotenoid Pigments, T. W. 
Goodwin, Advances in Enzymology 21 295-361 (1959). 

79 "Naturally Occurring Quinones," R. Thomson, Butterworths, Lon- 
don, 1958. Literature covered through 1956. 

80 Occurrence and Biochemical Behavior of Quinones, O. Hofmann- 
Ostenhof, Fortschr. Chem. org. Naturstoffe 6 159-224 (1950). 

81 Anthraquinone Pigments Produced by Molds, Shoji Shibata, Kagaku 
(Science) 26 391-396 (1956). A review with 41 references. 

82 Tetracyclic Triterpenes, E. Jones and C. Halsall, Fortschr. Chem. 
org. Naturstoffe 12 68-96 (1955). 

83 Chlorine Containing Metabolic Products, I. Yoshida, Kagaku no 
Ryoiki (J. Japan. Chem.) 5 406-409, 419 (1951). A review with 19 
references. 

84 Vitamins in Microorganisms, J. Van Lanen and F. W. Tanner, Jr., 
Vitamins and Hormones 6 163-224 (1948). A review with 361 ref- 
erences. 

85 "Special Publication No. 12 of the English Chemical Society," 1958, 
especially The Biosynthesis of Aromatic Compounds from C, and C, 
Uiiits, A. J. Birch and Herchel Smith, pp. 1-13, and Biosynthesis of 
Aromatic Ring Systems from C^ and C, fragments, Gosta Ehrensvard, 
pp. 17-31. 

86 "The Structural Relations of Natural Products," R. Robinson, Oxford 
University Press, London, 1955, 150 pp. 

87 Biosynthetic Relations of Phenolic and Enolic Compounds, A. J. 
Birch, Fortschr. Chem. org. Naturstoffe 14 186-216 (1957). 

88 "Perspectives in Organic Chemistry," Alexander Todd (Ed.), Inter- 
science Publishers, New York, 1956, especially Biosynthetic Theories 
in Organic Chemistry, A. J. Birch, pp. 134—155, and Microorganisms 
in Organic Chemistry, Karl Folkers, pp. 392-430. 

89 A Region of Biosynthesis, H. Raistrick, Proc. Roy. Soc. A 199 141- 
168 (1949). A review of fungal metabolites with 157 references. 

90 Microbiological Conversions of Steroids, Drurey H. Peterson, Record 
Chem. Prog. 17 211-240 (1956). 

91 Transformations of Steroids by Molds, Gilbert Shull, Trans. N. Y. 
Acad. Sci. 19 147-72 (1956). A review with 63 references. 

92 Microbiological Alterations of Steroids, P. Enthoven, Chem. Weekblad 
52 166-172 (1956). A review with 40 references. 

93 Microbiological Conversions of Steroids for Technical Purposes, 
E. Vischer and A. Wettstein, Angew. Chem. 69 456-463 (1957). A 
review with 70 references. 

94 Enzymic Transformations of Steroids by Microorganisms, E. Vischer 
and A. Wettstein, Adv. Enzymol. 20 237-282 (1958). A review. 



Pfizer Handbook of Microbial Metabolites 622 

95 "Chemical Transformations by Microorganisms," F. Stodola, John 
Wiley and Sons, Inc., New York, 1958, 134 pp. 

96 The oxidation of aromatic rings by microorganisms in metabolism, 
F. Happold in Biochemical Symposium No. 5, "Biological Oxidation 
of Aromatic Rings," R. T. Williams (Ed.), 1950. A review with 46 
references. 

97 The use of biochemical oxidations and reductions for preparative 
purposes, F. Fisher, "Newer Methods of Preparative Organic Chem- 
istry," Interscience Publishers, Inc., New York, 1948, pp. 159-196. 

98 Allgemeine Methoden zur Ausfiihrung biochemischer Reaktionen, 
B. Helferich, H. Stetter and J. Krebs, "Methoden der organishen 
Chemie," Georg Thieme Verlag, Stuttgart, 1955, Band IV, pp. 822- 
902. 



APPENDIX A 



The Chemical Composition of the Tissues 
and Large Molecules of Bacteria and Fungi 



The composition of the cell wall, the capsule and the proto- 
plast membrane in bacteria and of the mycelial wall in molds 
is generally more specific to the organism than that of the 
lower molecular weight metabolites. For that reason these 
substances are more interesting in taxonomy and immuno- 
chemistry. The toxins, pyrogens and lipoproteins are also in- 
teresting from these standpoints. 

The advent of paper chromatography has so facilitated the 
identification of amino acids, sugars and other fragments of 
the hydrolysis of the higher molecular weight components of 
microorganisms that the literature on this topic has blossomed 
during recent years. 

Some of the results have been unexpected. For example, 
the actinomycetes, which resemble the molds superficially, 
have been found closer chemically to the bacteria. 

This appendix is a list of references on the subject. While 
the paper titles may not always so indicate, they are all con- 
cerned in some way with the composition or structure of the 
tissues and macromolecules of bacteria and fungi. 

Pastenrella septica (P. multocida). I. The occurrence of type- 
specific polysaccharides containing aldoheptose sugars. 
A. P. MacLennan and C. J. M. Rondle, Nature 180 1045 (1957). 

Specific polysaccharide of Pasteurella pestis. 
D. A. L. Davies, Biochem. J. 63 105 (1956). 

Natural occurrence of a new aldoheptose sugar. 
D. A. L. Davies, Nature 180 1129 (1957). 

Elemental and amino acid composition of purified plague toxin. 
D. F. Bent, H. Rosen, S. M. Levenson, R. B. Lindberg and Samuel J. 
Ajl, Proc. Soc. Exptl. Biol. Med. 95 178 (1957). 

Role of a,e-diaminopimelic acid in the cellular integrity of Escher- 
ichia coli. 
Lionel E. Rhuland, J. Bacterial. 73 778 (1957). 

A colicin from Escherichia coli SG710. 
Rainer Niiske, Gottfried Hosel, Harry Venner and Helmut Zinner, 
Biochem. Z. 329 346 (1957). 



Pfizer Handbook of Microbial Metabolites 624 

An agent from Escherichia coli causing hemorrhage and regression 
of an experimental mouse tumor. IV. Some nitrogenous components 
of the phospholipid moiety. 

Miyoshi Ikawa, J. B. Koepfli, S. G. Mudd and Carl Niemann, }. Am. 
Chem. Soc. 75 3439 (1953). 

Colominic acid, a polymer of N-acetylneuraminic acid. 
Guy T. Barry, J. Exp. Med. 107 507 (1958). 

Capsular polysaccharides of Escherichia coli types K28A and 
K34A. 
BiU B. Wiley and Henry W. Scherp, Can. J. Microbiol. 4 505 (1958). 

The chemical and serological relationships of certain polysac- 
charides containing sialic acid. 

Guy T. Barry, Tien-Hu Tsai and Francis P. Chen, Nature 185 597 
(1960). 

Structure of the capsular polysaccharide of Aerobacter aerogenes 
(NCTC 418). 

S. A. Barker, A. B. Foster, I. R. Siddiqui and M. Stacey, /. Chem. Soc, 
2358 (1958). 

The extracellular polysaccharide of Aerobacter aerogenes A3 (Sj). 
J. F. Wilkinson, W. F. Dudman and G. O. Aspinall, Biochem. J. 59 
446 (1955). 

Chromatographic analysis of hydrolysates of Salmonella typhosa. 

F. Savoia, Boll. soc. ital. biol. sper. 32 226 (1956). 

Chemical composition of Salmonella antigen II. Chemical com- 
position of antigen O of Salmonella kirkei and Salmonella hvitting- 
foss. 

G. Bo, A. Defranceschi and G. C. Nava, Giom. microbiol. I 247 
(1955). 

Contributions to the study of the antityphi-paratyphi vaccines II. 
A comparative chemical study of the somatic antigens of Salmonella 
typhi (S. typhosa) extracts. 

E. Soru, C. Barber, S. Toma, V. Gritaenco and B. Bogokowski, Acad, 
rep. populare Romine, Studii cercetari chim. 4 243 (1956). 

The biological action of highly purified pyrogens (lipopolysac- 
charides) from Salmonella ahortivoequina. 

E. Eichenberger, M. Schmidhauser-Kopp, H. Hurni, M. Fricsay and 
O. Westphal, Schweiz. med. Wochschr. 85 1190, 1213 (1955). 

The hexose constituents of some shigella polysaccharide hydroly- 
zates. 

D. A. R. Simmons, J. Gen. Microbiol. 17 650 (1957). 
Epidemiology of Shigella sonnei. I. Biochemical characteristics. 

Szymona Szturm-Rubensten and Danielle Piechaud, Ann. inst. Pas- 
teur 92 335 (1957). 

The chemical constitution of brucella. 

E. M. Gubarev, E. K. Alimova and G. D. Bolgova, Biokhimiya 21 647 
(1956). 

The specific polysaccharides of some gram-negative bacteria. 
D. A. Davies, Biochem. J. 59 696 (1955). 



625 Appendix A. 

The chemistry and biochemistry of typhoid antigens. 
A. De Barbicri. Atti Congr. intern, standard Immunomicrobiol. 
(Rome) 2 257 (1956). 

Toxic end-products from Pasteurella pestis. II. Toxin yields as 
influenced by conditions of growth. 
K. Goodner, /. Infectious Diseases 97 246 (1955). 

Studies on plague. I. Purification and properties of the toxin of 
Pasteurella pestis. 

Samuel J. Ajl. Jeanette S. Reedal, E. L. Durram and Joel Warren, 
;. Bacterial. '70 158 (1955). 

Isolation of a polysaccharide from Vibrio fetus. 
S. M. Dennis, Nature 183 186 (1959). 

Chemical investigation of the endotoxin of Pseudomonas aerugi- 
nosa. 

Fugio Egami, Michio Shimomura, Hiroshi Ishihara, J. Y. Homma, 
K. Sagehashi and Seigo Hosoya, Bull. soc. chim. biol. 36 779 (1954). 

Rhamnose and rhamnolipide biosynthesis by Pseudomonas aerugi- 
nosa. 

George Hauser and Manfred L. Karnovsky, J. Biol. Chem. 224 91 
(1957). 

Chemical studies on endotoxins I. Chemical composition of the 
endotoxin of Shigella flexneri 2B. 

Chiaki Nishimura, Masao Nakamura, Reiko Ofuchi, Shigeo Iwahara 
and Yasuhiko Nozaki, Japan. J. Microbiol. 2 179 (1958). 

Toxins of Pseudomonas pseiidomallei. II. Characterization. 
Robert J. Heckly and Clara Nigg, /. Bacterial. 76 427 (1958). 

Occurrence of poly-/3-hydroxybutyric acid in aerobic gram-negative 
bacteria. 

W. G. C. Forsyth, A. C. Hayward and J. B. Roberts, Nature 182 800 
(1958). 

Sulla composizione chimica degli antigeni delle salmonelle. Nota 
III. Composizione chim. degli antigeni O delle Salmonelle tel aviv 
cholerae suis e montevideo. 
G. C. Nava, G. Bo and A. Defranceschi, Giorn. m.icrobiol. 4 95 

(1957). 

Characterization of intracellular glucosidic polysaccharide pro- 
duced by Brucella suis. 

N. D. Gary, L. L. Kupferberg and L. H. Graf, /. Bacteriol. 76 359 
(1958). 

A group of pseudomonads able to synthesize poly-/3-hydroxybutyric 
acid. 
M. B. Morris and J. B. Roberts, Nature 183 1538 (1959). 

Production of a mannose polysaccharide by Pseudomonas fluores- 
cens from low molecular weight sources. 
Robert Garfield Eagen, Dissertation Abstr. 20 477 (1959). 

Composition of cell walls of variants of Salmonella typhimurium. 
M. Herzberg, J. H. Green and J. C. Boring, Bacteriol. Proc, 169 
(1960). 



Pfizer Handbook of Microbial Metabolites 626 

Enterotoxin. 
Kikuo Fujiwara and Tetsujiro Sugiyama, Nippon Saikingaku Zasshi 
10 189 (1955). 

Polyribophosphate, the type-specific substance of Hemophilus 
influenzae , type B. 

Stephen Zamenkof, Grace Leidy, Patricia L. Fitzgerald, Hattie E. 
Alexander and Erwin Chargaff, /. Biol. Chem. 203 695 (1953). 

The polysaccharide produced by Azotobacter indicum. 
Clara M. Quinnell, S. G. Knight and P. W. Wilson, Can. J. Microbiol. 
3 277 (1957). 

Extracellular polysaccharides of rhizobium. 
Beverly A. Humphrey and J. M. Vincent, /. Gen Microbiol. 21 477 
(1959). 

The isolation of D-fucosamine from the specific polysaccharide of 
Chromobacterium violaceum (NCTC 7917). 
M. J. Crumpton and D. A. L. Davies, Biochem. J. 70 729 (1958). 

A galactan from Mycoplasma mycoides. 
P. Plackett and S. H. Buttery, Nature 182 1236 (1958). 

Circular paper chromatography of long-chain fatty acids in the 
analysis of (gram-negative) bacterial lipopolysaccharides. 
A. Nowotny, A. Liideritz and O. Westphal, Biochem. Z. 330 47 
(1958). 

The extracellular polysaccharide of Xanthomonas phaseoli. 
S. Lesley and R. Hochster, Can. J. Biochem. and Physiol. 37 513 
(1959). 

A function for the extracellular polysaccharide of Azotobacter 
vinelandii. 
Michael H. Proctor, Nature 184 1934 (1959). 

Lipides of the cell envelope of Azotobacter vinelandii. 
Allen G. Marr and Tsuneo Kaneshiro, Bacteriol. Proc, 63 (1960). 

Chemical composition of cell walls of drug resistant neisseriae. 
R. P. Pradhan and W. A. Konetzka, ibid., 170 (1960). 

Amino acids of red sulfur bacteria. 
H. Mukherjee, Nature 184 1742 (1959). 

A new amino sugar present in the specific polysaccharides of some 
strains of Chromobacterium violaceum. 
M. J. Crumpton and D. A. L. Davies, Biochem. J. 64 22p (1956). 

Immunopolysaccharides. XI. Structure of an Acetobacter capsula- 
tum dextran. 

S. A. Barker, E. J. Bourne, G. T. Bruce and M. Stacey, }. Chem. Soc, 
4414 (1958). 

Oligosaccharide formation during synthesis of cellulose by Aceto- 
bacter acetigenum. 

T. K. Walker and H. B. Wright, Arch. Biochem. and Biophys. 69 362 
(1957). 

Bacterial levans of intermediate molecular weight. 
James R. Mattoon, Chester E. Holmlund, Saul A. Schepartz, James J. 
Vavra and Marvin J. Johnson, Appl. Microbiol. 3 321 (1955). 



627 Appendix A. 

The nature of the polysaccharides of the dextran-producing or- 
ganisms Leuconostoc mesenteroides, Leuconostoc dextranicum and 
Streptococcus bovis. 
R. W. Bailey and A. E. Oxford, /. Gen. Microbiol. 20 258 (1959). 

Characterization of dextrans from four types of Leuconostoc 
mesenteroides. 

Allene Jeanes, W. C. Haynes and C. A. Wilham, J. Bacteriol. 71 167 
(1956). 

Characterization and classification of dextrans from ninety-six 
strains of bacteria. 

Allene Jeanes, W. C. Haynes, C. A. Wilham, J. C. Rankin, E. H. Mel- 
vin. Marjorie J. Austin, J. E. Cluskey, B. E. Fisher, H. M. Tsuchiya 
and C. E. Rist, /. Am. Chem. Soc. 76 5041 (1954). 

Cell wall composition of leptotrichia species. 
G. H. G. Davis and A. C. Baird-Parker, Nature 183 1206 (1959). 

Composition of the cell wall of Staphylococcus aureus. Its rela- 
tion to the mechanism of action of penicillin. 

Jack L. Strominger, James T. Park and Richard E. Thompson, J. Biol. 
Chem. 234 3263 (1959). 

The amino acid composition of the protein and cell wall of 
Staphylococcus aureus. 
R. Hancock, Biochim. et Biophys. Acta 37 42 (1960). 

Composition of the cell wall of Staphylococcus aureus 209P. 
Nobutoshi Ishimoto, Masahiro Saito and Eiji Ito, Nature 182 959 
(1958). 

Staphylococcal toxins. III. Partial purification and some proper- 
ties of 5-lysin. 
A. W. Jackson and R. M. Little, Can. J. Microbiol. 4 453 (1958). 

The intracellular amino acids of Staphylococcus aureus: release 
and analysis. 
R. Hancock, Biochim. et Biophys. Acta 28 402 (1958). 

Constitution of a muco-complex of Micrococcus lysodeikticus I. 
Isolation and purification. 

Shichiro Akiya and Otomatsu Hoshino, Yakugaku Zasshi 77 777 
(1957). 

Development of lysozyme resistance in Micrococcus lysodeikticus 
and its association with increased 0-acetyl content of the cell wall. 
W. Brumfitt, A. C. Wardlaw and J. T. Park, Nature 181 1783 (1958). 

Partial acid hydrolysis of the cell wall of Micrococcus lysodeikticus. 
H. R. Perkins and H. J. Rogers, Biochem. J. 69 15p (1958). 

The chemical composition of the protoplast membrane of Micro- 
coccus lysodeikticus. 

A. R. Gilby, A. V. Few and Kenneth McQuillen, Biochim. et. Biophys. 
Acta 29 21 (1958). 

Products of partial acid hydrolysis of mucopeptide from cell walls 
of Micrococcus lysodeikticus. 
H. R. Perkins and H. J. Rogers, Biochem. J. 72 647 (1959). 

The structure of a disaccharide liberated by lysozyme from the cell 
walls of Micrococcus lysodeikticus. 



Pfizer Handbook of Microbial Metabolites 628 

H. R. Perkins, ibid. 74 182 (1960). 

Synthesis of carbohydrates by Micrococcus ureae from acetic acid. 
V. I. Lyubimov, Doklady Akad. Nauk S.S.S.R. Ill 881 (1953). 

The biosynthesis of a streptococcal capsular polysaccharide. 
Yale J. Topper and Murray M. Lipton, /. Biol. Chem. 203 135 (1953). 

The production of capsules, hyaluronic acid, and hyaluronidase by 
twenty-five strains of group C streptococci. 

A. P. MacLennan, J. Gen. Microbiol. 15 485 (1956). 

Variation in the group-specific carbohydrate of group A strepto- 
cocci II. The chemical basis for serological specificity of the carbo- 
hydrate. 
Maclyn McCarty, /. Exp. Med. 104 629 (1956). 

Production of hyaluronic acid in the resting cells of group A 
Streptococcus hemolyticus. 

Seiki Hayano and Haruo Iwasawa, Nippon Saikingaku Zasshi 10 269 
(1955). 

Examination of the L-forms of group A streptococci for the group- 
specific polysaccharide and M protein. 

John T. Sharp, W. Hijmans and L. Dienes, J. Exp. Med. 105 153 
(1957). 

Studies of streptococcal cell walls. IV. The conversion of D-glu- 
cose to cell wall L-rhamnose. 

W. H. Southard, J. A. Hayashi and S. S. Barkulis, J. Bacteriol. 78 79 
(1959). 

Studies of streptococcal cell walls. V. Amino acid composition 
of cell walls of virulent and avirulent group A hemolytic strep- 
tococci. 

B. S. Tepper, J. A. Hayashi and S. S. Barkulis, ibid. 79 33 (1960). 
Studies of streptococcal cell walls. III. The amino acids of the 

trypsin-treated cell wall. 

James A. Hayashi and S. S. Barkulis, ibid. 77 177 (1959). 

Precipitation of the specific polysaccharide of Cryptococcus neo- 
formans A by types II and XIV antipneumococcal sera. 
P. A. Rebers, S. A. Barker, M. Heidelberger, Z. Dische and E. E. 
Evans, /. Am. Chem. Soc. 80 1135 (1958). 

The genus cryptococcus. 
Rhoda W. Benham, Bacteriol. Revs. 20 189 (1956). 

Immunopolysaccharides. VIII. Enzymic synthesis of 6-0-a-D-glu- 
copyranosyl-3-O-methyl-D-glucose by Betacoccus arabinosaceous. 
S. A. Barker, E. J. Bourne, P. M. Grant and M. Stacey, ]. Chem. Soc, 
601 (1958). 

The cell wall of Myxococcus xanthus. 
D. J. Mason and Dorothy Powelson, Biochim. et Biophys. Acta 29 1 
(1958). 

The cell wall of myxobacteria. 
Donald Joseph Mason, Dissertation Ahstr. 18 1949 (1958). 

A rapid and specific method for the isolation of pneumococci 
polysaccharide. 



629 Appendix A. 

A. S. Markowitz and Jack R. Henderson, Nature 181 771 (1958). 

a,e-Diaminopimelic acid metabolism and sporulation in Bacillus 
sphaericiis. 
Joan F. Powell and R. E. Strange, Biochem. J. 65 700 (1957). 

a,e-Diaminopimelic acid in the peptide moiety of the cell wall poly- 
saccharide of Bacillus anthracis. 
H. Smith, R. E. Strange and H. T. Zwartouw, Nature 178 865 (1956). 

The structure of the immunospecific polysaccharide of Bacillus 
anthracis. 
L. Mester and G. Ivanovics, Chem. and Ind., 493 (1957). 

Polyglutamic acid from the capsule of Bacillus anthracis grown in 
vivo; structure and aggressin activity. 

H. T. Zwartouw and H. Smith, Congr. intern, biochim.. Resumes 
Connnuns., 3e Congr. (Brussels), 93 (1955). 

Chemical structure of capsular glutamyl polypeptide of Bacillus 
megaterium and Bacillus anthracis. 

T. Amano, M. Torii, M. Tokuba, O. Kurimura, T. Morishima and 
S. Utsumi, Med. J. Osaka Univ. 8 601 (1958). 

The polysaccharide from Bacillus anthracis grown in vivo. 
H. Smith and H. T. Zwartouw, Biochem. J. 63 447 (1956). 

Polysaccharide containing amino sugar from Bacillus suhtilis. 
Nathan Sharon, Nature 179 919 (1957). 

Isolation of d- and L-glutamyl polypeptides from culture filtrates 
of Bacillus subtilis. 

Curtis B. Thorne and C. Gomez Leonard, /. Biol. Chem. 233 1109 
(1958). 

Physicochemical studies of poly-D-glutamic acid from Bacillus 
anthracis grown in vitro. 

L. H. Kent, B. R. Record and R. G. Wallis, Phil. Trans. Roy. Soc. 
London 250 1 (1957). 

The composition of the spore wall and the wall of the vegetative 
cell of Bacillus subtilis. 
M. R. J. Salton and Betty Marshall, J. Gen. Microbiol. 21 415 (1959). 

The diaminohexose component of a polysaccharide isolated from 
Bacillus subtilis. 
Nathan Sharon and Roger W. Jeanloz, /. Biol. Chem. 235 1 (1960). 

Structure of teichoic acid from the walls of Bacilhis subtilis. 
J. J. Armstrong, J. Baddiley and J. G. Buchanan, Nature 184 248 
(1959). 

Composition of teichoic acids from a number of bacterial walls. 
J. J. Armstrong, J. Baddiley, J. G. Buchanan, A. L. Davison, M. V. 
Kelemen and F. C. Neuhaus, Nature 184 247 (1959). 

Polysaccharide capsule of Bacillus megaterium. 
J. Tomcsik and Joyce B. Baumann-Grace, Proc. Soc. Exp. Biol. Med. 
101 570 (1959). 

Biochemical study of the products (polysaccharides) from a grow- 
ing aerobic bacterium: Bacillus megatherium. 
J. P. Aubert, Ann. Inst. Pasteur 80 644 (1951). 



Pfizer Handbook of Microbial Metabolites 630 

The chemical nature of the cytoplasmic membrane and ceU wall 
of Bacillus megaterium, strain M. 

C. WeibuU and L. Bergstrom, Biochim. et Biophys. Acta 30 340 
(1958). 

The polysaccharide produced by Bacillus polymyxa X. Component 
sugars. 

Akira Misaki, Toshihiko Higashi and Shiro Teramoto, Hakko Kogaku 
Zasshi 36 181 (1958). 

Studies on the bacterial polysaccharide, rhamnogalactan, produced 
by Bacillus polymyxa var. XI. 

Akira Misaki, Yoshiaki Yagi and Shiro Teramoto, /. Fermentation 
Technol. 36 25 (1958). 

A mannan produced by Bacillus polymyxa. 

D. H. BaU and G. A. Adams, Can. J. Chem. 37 1012 (1959). 
Polysaccharide produced by a strain of Bacillus polymyxa. 

Akira Misaki and Shiro Teramoto, J. Fermentation Technol. 36 266 
(1958). 

Isolation and chemical nature of capsular and cell wall haptens 
in a bacillus species. 
S. Guex-Holzer and J. Tomcsik, J. Gen. Microbiol. 14 14 (1956). 

Bacterial ceU wall. XIII. Chemical composition of bacterial cell 
wall and spore membranes. 

Nagayuki Yoshida, Yoshifumi Izumi, Isamu Tani, Saburo Tanaka, 
Kenji Takaishi, Tadayo Hashimoto, Komei Fukui and Chiaki Furu- 
kawa, Tokushima J. Exptl. Med. 3 8 (1956). (Chem. Ahstr. 51 
13054f) 

Isolation and structure of ribitol phosphate derivatives (teichoic 
acids) from bacterial cell walls. 

J. J. Armstrong, J. Baddiley, J. G. Buchanan, B. Carss and G. R. 
Greenberg, J. Chem. Soc, 4344 (1958). 

Paper chromatographic investigation of the amino acid composi- 
tion of different bacteria hydrolysates. 
O. Kandler and C. Zehender, Arch. Mikrobiol. 24 41 (1956). 

Nucleotides and the bacterial cell wall. 
J. J. Armstrong, J. Baddiley, J. G. Buchanan and B. Carss, Nature 181 
1692 (1958). 

The action of fluorodinitrobenzene on bacterial cell walls. 
V. M. Ingram and M. R. J. Salton, Biochim. et Biophys. Acta 24 9 
(1957). 

Chemical analysis elucidating the structure of bacterial L-forms. 
Claes Weibull, Acta Pathol. Microbiol. Scand. 42 324 (1958). 

The interpretation and use of bacterial infrared spectra. 
Eric K. Rideal and D. M. Adams, Chem. and Ind., 762 (1957). 

Bacterial pyrogens. 
Ivan L. Bennett, Jr. and Leighton E. Chiff, Pharmacol. Rev. 9 427 
(1957). 

The preparation of bacterial pyrogenic substance and its clinical 
application. III. The effects of bacterial pyrogenic substance and of 



63 1 Appendix A. 

the antitumor substances upon Yoshida sarcoma and ascites carci- 
noma 130. 

Kosaku Aoyama, Fumio Miyazawa, Hiromitsu Kurisu, Sadayosi 
Hatta, Hideo Arai, Yoiti Fujita, Mikio Urabe, Yugaku Sakai and 
Takayoshi Aoki, Eisei Shikenjo Hokoku No. 74 361 (1956). 

Determination by paper chromatography of compounds constitut- 
ing bacterial pyrogens. 

K. Macek and Jaroslava Hacaperkova, Ceskoslov. farm. 7 300 (1958). 
(Chem. Abstr. 52 186461) 

The chemical composition of the bacterial cell wall. 
C. S. Cummins, Intern. Rev. Cytology 5 25 (1956). 

Bacterial capsules and their relation to the cell wall. 
J. Tomcsik, edited by E. T. C. Spooner and B. A. D. Stocker, "Bac- 
terial Anatomy," Cambridge University Press, Cambridge, 1956, pp. 
41-67. 

Bacterial cell walls. 
M. R. J. Salton, ibid., pp. 81-110. 

Bacterial protoplasts; their formation and characteristics. 
C. WeibuU, ibid., pp. 111-126. 

Studies of the bacterial cell wall. II. Methods of preparation and 
some properties of cell walls. 

M. R. J. Salton and R. W. Home, Biochim. et Biophys. Acta 7 177 
(1951). 

The molecular basis of antibody formation. 
Leo Szilard, Proc. Nat. Acad. Sci. U. S. 46 293 (1960). 

Nucleotides and bacterial cell wall components. 
J. Baddiley, Proc. Chem. Soc, 177 (1959). 

Chemistry of bacterial cell walls. 
Friedrich Zilliken, Federation Proc. 18 966 (1959). 

Bacterial fructosans and fructosanases. 
Jakob R. Loewenberg and Elwyn T. Reese, Can. J. Microbiol. 3 643 

(1957). 

Cell wall amino acids and amino sugars. 
M. R. J. Salton, Nature 180 338 (1957). 

Composition of the cell wall of Lactobacillus bifidus. 
C. S. Cummins, Olivia M. Glendenning and H. Harris, Nature 180 
337 (1957). 

The nature of D-alanine in lactic acid bacteria. 
Esmond E. Snell, Norman S. Radin and Miyoshi Ikawa, /. Biol. Chem. 
217 803 (1955). 

Structure of the carbohydrates of the diphtheria bacteria. 
O. K. Orlova and E. P. Efimtseva, Biokhimiya (U.S.S.R.) 21 505 
(1956). 

Carbohydrate composition of the envelope of Corynebacterium 
equi. 

Tadeusz Mierzejewski, Ann. Univ. Mariae Curie-Sklodowska, Lublin- 
Polonia 10 93 (1955). 



Pfizer Handbook of Microbial Metabolites 632 

Fixed lipids of diphtheria microbes. 
E. Alymova, Biokhimiya (U.S.S.R.) 22 933 (1958). 

Chemical composition of antigens of corynebacterium. 
J. Kwapinski, Acta Microbiol. Polon. 6 133 (1957). 

"Bacterial toxins," W. van Heyningen, Blackwell Scientific Publica- 
tions, Oxford, 1950, 128 pp. 

The mutation of Corynebacterium pyogenes to Corynebacterium 
hemolyticum. 

W. L. Barksdale, K. Li, C. S. Cummins and H. Harris, J. Gen. 
Microbiol. 16 749 (1957). 

Fructosides formed from sucrose by a corynebacterium. 
Gad Avigad and David S. Feingold, Arch. Biochem. and Biophys. 
70 178 (1957). 

Structure of the mannan of diphtheria bacteria. 
O. K. Orlova, Biokhimiya (U.S.S.R.) 23 467 (1958). 

Amino acid composition of pure diphtheria toxin and toxoid. 
Tokiya Ito, Hisao Uetake and Teuchi Sasaki, Igaku to Seibutsugaku 
26 49 (1953). 

The chemical constituents of Mycobacterium paratuberculosis. 
A. Larsen and R. Merkal, Am. Rev. Tuberc. Pulmonary Diseases 77 
712 (1958). 

Nature of the specific polysaccharides of tubercle bacilli. 
J. Foldes, Acta Microbiol. Acad. Sci. Hung. 4 43 (1957). 

Polysaccharide components of the tubercle bacillus. 
M. Stacey, CIBA Foundation Symposium on Exptl. Tuberc. Bacillus 
and Host, 55 (1955). 

Lipides of human avirulent strain H37Ra of Mycobacterium tuber- 
culosis. 
Jean Asselineau, Bull. soc. chim. biol. 38 1397 (1956). 

Constituents of a toxic lipide obtained from Mycobacterium tuber- 
culosis. 

H. Bloch, J. Defaye, E. Lederer and H. Noll, Biochim. et Biophys. 
Acta 23 312 (1957). 

Proteins of various mycobacteria. I. Chemical properties of sev- 
eral peptides isolated from tuberculin protein obtained from the 
human strain Aoyama-i3. 

Isaku Kasuya, Jinsaku Goto, Sadako Hirai, Taichi Someya and Akira 
Hagitani, Japan. J. Med. Sci. and Biol. 9 93 (1956). 

Proteins of various mycobacteria. II. Chemical properties of sev- 
eral peptides isolated from the tuberculin protein of the bacterial 
cells of human strain Aoyama-j3 and Frankfurt. 
Sadako Hirai, ibid. 9 179 (1956). 

Immunologic significance of the cell wall of mvcobacteria. 
Edgar Ribi, Carl L. Larson, Robert List and William Wicht, Proc. 
Soc. Exp. Biol. Med. 98 263 (1958). 

The constitution of a lipoid-bound polysaccharide from Mycobac- 
terium tuberculosis (Human strain). 

Norman Haworth, P. W. Kent and M. Stacey, J. Chem. Soc, 1220 
(1948). 



633 Appendix A. 

Constitution of mycobacteria. II. Amino acid composition of 
bacterial cells belonging to various species of mycobacteria. 
A. Hirsch, C. Cattaneo and M. Morellini, Giorn. biochim. 6 296 
(1957). 

The composition of the waxes of Mycobacterium marianum. 
Georges Mickel, Compt. rend. 244 2429 (1957). 

Chemistry of some native constituents of the purified wax of 
Mycobacterium tuberculosis. 
Hans Noll, J. Biol. Chem. 224 149 (1957). 

Amino acid composition of mycobacteria. 
Ben Ginsberg, Sarah L. Lovett and Max S. Dunn, Arch. Biochem,. and 
Biophys. 60 164 (1956). 

Amino acid composition of extracellular protein from six myco- 
bacteria. 

Sarah L. Lovett and Max S. Dunn, Proc. Soc. Exp. Biol. Med. 97 
240 (1958). 

Electrophoretic and chromatographic studies on extracts of tu- 
bercle bacilli. 
G. Dragoni and E. Bozzetti, Boll. soc. ital. biol. sper. 32 894 (1956). 

The characterization of mycobacterial strains by the composition 
of their lipide extract. 

D. W. Smith, H. M. Randall, M. M. Gastambide-Odier and A. L. 
Koevoet, Ann. N. Y. Acad. Sci. 69 145 (1957). 

Lipide of bacillus Calmette-Guerin (BCG). I. Glyceryl monomy- 
colate in wax C fraction of the lipide of BCG. 
Torn Tsumita, Japan. J. Med. Sci. and Biol. 9 205 (1956). 

Formation of e-aminosuccinyllysine from e-aspartyllysine from 
bacitracin A and from the cell walls of lactobacilli. 
D. L. Swallow and E. P. Abraham, Biochem. J. 70 364 (1958). 

A comparison of cell wall composition in nocardia, actinomyces, 
mycobacterium and propionibacterium. 
C. S. Cummins and H. Harris, J. Gen. Microbiol. 15 ix (1956). 

The occurrence of O-methyl ethers of rhamnose and fucose in spe- 
cific glycolipides of certain mycobacteria. 

A. MacLennan, D. Smith and H. Randall, Proc. Soc. Biochem. 74 
3 p. (England) (1959). 

Isolation of two difFerent cell wall polysaccharides from tubercle 
bacteria. 
J. Foldes, Natunvissenschaften 46 432 (1959). 

The biochemistry of the actinomycetales. Studies on the cell wall 
of Streptomyces fradiae. 

Antonio H. Romano and Walter J. Nickerson, /. Bacteriol. 72 478 
(1956). 

Comparative biochemical studies of the cell walls of actinomy- 
cetales. 
Arthur Sohler, Dissertation Abstr. 17 2410 (1957). 

Polysaccharide fractions of Actinomyces globisporus streptomy- 
cini. 



Pfizer Handbook of Microbial Metabolites 634 

A. N. Belozerskii and I. B. Naumova, Doklady Akad. Nauk S.S.S.R. 
115 957 (1957). 

The chemical composition of the actinomycetales: Isolation of a 
polysaccharide containing D-arabinose and D-galactose from Nocardia 
asteroides. 
C. T. Bishop and F. Blank, Can. J. Microbiol. 4 35 (1958). 

A lipopeptide from Nocardia asteroides. 
Micheline Guinand, Georges Michel and Edgar Lederer, Compt. rend. 
246 848 (1958). 

Polysaccharide fractions of Actinomyces rimosus and Actinomyces 
aureofaciens. 

A. N. Belozerskii and I. B. Naumova, Doklady Akad. Nauk S.S.S.R. 
122 441 (1958). 

Studies on the cell wall composition and taxonomy of actinomy- 
cetales and related groups. 
C. S. Cummins and H. Harris, /. Gen. Microbiol. 18 173 (1958). 

Biochemistry of the actinomycetales. III. Cell wall composition 
and the action of lysozyme upon cells and cell walls of the actino- 
mycetales. 

Arthur Sohler, Antonio H. Romano and Walter J. Nickerson, J. 
Bacteriol. 75 283 (1958). 

Fluorescence of the toxin of Clostridium botulinum and its rela- 
tion to toxicity. 
Daniel A. Boroff and John E. Fitzgerald, Nature 181 751 (1958). 

Botulinus toxin A and hemaglutinin-receptors of erythrocytes. 

E. R. Gold and I. Ilian, Zentr. Bakteriol. Parasitenk., Abt. I, Orig. 
167 386 (1957). (Chem. Abstr. 51 10655c) 

Polysaccharides isolated from Clostridium perfringens Type C. 

F. Meisel-Mikolajczyk, Bull. acad. polon. sci. 7 213 (1959). (Chem. 
Abstr. 54 1650g) 

Toxin production of Clostridium botulinum (type E). III. Char- 
acterization of toxin precursor. 
Genji Sakaguchi and Sumiko Sakaguchi, J. Bacteriol. 78 1 (1959). 

Isolation of a lipopolysaccharide from Vibrio fetus. 
S. M. Davis, Nature 183 186 (1959). 

Amino acids -and N-terminal groups of scarlatinic toxin (Erythro- 
genic toxin). 

Eugenia Soru, M. Sternberg and M. Istrati, Acad. rep. populare 
Romine, Studii cercetari chim. 5 213 (1957). 

Purification of fiagella and fiagellin by ammonium sulfate. 
Henry Koffler and Toshio Kobayashi, Arch. Biochem. and Biophys. 
67 246 (1957). 

Structure of bacterial flagella. 
W. T. Astbury, E. Beighton and C. WeibuU, Symposia Soc. Exptl. 
Biol. No. 9, Fibrous Proteins and Their Biological Significance, 282 
(1954). 

The cell wall of yeasts — electron miscroscope and X-ray diffraction 
study. 



635 AppendixA. 

A. L. Houwink and D. R. Kreger, Antonie van Leeuwenhoek J. 
Microbiol. Serol. 19 1 (1953). 

Observations on cell walls of yeasts and some other fungi by X-ray 
diffraction and solubility tests. 

D. R. Kreger, Biochim. et Biophijs. Acta 13 1 (1954). 
Cell wall mannan protein of bakers' yeast. 

G. Falcone and Walter J. Nickerson, Science 124 272 (1956). 

On the composition of zymosan. 
Frederick J. DiCarlo and Joseph V. Fiore, Science 127 756 (1958). 

Chemical structure and serologic properties of the polysaccharide 
of Candida albicans. 

Ludwik Rzucidlo, Danuta Weyman, Aleksandra Stachow and Geno- 
wefa Rzesa, Med. Doswiadczalna i Mikrobiol. 7 315 (1955). 

Mucous substance around the cell wall of yeasts. VIII. Polysac- 
charide isolated from filtrate of yeast. 
Tomojiro Kaibara, }. Agr. Chem. Soc. Japan 28 259 (1954). 

Yeast mannan, a cell wall constituent of bakers' yeast. 
P. A. Rollofsen, Biochim. et Biophijs. Acta 10 477 (1953). 

Composition and structure of yeast cell walls. 
A. L. Houwink, D. R. Kreger and P. A. Rollofsen, Nature 168 694 
(1951). 

The composition of fungus cells. 
Vincent W. Cochrane, "Physiology of Fungi," John Wiley and Sons, 
Inc., New York, 1958, pp. 35-55. 

Glucomannan-protein complexes from cell walls of yeasts. 
Gian Kessler and Vv^alter J. Nickerson, }. Biol. Chem. 234 2281 

(1959). 

Isolation and chemical composition of zymosan. 

E. N. Voluiskaya, N. V. Cheburkina, V. I. Tovarnitskii and I. N. 
Nikolskaia, Voprosij Med. Khim. 5 143 (1959). 

Quantitative estimation of chitin in fungi. 
Harold J. Blumenthal and Saul Roseman, /. Bacterial. 74 222 (1957). 

Chitin 
M. V. Tracey, Rev. Pure Appl. Chem. (Australia) 7 1 (1957). 

Chemical composition of the cell wall of Haplo sporangium parvum. 

F. Blank, J. Histochem. and Cytochem. 5 500 (1957). 

The amino acid composition of fusarium mycelium. 

C. S. Venkata Ram, Proc. Natl. Inst. Sci. India Pt. B., 22 227 (1956). 

A qualitative comparison of the amino acid and sugar content of 
acid hydrolysates from the mycelium of several anthracnose fungi. 

D. F. Crossan and D. L. Lynch, Phytopathology 48 55 (1958). 

The chemical composition of the cell walls of dermatophytes. 
F. Blank, Biochim. et Biophys. Acta 10 110 (1953). 

The protein nature of trichophytins and their amino acid compo- 
sition. 

Jacques Meyer, Rene Sartorv, Jacques Malgras and Jacques Touillier, 
Compt. rend. 234 2224 (1952). 



Pfizer Handbook of Microbial Metabolites 636 

The cell walls of dimorphic fungi causing systemic infections. 
F. Blank, Can. J. Microbiol. 1 1 (1954). 

Extracellular metabolic products (polysaccharides) of a hirsutella 
species. 
T. C. Loughheed and D. M. MacLeod, Nature 182 114 (1958). 

An extracellular polysaccharide from Mucor racemosus. 
L. Hough and M. B. Perry, Biochem. J. 61 viii (1955). 

The chemical composition of the mycelium of Penicillium chryso- 
genum. 
J. Janicki and J. Skupin, Acta Microbiol. Polon. 7 139 (1958). 

The amino acid composition of the mold bodies of Aspergillus 
oryzae and Penicillium chrysogenum. 

J. Datta, K. Bhatacharya and D. Roy, Ann. Biochem. and Exp. Med. 
(India) 17 35 (1957). 

A new oligosaccharide (fungitetraose) formed from sucrose. 
Humio Kurasawa, Yukimasa Yamoto, Ikuo Igaue and Yasuchi Naka- 
mura, /. Agr. Chem. Soc. Japan 30 696 (1956). 

The structure of a trisaccharide synthesized by action of Peni- 
cillium chrysogenum on sucrose. 
Alessandro Ballio and Serena Russi, Gazz. chim. ital. 86 476 (1956). 

A chemical and physical investigation of the cell walls of some 
marine algae. 

J. Cronshaw, A. Myers and R. D. Preston, Biochim. et Biophys. Acta 
27 89 (1958). 

Algal (polysaccharide) chemistry. 
B. Wickberg, Svensk Rem. Tidskr. 71 87 (1959). 

Structure of lichenin. 
Stanley Peat, W. J. Whelan and J. G. Roberts, /. Chem. Soc, 3916 
(1957). 

Pathways for biosynthesis of a bacterial capsular polysaccharide. 
Walter H. Taylor, Jr., Dissertation Abstr. 20 3025 (1960). 

On two new antigens in Staphylococcus aureus. 
Gunnar Haukenes and Per Ceding, Acta Pathol. Microbiol. Scand. 49 
237 (1960). 

Acetylhexosamine compounds released enzymically from Micro- 
coccus lysodeikticus cell walls. I. Isolation and composition of ace- 
tylhexosamine-peptide complexes. 

J. M. Ghuysen and M. R. J. Salton, Biochim. et Biophys. Acta 40 462 
(1960). 

Chemical analysis of the wall of myxobacterial microcysts formed 
in liquid culture. 
Jimmy Clarence Adye, Dissertation Abstr. 21 (1960). 

New polysaccharide gums produced by microbial synthesis. 
Manufacturing Chemist 31 206 (1960). 

Physical and chemical analysis of the endotoxin of Salmonella 
enteritidis. 

Edgar Ribi, Bill H. Hoyer, Kelsey C. Milner, Theodore D. Perrine, 
Carl L. Larson, and Granville Goode, J. Immunol. 84 32 (1960). 

Cell wall composition of lactic acid bacteria. 



637 AppendixA. 

Miyoshi Ikawa and Esmond E. Snell, J. Biol. Chem. 235 1376 (1960). 
The cell wall polysaccharides of Candida albicans: glucan, man- 
nan and chitin. 

C. T. Bishop, F. Blank and P. E. Gardner, Can. J. Chem. 38 869 
(1960). 

Nonulosaminic acid (sialic acid) in protisfs. 
S. Aaronson and T. Lessie, Nature 186 719 (1960). 

Isolation and chemical composition of the cell walls of BCG. 
Shozo Kotani, Toshiyuki Kitaura, Teji Hirano and Akira Tanaka, 
Biken's J. 2 129 (1959). (Chem. Abstr. 54 13252c) 

Studies on the muco-complex of Micrococcus hjsodeikticus. 
S. Akiya and O. Hoshino, Chem. and Pharm. Bull. 8 395 (1960). 

Polysaccharides produced by some wood-rotting fungi. 
Francis H. Millazzo, Dissertation Abstr. 21 (1960). 

Cell wall mucopeptides of Staphylococcus aureus and Micrococcus 
lysodeikticus. 
H. Rogers and H. Perkins, Nature 184 520 (1959). 

Purification and chemical properties of flagellin. 
T. Kobavashi, J. Rinker and H. Koffler, Arch. Biochem. and Biophys. 
84 342 (1959). 

Constitution of a glucomannan from wheat stem rust (Puccinia 
graminis tritici) uredospores. 

N. Prentice, L. S. Cuendet, W. F. Geddes and F. Smith, /. Am. Chem. 
Soc. 81 684 (1959). 

Structure of a reserve polysaccharide (leucosin) from Ochro- 
monas malhamensis. 
A. Archibald, D. Manners and J. Ryley, Chem. and Ind., 1516 (1958). 

Polysaccharides of bakers' yeast. II. Yeast glucan. 
S. Peat, W. J. Whelan and T. E. Edwards, J. Chem. Soc, 3862 
(1958). 

Mannan structure of diphtheria bacteria. 

0. Orlova, Biokhimiya 23 502 (1958). 

Uridine diphosphate N-acetylamino sugar compounds from Staph- 
ylococcus aureus strain 209 p. I. Amino acid constituents. 
E. Ito, N. Ishimoto and M. Saito, Arch. Biochem. and Biophys. 80 
431 (1959). 

Amino acid composition of Brazilian Mycobacterium tuberculosis 
BCG strain. 

1. Krzeczkowski and J. Iskierko, Med. Doswiadczalna i Mikrobiol. 9 
185(1957). (Chem. Abstr. 52 17 392a) 

Studies of streptococcal cell walls. III. The amino acids of the 
trypsin-treated cell wall. 
A. Hayashi and S. Barkulis, /. Bacteriol. 77 777 (1959). 

Pigmentation and cell wall material of a Daldinia concentrica 
specimen. 

D. Allport and J. Bu'lock, /. Chem. Soc, 4090 (1958). 
Bacterial endotoxins. 

Otto Westphal, Abstracts 138th Meeting, American Chemical Society, 
New York, September 1960, p. 33-0. 



APPENDIX B 



Bacterial and Fungal Carotenes 



The subject of bacterial and fungal carotenoids is confusing 
because of the large number of closely related structures and, 
in some cases, duplications in nomenclature. The following 
tables were prepared by an authority. Professor T. W. Goodwin 
of the University of Liverpool. They appeared in his excellent 
book "The Comparative Biochemistry of the Carotenoids" and 
are reproduced here with his permission and with the consent 
of the Chemical Pubhshing Co., 212 Fifth Ave., New York City. 



References 




ujpoqjoinjot 


+ + + 


suajndjndopoqj 


+ 


(ui|dopoi|j JO) 

Ujl{4UOX03X| 


+ 4- 


Uj3SO|OjAOpOI{J 


+ + 


ujqiUDxiqnj 


+ 


3U3|njO| 


++ + ++++ 


ausoiAqd | -|- -{- 


auan«o«Xi|d -|- ^_ 4--|_ -f 


3u9jodsojnau 


e»- -j- -{- M- 


auadoaXi + ++ ++ + 


SUSJOJOD-j 


+ 


sua40JD3-g 1 -^- 


^u^^OJD^-^ 


+++++ +++ ++ + 


auaioJD3-£/ + + 4-+ ++ + + + 4-4- + + + + 


aus40JD3-» 1+ + + + ++ + + 


c 

4) 

E 
.? 


Aleuria auranfia 
Allomyces orbuscu/o 
Allom /ces javanicus 
Allomyces macrogyna 
Allomyces moniliformis 
Canfharellus cibarius 
Canfharellus cinnabarinus* 
Canfharellus infunadibiliformis 
Canfharellus lufescens 
Coleosporium senecionis 
Dacromyces sfillafus 
Gymnosporangium /unipenV/rg/n/onoe 
Lycogola epidendron 
Neurosporo crosso 
Phycomyces b/okes/eeonus 
P//obo/us kleinii 
Polystigma rubrum 
Puccinia coronifera 
Rhodoforula rubra 
Rhodoforula sanniei 
Sporobolomyces roseus 
Sporobolomyces salmonicolor 
Tremella mesenferica 









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641 



Appendix-B. 



TABLE II 
Characteristic Fungal Caroienoids* 







Absorption spectra maxima (m^u) 




Melting 
point 








Pigment 


Carbon 
disulphide 


Light 
petroleum 


Chloroform 


Torulene'' * 


185° 


563-5, 520-5, 
488-91 




539,501,469 


Torulcrhodln- 


201-203° 
(decomp.) 


582,541,502 


537,501,467 


554,515,483 


Neurosporene' 










(See also Tetra- 


124° 


502.5, 470.5, 


470,441.5 




hydrolycopene) 




439.5 






Acid carotenoid' 










from Neurosporo crassa 


— 


512-514 


516, 482 




Pigment III 1 from Corfinarius 


— 


— 


520, 470 


462, 405 


Pigment Vlj c/nnabor/nus' — 


494 


— 


455 


Canthoxonthln 218° 


500 


— 


462 



' Pigments first reported in other organisms but also present in fungi are not recorded here. 

References to Table 11 

^E. Lederer, Bull. soc. chim. biol. 20 611 (1938). 

2 P. Karrer and J. Rutschmann, Helv. Chim. Acta 29 355 (1946). 

3 F. Haxo, Arch. Biochem. 20 400 (1949). 
*Idem., Botan. Gaz. 112 228 (1950). 



Pfizer Handbook of Microbial Metabolites 



642 



TABLE III 

Fungi in Which Early Workers''^ Hove Reported fhe Presence of Carofenoids, 

but Which Have Not Recently Been Investigated 



Ascobolus spp. (not A. furfuraceus^) 


Peziza (tochneo) scutellata 


Calocerca cornea 


Phragmidium violaceum 


Calocerca palmata 


Pilobolus crystallimus 


Calocerca viscosa 


Pilobolus kleinii 


Chytridium spp. 


Pilobolus oedipus 


Co/eospoWom Pulsatilla 


Polystigma ochraceum (fulvum) 


Ditiola radicata 


Puccinia coronata 


Leotia lubrica 


Saccharomyces (spp.) 


Lycogola flavofuscunt 


Sphaerostilbe coccaphila 


Melampsora aecidioides 


Spathularia flavida 


Melampsora salicis capreae 


Stemonitis spp. 


Nectria cinnabarina 


Triphragmium ulmariae 


Peziza aurantia 


Uredo [Coleosporium) euphrasie 


Peziza (Lachnum) b/co/or 


Uromyces alchemille 



References to Table III 

1 W. Zopf, "Die Pilze," Trewendt, Breslau, 1890. 
^ F. G. Kohl, Untersuchungen iibei das Carotin und seine physi- 
ologische Bedeutung in der Pflanze, Borntrager, Leipzig, 1902. 
=*T. W. Goodwin, Biochem. J. 50 550 (1952). 



643 



Appendix- B. 



TABLE IV 

Fungi from Which Carotenoids Have Been Shown fo Be Absent 



Agaricus [Telamoria) armillaius^ 


Nephoromo /usifonico' 


Agaricus /oceofus' 


Oidium v/o/oceum' 


AUernaria so/anr'* 


Paxillus ofrofomenfosos' 


Amanifa muscar/o* 


Penicilliopsis clavariaeformis^ 


Amanita pantherina^ 


Peziza aeruginosa^ 


Arthonia spp.' 


Peziza echinosporo' 


Ascobolus furfuracevs^ 


Peziza songuineo' 


Bachospora c/rymo' 


Phragmidium vio/oceum' 


Bacidia moscorum' 


Pichia spp.^ 


Bioforo fungidula^ 


Polyporus grammocephalus^ 


Bilimbia melaena' 


Polyporus /uzonenis* 


Bo/e/us /un'dus' 


Polyporus rubidus^ 


Boletus scober' 


Polyporus zonalis^ 


6ue//ia spp.' 


Polystictus hirsutus^ 


C/odonia coccifero' 


Polystictus sanguineus^ 


Clavaria fern/co' 


Polystictus versicolor' 


C/ov/ceps spp.^ 


Polystictus xanthopus^ 


Cortinarius bulliardi^ 


Pullularia spp.^ 


Cortinarius vio/oceus' 


Rhi'zocfon/a so/Zoni^* 


Daedalea flavida" 


Rhizopogon rufaescens' 


Fusarium iycopersici'* 


Russula alutacea^ 


Fusarium moniforme~ * 


Russula ourafo' 


Fusarium oxysporium-* 


Russu/o emef/co' 


Ganoderma (Fames) lucidus^ 


Russula Integra^ 


Gamphidius glutinasus^ 


Saccobolus vio/oceus' 


Gamphidius viscidus^ 


Sorcogyme pru/noso' 


Helminthosporium sativum^'* 


Taphrina deformans^ 


Helvetia esculenta 


Thalloidima condidum' 


Hydnum ferrugineum^ 


The/ephorus spp.' 


Hydnum repandum^ 


Th/'e/cvio terr/co/o^* 


Hygrophorus cocc/neus^ 


Toru/opsis lipofera'' 


Hygrophorus conicus' 


Torulopsis /uteo/o^ 


Hygrophorus pun/cens' 


Toru/opsis pu/cherrima^'^ 


Lactarius del/ciosus' 


Tramefes persooni* 


Lecidea spp.' 


Tromefes versofi/is^ 


(enzf'fes subferruginea^ 


Zygosacchoromyces spp.^ 



' Only vitamin A-active carotenoids are absent from these species. Inactive carotenoids may possibly 
be present. 

References to Table IV 

1 W. Zopf, "Die Pilze," Trewendt, Breslau, 1890. 
-D. Gottlieb and G. M. Gilligan, Arch. Biochem. 10 163 (1946). 
^T. W. Goodwin, Biochem. J. 50 550 (1952). 

* E. M. Mrak, H. J. Phaff and G. Mackinney, J. Bacteriol. 57 407 
(1949). 

5 S. R. Bose, Trans. Nat. Inst. Sci. India 2 69 (1941). 

^ M. F. Champeau and P. J. Luteraan, Ann. Parasit. 21 344 (1946). 



Pfizer Handbook of Microbial Metabolites 



644 



TABLE V 
Properfies of Bacierial Carofenoids 







Absorption maxima in 


m;u 




Melting 
















Name 


point 


Light 
petroleum 


Carbon 
disulphide 


Chloroform 


Sarcinene^* 


— 


415,440,469 






Sarcinaxanthin^t 


149-150" 


415,440,469 


464,494 


423,451,480 


Xanthophyll^'t (Lutein) from 










Sorc/no lufea 







466,499 


451,480 


Flavorhodene^'^J 


111-113° 


442,470 


472,503 


453,482 


(Rhodoviolacein) 










Rhodopurpurene''^§ 


162° 


472,502 


479,511,550 


458,487,527 


Rhodopin^'^ 


171° 


440,470,501 


478,508,547 


453,486,521 


Rhodovibrin''^ 


168° 




517,556 




Rhodoviolascin^'^ 


218° 




496,530,573.5 


476,507,544 


( = Spirilloxanthin) 










a-Bacteriopurpurin®' H 


— 


460,495,528 
(in methanol) 


498,532,571 




/3-Bacteriopurporin''# 


— 


452,482,502 
(in methanol) 






Leprotene' 


198-200° 


425,452,484 


477,499,517 


428,460,495 


Xanthophyll from 










Flavobacf. esferoaroma- 










iicum, F. suaveoleus and 










F. faecale^ 







453,482,513 


460,513 


Carotene from F. 










sulphureum^* 


— 




437,466,487 


451,481 


Xanthophyll from Erwinia 










laythri^ 


— 




478,513 


458,485 


Xanthophyll from E. ananas^ 


— 




474,508 


460,493 


Chrysophlein^i" 


— 


452 


487 


— 



* The probable identi-'y of these with neurosporene cannot be ignored, 
t These may be identical. 

t May be identical with e-carotene. 

§ May be identical with lycopene. 

II a-Bocteriopurpurin is probably one of Karrer's rhodocarotenoids. 

# ^-Bacteriopurpurin is probably identical with rhodoviolascin. 

References to Table V 

1 E. ChargafF and J. Dieryck Naturzvissenchaften 20 872 (1932). 
- Y. Takeda and T. Ohta, Hoppe-Seyl. Z. 268 1 (1941). 
^B. Sobin and G. L. Stahly, J. Bacteriol. 44 265 (1942). 
* P. Karrer and U. Solmssen, Helv. Chim. Acta 18 25 1306 (1935). 
^ P. Karrer, U. Solmssen and H. Koenig, Helv. Chim. Acta 21 545 
(1938). 

•5H. F. M. Petter, Amsterdam Akad. Wiss. 34 No. 10 (1931). 
^ E. Lederer, Bull. soc. chim. hiol. 20 611 (1938). 

8 Y. Takeda and T. Ohta, Hoppe-Seyl. Z. 262 168 (1939). 

9 G. Turian, Arch. Sci. Soc. Phys. Hist. Nat. Geneve 3 79 (1950). 
^Udem., Helv. Chim. Acta 33 1303 (1950). 



APPENDIX C . 

The Chemical Constituents of Mycobacteria 



A great many metabolites of mycobacteria have been charac- 
terized, many of them incidental to the study of tuberculosis. 
The following referenced list was prepared by Dr. Esmond R. 
Long and appeared in his recent book, The Chemistry and 
Chemotherapy of Tuberculosis. It is reproduced here by per- 
mission of the author and of the Williams and Wilkins Publish- 
ing Co. of Baltimore. While many of the compounds in this 
list appeared earlier in the Handbook, it may be useful to see 
them in aggregate as well. 



< 


■^ 






UJ 


•~ 


y- 




u 




< 

CO 


c 

4> 


O 


•S* 


u 


0) 


>- 




s 


0) 






u. 

O 


.c 


«/) 


V 


t- 




z 


■»; 


m 




D 




K- 








1- 




</5 




7 




o 




u 


<^ 



3 4) 

U E 



O^ r- lO O. 



oo Ij: o- 



CM CN 



- 00 

J^ <3 — 



CO 



O CM 



.— "O 



CM ■— 



CO . CN — . , , . 

O lOlV.O — CSO 

— _.— OCNCO — >OCN 

O . CO . 

•— — O — O 



O T3 

f^ S ^ 2- 8 2 S 



U -D 


D >^ >« o) a ■£ 



Q o ^3 



O O O ^ 



rs» o t^v o 

r^ — ^ <N 

— " — — "" C) 

fv^ O ■>!■ CN 

. — — CN 



>0 -* — — 



— 00 o lo — 



— — CN 



.— <o 


ts. o 




. r^ 


CN CN 




o — 


CM CS 


>o 


^— 




CN CO 


•— 




— 00 



CO •— 



^ o ^ ^ 

crT lo "O cn" ct" o 

O <N 00 o n o 

— CN . — — CN 



S K — K ^ 

^ ■* K 00 O 

-*' o* nT o o" ■* 

O- -^ lO CO O CN 



lO — — — 



O O 4) — <J 

i« »i «» o •- 

O O :5 2 2 

Q. Q. q: I— 13 



4) D) S 

S 2 ^ 



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1^ a. Q. f- => 



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a) £ 
II 
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o .= 



■o 12 



o o 



u — 

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— 3 U 






5 5 5 o o o 






'- 00 



<N r- 





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o» 




00 






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•^ 


^3-."'-. 


^ >o 


nT - o , - ■<«■ 


. CN 


. ^ CO "o >o 


K CM 


>o -o ^ ~ — 



CO -o VO 
■O — <N 
- . CN 
— O . 
■O CO O 



^ 


'■-'^ j; 






«0 CN 


>o 






— K 


"O 


CO ^- CM 








« 


•~ O - 






CO . 


■>t 


- . -* 






>rt in 


CN 


— 00 -o 






— CO 


^ 


r— ^— »— 




cs 


^ ^ 


CO 


^ 


o 


o 


•o — 


CN 


•o 


•— 


•— 


CO yj >o 



^ 2- U i 



c tfc -i: 
a. Q. Q. 



^ O o ^ 



-a 1? _ -i: .bi 

■- >• 4) <J »- 

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■s O. T JP -5 



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K 
















V. 






















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CN 






^ 
















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o ^ 

hs CN 








K 


N, 


<o 


N. 


N, 


K 




00 , . — 00 








CN 


CN 


>o 


CN 


CN 


CN 




— CS o -o — 

CN -O rs ^ CN 












OQ- 
















X 






— "O 










O^ K K 






■c 






— >o 










o — — 






o 
















— CS CN 










rC ". 




K 






^ . . CS 










— -<r 










CO o o o 






►^ 






— n 




■" 


CD 




CS rs hN — 

CS , , , CS . CS 








CO 00 


00 


lO 


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ut" 


_ CO CO 




O — CS o o — o 








lO "O 


>o 


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lO 


Is. 


— lO lO 




^ "O <0 CN ^ lO — 






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CN 
CN 






CN 
CN 


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CN 


CN 




cs 


CN « 


CN 




^ 






CN 


CN 


r— 


CN 


CN -^ 


CS 




o 


<u 






<N 


cs 


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CN 


CN >0 


CN 




CN 


3 













































to 






-* 


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"* 














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o- 




o 


































o 






















CO 


■* k' 


Is." -* 


rC -<t K 


"t K 












— CN 


CN 


o- 


CN 


O- CN 


c^ CN 




o« o 

O CN 








CN 


■t 


rC 


















-«t 


o- 


•— -^ 


















r— 


^- 


•— o 
















c 



E 


^ 




■>* 








<o 






O K 


rv 


'^"" ". 


O 


K 


■^ 




K 






■* CN 


CN 


— lO 




CN 


cs 












r— 


•- •« 




r— 


r— 




CO 






3 


^ , 






CN 


•^ > 


- N. 




~o 






-««• •* 


-«t 


CN "", 


-* 


O IN, 


-tr CN 


o> 


^ 










-^ •— 


o ><» 


r— 




• — >— 


-<1- 


CS — 








w— ^- 


o •- 


•— n 


^ ^ »^ 


f— 


r— 


>0 ^0 








^ , 




^ r— 


r^ 


■o . 


« -* 




CO , 








■* — 


^- 


•o 


CN 


.— >o 


• — • — 


■^ 


— -«t O CN 








-«t -<* 


■t 


K 


■" 


^ 1^ 


-« '- 


•o 


CN N3 CN ^ 




0) 


c 










































3 








o 
















■o 














o 






"5 


(U 






CN 








lO 






U 


E 






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00 






-a 














.£ -a 














_c 














ID 
01 




in 

Q 

u 

< a 


a 


3 
C 



J3 




a 


2 '^ 
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a 

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1- 


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C 

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c 

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c 
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-ri 


c 
Li 


.c 

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. '- CO 

CM lO •— 
O >- CN 

. >o" cn' 

O — — 
O- — CN 



COO ......... 

.— no>ootN<or>-v<o — <o 
00 .»— cNcoooooajcoo-'— •— 

COlO-— •—•— — — — •—•— CNCN 

.o .......... 

CO .(NO«00>O — -t^ooo — ■* 

coo — — CN-^OOOOOOOOO — 

.-•t — — — — — — — — CNfN 

CO 



>? ^ -= 

i o c 
8 O 



CO CO . CO ^ 
-<t >0 CN ^ "O 



E "^ 



CN CN CN 



:° O 



O V 






•I -0 -2 2 



^ .£ -^ 



Pfizer Handbook of Microbial Metabolites 652 

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Pfizer Handbook of Microbial Metabolites 654 

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Pfizer Handbook of Microbial Metabolites 656 

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127. Y. Khouvine and L. Wysmann, Compt. rend. 239 834 (1954). 

128. Y. Kinoshita, Japan J. Tuberc. 3 42 (1955). 

129. W. F. Kirchheimer and C. K. Whittaker, Am. Rev. Tuberc. 70 
920 (1954). 

130. V. Kocher and E. Sorkin, Helv. Chim. Acta 35 1741 (1952). 

131. M. Kusunose, S. Nagai, E. Kusunose, Y. Yamamura, J. Tani, 
T. Terai, T. Nagasuga and Y. Yamamura, Symposia on Enzyme 
Chem. (Japan) 10 114 (1954). 

132. M. Kusunose, E. Kusunose and Y. Yamamura, ibid., 7 59 
(1952). 

133. P. P. Laidlow and H. W. Duldey, Brit. J. Exptl. Pathol. 6 197 
(1925). 

134. S. G. Laland, W. G. Overend and M. Webb, J. Chem. Soc, 3224 
(1952). 

135. A. Lamensans, P. Grabar and J. Bretey, Compt. rend. 232 1967 
(1951). 

136. M. Landy and D. M. Dicken, Proc. Soc. Exp. Biol. Med. 46 449 
(1941). 

137. M. Landy, N. W. Larkum and E. J. Ostwald, ibid. 52 337 (1943). 

138. Z. Lassot, Acta Biochim. Polon. I 239 (1954). 

139. A. Lesuk and R. J. Anderson, J. Biol. Chem. 136 603 (1940). 

140. P. A. Levene, /. Med. Research 6 135 (1901). 

141. M. Lindsay, T. V. O'Donnell and N. L. Edson, Biochem. J. 46 
248 (1950). 

142. E. R. Long, Am. Rev. Tuberc. 4 842 (1921). 

143. A. Lutz, Experientia 3 244 (1947). 

144. M. A. Macheboeuf and M. Faure, Compt. rend. 209 700 (1939). 

145. G. J. Martin, J. Am. Chem. Soc. 60 768 (1938). 

146. E. Marschmann and E. Kiister, Z. physiol. Chem. 193 215 
(1930). 

147. P. Masucci, K. L. McAlpine and J. T. Glenn, Am. Rev. Tuberc. 
22 669, 678 (1930). 

148. M. Maxim, Z. Biochem. 223 404 (1930). 

149. R. L. Mayer, /. Bacteriol. 48 337 (1944). 

150. R. L. Mayer and M. Rodbart, Arch. Biochem. 11 49 (1946). 

151. A. E. O. Menzel and M. Heidelberger, J. Biol. Chem. 124 89, 301 
(1938). 

152. J. Meyer, J. Malgras and R. Schar, Bull, assoc. diplomes micro- 
biol. faculte pharm. (Nancy), 10 (1953). 

153. G. Michel and E. Lederer, Compt. rend. 240 2454 (1955). 

154. G. Middlebrook, Am. Rev. Tuberc. 69 471 (1954). 

155. I. Millman and G. P. Youmans, /. Bacteriol. 69 320 (1955); 
idem., Proc. Soc. Exp. Biol. Med. 91 271 (1956). 



657 Appendix C. 

156. R. C. MiUs. G. M. Briggs, T. D. Luckey and C. A. Elvehjem, 
Proc. Soc. Exp. Biol. Med. 56 240 (1944). 

157. J. H. Mueller, /. Exp. Med. 43 9 (1926). 

158. H. Noll and H. Block. /. Biol. Chem. 214 251 (1955). 

159. H. Noll, H. Block, J. Asselineau and E. Lcderer, Biochim. et 
Biophtis. Acta 20 299 (1956). 

160. K. Ogura, S. Imazu, M. Kato and Y. Yamamura, Kekkaku 
(Tuberculosis) 29 128 (1954). 

161. T. Ohta, ;. Pliarm. Soc. Japan 71 462 (1951). 

162. M. C. Pangborn and R. J. Anderson, /. Biol. Chem. 90 45 
(1931); 94 465 (1931); 101 105 (1933); idein., J. Am. Chem. 
Soc. 58 10 (1936). 

163. M. C. Pangborn, E. Chargaff and R. J. Anderson, /. Biol. Chem. 
98 43 (1932). 

164. R. L. Peck and R. J. Anderson, ibid., 138 135 (1941); 140 89 
(1941). 

165. F. G. Petrick, J. Bacteriol. 48 347 (1944); 51 539 (1946). 

166. F. J. Philpot and A. I. Wells, Am. Rev. Tuberc. 66 28 (1952). 

167. N. Polgar, ;. Chem. Soc, 1008, 1011 (1954). 

168. H. Pope and D. T. Smith, Am. Rev. Tuberc. 54 559 (1946). 

169. S. Raffel, J. Asselineau and E. Lederer, Ciba Found. Symposium 
Exp. Tuberc. (London), 174 (1955). 

170. R. E. Reeves and R. J. Anderson, J. Am. Chem. Soc. 59 858 
(1937); idem., J. Biol. Chem. 119 535 (1937). 

171. A. G. Renfrew, /. Biol. Chem. 83 569, 579 (1929). 

172. E. G. Roberts and R. J. Anderson, ibid. 90 33 (1931). 

173. F. Rohner and F. Roulet, Biochem. Z. 300 148 (1939). 

174. F. F. Rose and G. A. Snow, Ciba Found. Sym,posium. Exp. 
Tuberc. (London), 41 (1955). 

175. A. J. Rosenberg and A. Andrejew, Compt. rend. 235 1437 
(1952). 

176. F. Roulet, H. Wydler and E. A. Zeller, Helv. Chim. Acta 29 1973 
(1946). 

177. F. Roulet and E. A. ZeUer, ibid. 31 1915 (1948). 

178. H. Saito, ;. Biochem. (Japan) 34 223 (1941). 

179. T. Sasakawa, T. Kimura and H. Katayama, Symposia on En- 
zyme Chem. (Japan) 10 103 (1954). 

180. W. Schaefer, Ann. inst. Pasteur 72 783 (1946); 73 749 (1947). 

181. F. B. Seibert, Am. Rev. Tuberc. 17 403 (1928); 59 86 (1949); 
idem., Faraday Soc. Discussion, No. 13 251 (1953). 

182. F. B. Seibert, C. Crumb and M. V. Seibert, /. Am. Chem. Soc. 
72 2678 (1950). 

183. F. B. Seibert and A. M. Fabrizio, Am. Rev. Tuberc. 66 314 
(1952). 

184. F. B. Seibert and J. T. Glenn, ibid. 44 9 (1941). 

185. F. B. Seibert and B. Munday, ibid. 25 724 (1932); 30 713 
(1934). 

186. F. B. Seibert and J. W. Nelson, J. Am. Chem. Soc. 65 272 
(1943). 

187. F. B. Seibert, K. O. Pedersen and A. Tiselius, J. Exp. Med. 68 
413 (1938). 



Pfizer Handbook of Microbial Metabolites 658 

188. F. B. Seibert, E. Soto-Figueroa and E. H. DuFour, Am. Rev. 
Tuberc. 71 704 (1955). 

189. F. B. Seibert, M. Stacey and R. W. Kent, Biochim. et Biophys. 
Acta 3 632 (1949). 

190. F. B. Seibert and D. W. Watson, /. Biol. Chem. 140 55 (1941). 

191. B. Siegel, G. A. Candela and R. M. Howard, J. Am. Chem,. Soc. 
76 1311 (1954). 

192. J. Singer and E. Chrysner, Am.. Rev. Tuberc. 65 779 (1952). 

193. J. D. Smith and G. R. Wyatt, Biochem. J. 49 144 (1951). 

194. G. A. Snow, Congr. intern, biochim. 2' Congr., Resumes 
communs., Paris, 1952, p. 95; idem., J. Chem. Soc, 2588 
(1954). 

195. E. Sorkin and S. V. Boyden, /. Immunol. 75 22 (1955). 

196. E. Sorkin, H. Erlenmeyer and H. Block, Nature 170 124 (1952). 

197. M. A. Spielman, /. Biol. Chem. 106 87 (1934). 

198. M. A. Spielman and R. J. Anderson, ibid. 112 759 (1936). 

199. M. Stacey, P. W. Kent and E. Nassau, Biochim. et Biophys. 
Acta 7 146 (1951). 

200. W. Steenken, Jr., J. Biol. Chem. 141 91 (1941). 

201. N. Stendal, Compt. rend. 198 1549 (1934). 

202. F. H. Stodola and R. J. Anderson, /. Biol. Chem. 114 467 (1936). 

203. F. H. Stodola, A. Lesuk and R. J. Anderson, ibid. 126 505 
(1938). 

204. H. R. Street and R. E. Reeves, Proc. Soc. Exp. Biol. Med. 44 641 
(1940). 

205. W. B. Sutton, /. Biol. Chem. 210 309 (1954); 216 749 (1955). 

206. P. Szafranski, Acta Biochim. Polon. 1 116 (1954). 

207. L. Szarkowska and P. Szafranski, Acta Biochim. Polon. 1 225, 
249 (1954). 

208. J. W. Szarkowski, Bull. acad. polon. sci. class II 2 97 (1954). 

209. M. Szymonowa and O. Sakllawska-Szymonowa, Biokhimiya 19 
295 (1954). 

210. T. Tada and K. Takeya, Japan Soc. for the Promotion of Sci. 
(Tokyo), 1 (1955). 

211. Y. Takeda and T. Hoshino, Japan. J. Tuberc. 2 201 (1954). 

212. Y. Takeda, N. Kasai and Y. Aoki, Japan. J. Exptl. Med. 22 413 
(1952). 

213. Y. Takeda and T. Ohta, J. Pharm. Soc. Japan 64 67 (1944). 

214. K. Takeya and I. Mifuchi, Enzymologia 16 366 (1954). 

215. S. Tamura', Z. physiol. Chem. 87 85 (1913). 

216. D. M. Tennent and D. W. Watson, J. Immunol. 45 179 (1942). 

217. C. M. Todd, Biochem. J. 45 386 (1949). 

218. G. Turian, Helv. Chim. Acta 33 1303 (1950); 34, 1060 (1951). 

219. N. Uyei and R. J. Anderson, J. Biol. Chem. 94 653 (1932); 97 
617 (1932). 

220. S. F. Velick, ibid. 154 497 (1944). 

221. E. Vilkas and E. Lederer, Compt. rend. 240 1156 (1955); idem.. 
Bull. soc. chim. France 38 111 (1956). 

222. E. Vischer, S. Zamenkof and E. Chargaff, /. Biol. Chem. 177 
429 (1949). 

223. W. A. Volk and Q. N. Myrvik, Am. Rev. Tuberc. 73 589 (1956). 



659 Appendix C. 

224. D. W. Watson, Dissertation, University of Wisconsin, 1941. 

225. C. W. Wieghard and R. J. Anderson, /. Biol. Chem. \2<a 515 
(1938). 

226. E. Work, Biochem. J. 19 17 (1951). 

227. Y. Yamamura, Sijmposia on Enzijme Chem. (Japan) 10 114 
(1954). 

228. Y. Yamamura, M. Kusunose and E. Kusunose, Nature 170 207 
(1952); idem., J. Biochem. (Tokyo) 42 785 (1955). 

229. Y. Yamamura, M. Kusunose, S. Nagai, E. Kusunose, Y. Yama- 
mura, J. Tani, T. Terai and T. Nagasuga, Med. J. Osaka Univ. 6 
489 (1955). 

230. Y. Yamamura, K. Ogura and S. Imazu, Symposium on Enzyme 
Chem. (Japan) 8 96 (1953). 

231. Y. Yamamura and S. Watanabe, Kekkaku (Tuberculosis) 27 414 
(1952). 

232. Y. Yamamura, K. Matsui and H. Maeda, /. Biochem. (Tokyo) 
4;} 409 (1956). 

233. E. A. Zeller, L. S. VanOrden, W. F. Kirchheimer and J. C. Laza- 
nas, /. Bacteriol. 67 153 (1954); idem., J. Biol. Chem. 209 429 
(1954). 



REFERENCES TO REVIEW BOOKS AND 

ARTICLES ON CONSTITUENTS OF 

MYCOBACTERIA 

1. R. J. Anderson, Fortschr. Chem. org. Naturstoffe 3 145 (1939); 
idem., Sigma Xi Quart. 27 39 (1939); idem., Harvey Lectures 
35 271 (1939-1940); idem., Chem. Rev. 29 225 (1941); idem., 
Yale J. Biol, and Med. 15 311 (1943). 

2. J. Asselineau, Advances in Tuberc. Research 5 1 (1952). 

3. E. ChargafF and J. N. Davidson, "The Nucleic Acids — Chemistry 
and Biology," Academic Press, Inc., New York, 1955, Vol. I. 

4. W. F. Drea and A. Andrejew, "The Metabolism of the Tubercle 
Bacillus," Charles C. Thomas, Springfield, 111., 1953. 

5. P. Hauduroy, E. Chain, H. Florey, K. A. Jensen, G. Penso and 
J. Trefouel, "Bacilles tuberculeux et paratuberculeux," Masson 
et Cie., Paris, 1950. 

6. P. W. Kent and M. W. Whitehouse, "Biochemistry of the Amino- 
sugars," Butterworth and Co., Ltd., London, 1955. (Also pub- 
lished by Academic Press, Inc., New York, 1955.) 

7. E. Lederer, Proc. Colloq. Chemotherapy Tuberc, Dublin, 1951; 
idem., Congr. intern, biochim., 2" Congr., Symposium sur le 
metabolism microbien, Paris, 1952; idem., Angew. Chem. 72 
372 (1960). 

8. L. Negre, "Les lipoides dans les bacilles tuberculeux et la 
tuberculose," Masson et Cie., Paris, 1950. 

9. F. Roulet and M. Brenner, Zentr. ges. Tuberk. Forsch. 56 193 
(1943). 

10. F. B. Seibert, Chem. Rev. 34 107 (1944); idem., Schweiz. Z. 



Pfizer Handbook of Microbial Metabolites 660 



Tuberk., Separatum, Fasc. 3 1 (1950); idem., Ann. Rev. Micro- 
biology 4 35 (1950). 

11. B. Skowronska-Serafinowa, Wiadomosci Chem. 7 216 (1953). 

12. M. Stacey, Schweiz. Z. Tuberk., Separatum, Fasc. 9 7 (1955). 

13. M. Stacey and R. W. Kent, Advances in Carbohydrate Chem. 3 
311 (1948). 



ADDENDUM 

In order to cover pertinent literature appearing as late as 
December, 1960 this addendum is attached. Also included is 
a little material from earlier dates which was overlooked. Ar- 
rangement is by chapter title, and new compounds eligible for 
inclusion often are given appropriate entry numbers, but with a 
letter added to the number so that it is evident in the indexes 
that such entries are located in the addendum. Due to time 
restrictions these entries may be abbreviated, but references are 
listed. The addendum is not indexed. 

2. Alcohols, Glycols and Compounds Related to Sugars 

17a Acetyl Methyl Carbine! (Acetoin) 

This substance, mentioned as a co-product of butanediol, 
is produced by many microorganisms. It is given off by 
several streptomycetes, including Streptomyces erythreus, 
an erythromycin producer. It is present in such large 
quantities in some erythromycin fermentations that it 
interferes with production of the antibiotic.^ 

A survey has been made of 44 species and strains of 
acetobacter for ability to convert lactate to acetoin.^' A. 
rancens and A. pasteiirianus were good producers, the 
former yielding one isomer, the latter the other. 

Acetoin metabolism of bacteria in general has been 
studied.^" 

Biosynthesis of acetoins has been reviewed.^" 

47a Galactosyl Lactose 

This trisaccharide was produced by Penicillhim chryso- 
genum Thom on a lactose medium and assigned the 
structure 0-/?-D-galactopyranosyl-(l -> 6)-0-/3-D-galacto- 
pyranosyl-(l —> 4)-D-glucopyranose.^ 

Several papers have appeared on the mode of action of 

^ V. Musilek, V. Sevcik, M. Musilkova, J. Rokos and P. Prochazka, 
Experientia 14 323 (1958). 

1" J. de Ley, J. Gen. Microbiol. 21 352-365 (1959). 

i^Yasuhiro Maeda, Okayama Igakkai Zasshi 71 8017 (1959). 
(Chem. Abstr. 55 694i) 

1' H. Oberman, Postepy Biochemii 6 181-195 (1960). 

2 A Ballio and S. Russi, Tetrahedron 9 125 (1960). 



Pfizer Handbook of Microbial Metabolites 662 

streptomycin. Its effect on Escherichia coli has been 
studied.^ The cell permeability barrier was altered, remi- 
niscent of detergents and of polymyxin. Preformed cells 
were undamaged, but defects were caused in cell mem- 
branes formed in its presence by non-resistant cells. 
When C^Mabeled streptomycin was used, initial uptake 
occurred only outside the cell wall and secondary uptake 
depended on secondary damage to the membrane. The 
growing membrane was the primary site of action of the 
antibiotic. 

The effect of streptomycin on the excretion of nucleo- 
tides by E. coli has been investigated.'* Streptomycin en- 
hanced excretion of 5'-nucleotides and prevented excre- 
tion of 2'- or 3'-nucleotides. It was not clear whether 
streptomycin blocked RNA synthesis de novo or whether 
degradation of RNA to 5'-nucleotides was enhanced. 

The same group has published on chloramphenicol-sen- 
sitive and chloramphenicol-insensitive phases of the lethal 
action of streptomycin.'^ It appeared that the lethal ef- 
fect of streptomycin on E. coli was exerted in two phases 
(1) a preparatory phase, which is markedly less lethal 
and can be blocked by chloramphenicol (a protein syn- 
thesis inhibitor), followed by (2) a more direct lethal 
phase which is insensitive to chloramphenicol. The in- 
duction process might have been due to formation of a 
permease without which streptomycin could not accumu- 
late in the cell in lethal concentration. 

It has been found that, while penicillin inhibits growth 
of Staphylococcus aureus (strain Duncan), it does not 
cause rapid lysis as, e.g., in the case of E. coli. Penicillin 
and streptomycin added (each at minimally bactericidal 
concentrations) to exponentially growing cultures caused 
rapid lysis. Only antibiotically active forms of strepto- 
mycin were effective. Under anaerobic conditions lysis 
was not rapid. (Streptomycin is not ordinarily effective 
under such conditions.*^) 



•^Nitya Anand and Bernard D. Davis, Nature 185 22, 23 (1959). 
* Carmen L. Rosano, Richard A. Peabody and Charles Hurwitz, 
Biochim. et Biophys. Acta 37 380 (1960). 

5 Charles Hurwitz and Carmen L. Rosano, ibid. 41 162 (1960). 
6R. Hancock, Nature 186 658 (1960). 



663 



Addendum 



It has been reported that streptomycin inhibits de- 
hydrogenases by influencing the apoenzyme/ The con- 
clusion was made that further search for enzymatic 
reactions susceptible to streptomycin should be aimed at 
the study of its influence on intracellular synthetic proc- 
esses, mainly the synthesis of nucleic acids and proteins. 

The mode of action of streptomycin in connection with 
its binding by Mijcohacterimn avium has been studied.* 

The stereochemistry of neobiosamine B is as shown." 



HO— CH 




Neobiosamine B 



CHi— NH2 



Dextromycin is neomycin B and contains a small 
amount of neomycin C.^" Framycetin also is identical 
with neomycin B." 

59a Aminocidin (Crestomycin, Antibiotic 1600, Pharmiglucin, F. I. 
5853) C03H45OJ4N, (Sulfate) [aW + 51° in water. Pro- 
duced by Streptomyces crestomyceticus, n. sp.^- This 
antibiotic seems to be similar to or identical with paromo- 
mycin. 

" K. Michalska, Symposium on Antibiotics, Prague, 1959. 

^Tatsuji Kinoshita, Nagoija ]. Med. Sci. 21 323 (1958). 

" Kenneth L. Rinehart, Alexander D. Argoudelis, Townley P. Cul- 
bertson, W. Scott Chilton and Klaus Streigler, J. Am. Chem. Soc. 82 
2970 (1960). 

^° Sueo Tatsuoka, Akira Miyake and Hayao Nawa, /. Antibiotics 
(Japan) llA 193 (1958). 

" Kenneth L. Rinehart, Jr., Alexander D. Argoudelis, William A. 
Goss, Arthur Sohler and Carl P. Schaffner, /. Am. Chem. Soc. 82 
3938 (1960). 

^- F. Arcamone, C. Bertazzoli, M. Ghione and T. Scotti, Giorn. 
Microbiol. 7 251 (1959). 



Pfizer Handbook of Microbial Metabolites 664 

D-Araboascorbic acid is produced by Penicillium de- 
cumbens, P. chrysogenum mutant fulvescens, P. notatum, 
P. meleagrinum and P. cyaneofulvum growing on sucrose, 
glucose or D-gluconate." 

3. Aliphatic Acids and Glycolipides 

The name mycoside has been suggested to designate a 
type-specific glycolipide of mycobacterial origin. To 
clarify nomenclature it was proposed that Ca from photo- 
chromogenic strains be called mycoside A, Gr from bovine 
strains, mycoside B, and J.u from avian strains mycoside 
C. Some properties are listed :^* 
Mycoside A: 

Nearly colorless solid, m.p. 105°, [ajn"" — 37° (in 
chloroform). Anal: C 72.2, H 11.3, -OCH, 8.6, N 0.0, 
P 0.0. U.V. maxima at 222, 274, 278 m^ (in hexane). 
Contains 2-0-methylfucose, 2-O-methylrhamnose and 2,4- 
di-0-methylrhamnose. The lipide part is a mycocerosate 
of an aromatic alcohol. 
Mycoside B: 

Colorless wax, m.p. 25°, [a]i.-° — 22° (in chloroform). 
Anal: C 76.6, H 12.0, -OCH3 4.3, N 0.0, P 0.0. U.V. 
maxima at 222, 274, 281 m^. Contains only one sugar, 
2-O-methylrhamnose. The lipide moiety is a diester of 
2 molecules of a branched-chain acid fraction of mean 
molecular weight corresponding to C22H44O2 with a 
phenolic alcohol. It may also sometimes contain myco- 
cerosic acid. 
Mycoside C: 

A peptide-glycolipide mixture. One component sepa- 
rated on sihca gel had the following properties: 
m.p. 200°, [a]D"'-31° (in chloroform). Anal: 
Calculated for C73H133O24N., : C 59.8, H 9.1, N 4.8, 
-OCH3 6.3 

Found: C 60.1, H 8.7, N 5.1, -OCH3 6.0. 
It contains three deoxyhexoses, one being 6-deoxytalose 
and one 3,4-dimethoxyrhamnose. The peptide moiety is 

"T. Takahashi, M. Mitsumoto and H. Kayamori, Nature 188 411 
(1960). 

1* Donald W. Smith, H. M. Randall, A. P. MacLennan and E. Led- 
erer, ibid. 186 887 (1960). 



665 Addendum 

a pentapeptide containing 1 mole of D-phenylalanine, 2 
moles of flf/o-threonine and 2 moles of n-alanine. The 
lipide moiety was not entirely pure, but may be a hydroxy 
acid of about C._.4H4hO:{. Two 0-acetyl groups are present 
in mycoside C. 

The lipoids of mycobacteria, their chemical structures 
and biological effects have been reviewed.^" 

132a IVIycocerosic Acid, C;{2H,.40o, isolated by Anderson and collabo- 
rators,^'^ has been shown to be 2,4,6,8-tetramethylocta- 
cosanoic acid:^" 

CH3— (CH:),9— CH— CHj— CH— CHj— CH— CH>— CH— COOH 
CH3 CH3 CH3 CH3 

Indications were obtained for the presence in mycobac- 
teria of normal chain acids with 22, 24, 26 and 28 carbon 
atoms; 2-, 4-, 6-trimethyl-substituted acids with 25, 27 
and 29 carbon atoms; and 2-, 4-, 6-, 8-tetramethyl-sub- 
stituted acids with 30, 32 and 34 carbon atoms. 

Succinic, fumaric and acetic acids as well as d,l-5- 
carboxymethylhydantoin, shown below, have been iso- 

COOCH3 
/ 
HN — 



O N O 

H 

lated as extracellular acids from Mycobacterium ranae 
and from M. tuberculosis H37Rv.^^ 

15 E. Lederer, Angew. Chem. 72 372 (1960). 

i«L. G. Ginger and R. J. Anderson, /. Biol. Chem. 157 203 (1945) 
and preceding papers. 

^' Cecile Asselineau, Jean Asselineau, Ragnar Ryhage, Stina 
Stallberg-Stenhagen and Einar Stenhagen, Acta Chem. Scand. 13 
822 (1959). 

^^ Andrea V. Fowler, Merrill N. Camien and Max S. Dunn, J. Biol. 
Chem. 235 1386 (1960). 



Pfizer Handbook of Microbial Metabolites 666 

Lipides of Corynebacterium ovis have been studied/" 
as have the component fatty acids of Sporidesmium 
bakeri Syd. Hpides.-" 

The oil of wheat stem rust uredospores was found to 
contain a substantial quantity of an acid not previously 
reported from natural sources, cis-9,10-epoxyoctadecanoic 
acid, CisH;j40:t, colorless leaflets, m.p. 58.5-59.5°, cis- 
epoxide peak in the infra-red at 845 cm.'.-' 

The chemistry of naturally occurring 1,2-epoxides, in- 
cluding many microbial products, has been reviewed.''" 

Another new fatty acid, Ci^H.joO^, containing a cy- 
clopropane ring has been reported (in a preliminary com- 
munication) as occurring in Escherichia coli lipides." 

Bongkrekic acid, at a concentration of lO" molar, is a 
potent inhibitor of oxidative phosphorylation as carried 
out by mitochondrial enzymes in heart muscle tissue.-' 

The direct participation of protein-bound biotin in 
fatty acid biosynthesis has been confirmed.-* 

Both 9- and 10-hydroxystearic acids can replace oleic 
acid as growth factors for anaerobically grown yeast, 
which requires unsaturated acid, and these substances 
may be precursors of oleic acid in yeast. -"^ 

The role of vitamins in lipide metabolism has been re- 
viewed.-'"' 

Hydroxypyruvic acid has been isolated as the 2,4-dini- 
trophcnylhydrazone from Aspergillus nigcr. It may arise 
from 3-phosphoglyceric acid.'-' 

i^A. Diara and J. Pudles, Bull. soc. chim. biol. 41 481 (1959). 

2" L. Hartman, J. C. Hawke, Isobel M. Morice and T. B. Shorland, 
Biochem. J. 75 274 (1960). 

-' A. Tulloch, B. Craig and G. Ledingham, Can. J. Microbiol. 5 
485 (1959). 

-i» A. D. Cross, Quart. Revs. II 317-336 (1960). 

" Simone Dauchy and Jean Asselineau, Compt. rend. 250 2635 
(1960). 

-i W. Welling, J. A. Cohen and W. Berends, Biochem. Pharmacol. 
3 122 (1960). 

-' S. J. Wakil and D. M. Gibson, Biochim. et Biophys. Acta. 41 122 
(1960). 

'-'■'W. J. Lennarz and Konrad Bloch, ;. Biol. Chem. 235 PC 26 
(1960). 

-"Bacon F. Chow, Avi. J. Clinical Nutrition 8 321 (1960). 

-'Francis J. Behal, Arch. Biochem. and Biophijs. 88 110 (1960). 



667 Addendum 

The oxidative degradation of glycolic acid in £. coli 
takes the following course:-" 



-2H -f Acetyl CoA 

HOCH COOH • OHC -COOH • HOOC -CH— CH,— COOH 

— Acetyl COA | 

OH 

O O +H-0 

-2H I -CO, l| +CoA 

' HOOC -C CH —COOH > CH.CCOOH Acetyl CoA-^ 

-2H, -CO: 
Summation: OHC COOH r O: • 2CO2 h H,0 

A study has been made of the synthesis of cell mate- 
rials from acetate by Aspergillus niger^-^' and by Esche- 
richia coli.-''' Interrelationships of the tricarboxylic acid 
and glyoxylic acid cycles were discussed. 

Lactobacilli produce ^-hydroxy acids other than lactic. 
Two of these have been identified as a-hydroxy-D-isovaleric 
and D-isocaproic acids;-'' 

CH3 CH3 

\, \ 

CH— CH— COOH CH— CH3— CH— COOH 

/I /I 

CH3 OH CH3 OH 

D-lsovaleric Acid D-isocaproic Acid 

These are growth promoters for certain strains of lacto- 
bacilli. 

Penicilliiim atrovenetum, a /^-nitropropionic acid pro- 
ducer,'^'' was grown on C'^-labeled fj-alanine, on NaHC'^O., 
and on 4-C''*-D,L-aspartic acid.'' Since 96 percent of the 
label was in the 1 -position, apparently aspartic acid was 
incorporated as a unit. 

2^H. L. Kornberg and J. R. Sadler, Nature 185 153 ri960). 

2«' J. F. Collin and H. L. Kornberg, Biochem. J. 77 430 (1960). 

^s'-H. L. Kornberg, P. J. R. Phizackerly and J. R. Sadler, ibid. 77 
438 (1960). 

-■' Merrill N. Camien, Andree V. Fowler and Max S. Dunn, Arch. 
Biochem. and Biophys. S?> 408 ri959). 

■'^'H. Raistrick and A. Stossl, Biochem. J. 68 647 (1958). 

•'' A. J. Birch, B. J. McLoughlin, Herchel Smith and J. Winter, 
Chem. and Ind., 840 (1960). 



Pfizer Handbook of Microbial Metabolites 668 

A review of naturally occurring nitre compounds has 
been published.'*^ 

The fatty acids of B. alcaligenes faecalis, S. pullorum, 
B. fluorescens, S. typhi-murium and B. natta have been 
analyzed. ^^ Palmitic and unsaturated Cig-acids were the 
main components. Unsaturated Cig-acids were present 
to some extent, the unsaturated Cis- and Cjc-acids being 
largely oleic and palmitoleic. A saturated Cig-acid was 
abundant in the fat of B. natta. 

Two acids, 13-methyltetradecanoic, m.p. 52.5-53°, and 
15-methylhexadecanoic, m.p. 61.0-61.5°, were the main 
components of the fatty acid fraction of B. subtilis.^* 

A new monounsaturated, monohydroxy acid, diphthero- 
corynic, C53H104O3, has been reported produced by Coryne- 
bacterium diphtheriae .^^ Its relationship to related com- 
pounds has been discussed.^'' 

4. Tetronic Acids and Other Lactones and Lactams 

Tenuazonic acid ( 3-acetyl-5-sec-butyltetramic acid) 
has been biosynthesized incorporating 3.9 percent of the 
tracer from a medium containing CH^C^OONa.^^ Of the 
total incorporated radioactivity 96 percent was present in 
the C-2 and C-6 atoms. The remaining 4 percent was 
shared by C-4 and C-10, and this was explained on the 
basis of the manner of biosynthesis of isoleucine. 

4 3 6 7 

HO— C=C— CO— CH3 



10 9 8 1 

CH3— CH2— CH— CH C 

I 5'\ 



CH3 N 

11 H' 



\ 



32 M. Pailer, Fortschr. Chem. org. Naturstoffe 18 55-78 (1960). 

33Kunihiko Saito, /. Biochem. (Tokyo) 47 699 (1960). 

^^Idem., ibid. 47 710 (1960). 

3^ E. M. Gubarev and L. M. Pustovlova, Ukrain. Biokhim. Zhiir. 30 
569 (1958). 

3" Raoul Toubiana and Jean Asselineau, Compt. rend. 251 884 
(1960). 

3" C. E. Stickings and R. J. Townsend, Proc. Biochem. Soc, 36P 
(1960). 



669 Addendum 

It might be pointed out that, formally, some of the 
vulpinic acids are tetronic acids although we have not 
classified them as such. 

The chemistry of the tetronic acids has been reviewed.^® 

5. Carotenes and Carotenoids 

Another paper has been published on the incorporation 
of C^Mabeled compounds into carotenes by Neiirospora 
crassa.^'^ Mevalonic acid salts were the best of eight pre- 
cursors used, but less than 1 percent of the 2-C'* activity 
was incorporated into the carotene fraction. Phytoene, y- 
carotene and its isomers (fS- and ^-), phytofluene, neuro- 
sporene, spirOloxanthin and its isomers and lycopene 
were isolated. The presence of much phytoene, whose 
presence in the theoretical biosynthetic sequence has been 
questioned, was taken as an argument against formation 
of the carotenes by stepwise interconversions involving 
either hydrogenation or dehydrogenation and as an indi- 
cation, rather, of independent synthesis. 

The major carotenoids of some ascomycetes and basid- 
iomycetes have been identified. *° /^-Carotene was predom- 
inant in Epichloe typhina and Helotium citrinum. Crypto- 
xanthin was second in importance in Calocera viscosa. 
Neurosporene was the major carotenoid in dull yellow 
Cantharellus infundibiliformis with traces of lycopene 
present. The reverse was true in Cantharellus lutescens. 
No carotenoids, but instead pigments with quinone-Hke 
reactions, were detected in the grey Cantharellus cinereus 
and orange-red Giiepinius helvelloides. 

A red pigmented yeast isolated from root nodules of 
Lupinus luteus produced torulene, ^-carotene, y-carotene 
and torularhodin.*^ Diphenylamine inhibited production 
of y-carotene and torularhodin. 

Rhodotorula mucilaginosa contained, in decreasing or- 

38 L. J. Haynes and J. R. Plimmer, Quart. Revs. 14 292 (1960). 

33 Leo F. Krzeminski and F. W. Quackenbush, Arch. Biochem. and 
Biophijs. 88 287 (1960). 

*o Gilbert Turian, Arch. Mikrobiol. 36 139 (1960). 

" Gy. Schneider, B. Matkovics and J. Zsolt, Acta. Univ. Szegedien- 
sis. Acta. Phys. et Chem. 5 55 (1959). 



Pfizer Handbook of Microbial Metabolites 670 

der of quantity, torularhodin, torulene, y-carotene and f3- 
carotene, but no phytoene or phytofluene.*- Ultraviolet 
irradiation gave stable strains varying greatly from the 
parent both in quality and quantity of carotenoid content. 
One of many inhibitors tested, 2-hydroxybiphenyl, inhib- 
ited carotenogenesis without affecting culture growth. 
Doubt was expressed that the different carotenoids are 
biosynthetically mutually related. 

Oil of wheat rust (Puccinia gravtinis var. tritici) uredo- 
spores contained fj- and y-carotenes with minor amounts of 
phytoene, lycopene, a cis-/;j-carotene and a cis-carotene.*^ 

Mycoxanthin is the principal carotenoid of Mycobacte- 
rium battaglini.** Leprotene, a leprotene derivative, ^- 
carotene, a-carotene and an a-carotene monoepoxide prob- 
ably were present. 

A carotenoid pigment in Spirobacillus cienkowskii 
Metchnikoff, a pathogen of cladocera, resembled rhodo- 
violascin or a-bacteriopurpurin.*' Astacene and astaxan- 
thin also were thought to be present. 

Staphylococcus citreus contains the orange carotenoid. 
sarcinaxanthin, and the yellow sarcinene.**' Reference 
was made to two other uncharacterized carotenoids which 
have been isolated from natural sources, neoxanthin and 
corynexanthin.*' 

A new carotenoid has been isolated, which probably 
has the structure shown below. **^ 

175a Bacterioruberin a, C4oHr,,50o, mauve-violet needles, m.p. 182° 
(vac), U.V. 369, 385, 461, 494, 528 m^ in petroleum 
ether. 

"Jean Villoutreix, Biochim. et Biophys. Acta. 40 434, 442 (1960). 

*-^ F. Hougen.'B. Craig and G. Ledingham, Can. J. Microbiol. 4 
521 (1958). 

44Aldo Gaudiano, Rend. ist. super, sanita 22 769 (1959). (Chem. 
Abstr. 54 13253a) 

45 J. Green, Nature 183 56 (1959). 

**' Tatsuo Ohta, Toshio Miyazaki and Teruo Ninomiya, Chem. & 
Pharm. Bull. (Tokyo) 7 254 (1959). 

*" W. Hodgklss, J. Liston, T. W. Goodwin and Malini Jamikorn, 
J. Gen. Microbiol. 11 438 (1954). 

*^ Synnove Llaaen Jensen, Acta Chem. Scand. 14 950 (1960). 



Gyi Addendum 



) 



/ 111 

Halobacterium sp. 

A mutant of Staphijlococcus aureus unable to produce 
bright pigments incorporated the label of 2-c^*-mevalonic 
acid into phytoene, which it accumulated.*''" 

The biosynthesis and function of the carotenoid pig- 
ments have been reviewed.*-' Also a review of cis, trans- 
isomeric carotenoid pigments has been published.'^'' 

6. Polyenes and Polyynes, Excluding Polyene Macrolides 

In a review of polyacetylenes"^^ a number of substances 
not included in our list were mentioned without refer- 
ences or physical properties. These are reproduced here: 

193a Octa-2,6-dien-4-yn-l,8-dioic Acid, C8H6O4. 

HOOC— CH=CH— C = C— CH=CH— COOH 

Polyporus anthracophilus 

194a Non-2-en-4,6,8-triynoic Acid, CgH402. 

HC=C— C=C— C=C— C=C— CH=CH— COOH 

Psilocybe sarcocephala 
195a Non-2-frans-oxido-4,6,8-triynol (Biformin?), CgHgOo. 

HC=C— C=C— C^C— CH— CH -CH2OH 

trans 

Coprinus quadrifidis {Polyporus biformis?) 

*®° Ginzaburo Suzue, Biochim. et Biophys. Acta 45 616 (1960). 
«T. W. Goodwin, Advances in Enzymol. 21 295-361 (1959). 
^" L. Zechmeister, Fortschr. Chem. org. Naturstoffe 18 (1960). 
51 E. R. H. Jones, Proc. Chem. Soc, 199-211 (1960). 



Pfizer Handbook of Microbial Metabolites 672 

195b Non-4-cw-en-6,8-diynoic Acid, C9Hg02. 

HC^C— C=C— CH=CH— CH2— CH2— COOH 

CIS 

Drosophila subatrata 

198a Dec-2-<rans-en-4,6,8-triynoic Acid, C10H6O2. 

CH3— C^C— C = C— C=C— CH=CH— COOH 

frans 

Pleurotus ulmarius, Tricholoma paneolum 

201a Dec-2-frans-en-4,6,8-triynol, CioHgO. 

frans 
CH3— C=C— C=C— C=C— CH=CH— CH2OH 

Pleurotus ulmarius 
200a Deca-4,6,8-triyn-l,10-dioic Acid, C10H6O4. 

HOOC— C=C— C=C— C^C— CH2— CH2— COOH 
Merulius lachrymans 
219a Tetradec-5-cis-en-8,10,12-triyn-l,14-dioic Acid, C14H10O4. 

HOOC— C^C— C=C— C=C— CH— CH=CH— CH,— CHo— CH2— COOH 

CIS 

Poria sinuosa 

Four other polyacetylenes have been reported, complete 
with physical properties : ^^ 

195c Drosophilin E (cis-Non-4-en-6,8-diynoic Acid), CgHgOa, light- 
sensitive prisms, m.p. 35°, U.V. 279.5, 264, 250, 238, 227, 
210 m/x. 

CIS 

HC=C— C^C— CH=CH— CH2— CHo— COOH 

Drosophila subatrata 

209a Drosophilin C (cis-Undec-3-en-5,7,10-triynoic Acid), CnHgOs, 
colorless needles, slowly yellowing in light at 20°, m.p. 
97.5-99°, U.V. 280.5, 264.5, 250.5, 238, 226.5, 210.5 m,x. 

HC=C— CH2— C^C— C=C— CH=CH— CH2— COOH 

Drosophila subatrata 

52 E. R. H. Jones, P. R. Leeming and W. A. Remers, /. Chem. Soc, 
2257 (1960). 



673 Addendum 

209b Drosophilin D (cis-Undeca-3,9,10-trien-5,7-diynoic Acid), 
C,,HsOo, colorless plates, m.p. 21-28°, U.V. 303.5, 290.5, 
274.5, 259, 217 m^. 

H,C=C=CH— C=C— C^C— CH--CH— CH, -COOH 
Drosophila subatrata 

219b Compound 3040 (Dimethyl trans-Undeca-2-en-4,6-diyn-l,ll- 
dioate), C,.,Hi404, colorless crystals, m.p. 15-16°, U.V. 
304, 286, 270, 255, 222.5, 215 m/x. 

CH3OOC— CH2— CH2— CH2— C=C— C^C— CH=CH— COOCH3 

Drosophila subatrata 

An Italian review on the chemical aspects of the basid- 
iomycete antibiotics has been published. ''■^ 

7. Macrocyclic Lactones (Macrolides) 

A new tetraene antibiotic has been reported.^* 

233a Unamycin A, white needles, m.p. (dec.) 148-150°, [ajn^^ —92° 
(c 1.0 in 80% methanol-water), U.V. 290, 304, 319 m/x 
in methanol. 

An acidic tetraene. Negative FeCl^, Million, Fehling, 
Tollens tests. Positive Molisch, KMn04 and Br^ tests. 
Pink Schiff test. 

A second substance resembling toyocamycin was iso- 
lated : 

1288a Unamycin B, white needles, m.p. 236-238° (dec), [ajn^^ -43° 
(c 1.0 in acid methanol), N. E. 310. 

C 46.4, H 4.46, N 22.25. Gives essentially the same color 
tests as unamycin A. 

The unamycins were produced by Streptomyces fungi- 
cidicus. 

A heptaene which may be new has been reported." 

256a Grubilin green-yellow, amorphous. 

A non-toxic heptaene produced by Streptomyces BA-27, 

s^Marcella Magliola, Annali di Chimica 50 455-490 (1960). 

^* Masayuke Matsuoka and Hamao Umezawa, /. Antibiotics 
(Japan) 13A 114 (1960). 

" J. Uri, I. Szilagyi and I. Bekesi, Symposium on Antibiotics, 
Prague, 1959. 



Pfizer Handbook of Microbial Metabolites 674 

and differing from amphotericin B, ascosin, airreofacin, 
AYF, candicidin, candidin, candimycin, PA 150 and tri- 
chomycin. 

Antimycoin has been separated into A and B compo- 
nents.'^'' Mevalonic acid stimulated production of these 
substances by Streptoinyces aureus. Of nine other 
polyene producers tested, Streptoinyces viridoflavus pro- 
duction of candidin and Streptoinyces strain 3832 produc- 
tion of a pentaene (antibiotic S-8) of the eurocidin type 
were stimulated by mevalonic acid addition. 

The mechanism of nystatin action on Candida albicans 
has been studied.''' Respiration was accelerated and glu- 
cose uptake diminished, apparently by alteration of cell 
permeability. 

A dissertation has been published (not yet received) 
entitled Beitrag zur Kentitnis des Candicidins D, G. 
Demuth, Math.-Naturw. Fakultat der Univ. Gottingen, 
1959. 

Some generalizations can be made now concerning the 
structures of polyene macrolides.* Tetraenes and hep- 
taenes generally seem to contain nitrogen, while pentaenes 
and hexaenes do not. Moldicidin and PA- 153 are excep- 
tions since they are nitrogen-containing pentaenes. All 
tetraenes except PA-166 contain mycosamine. PA-166 
contains an amino sugar (not a deoxy type) other than 
mycosamine. Pentaenes are neutral, containing neither 
amino sugars nor free carboxyl groups. 

Heptaenes have been found so far to contain four dif- 
ferent nitrogen-containing moieties. Two of these are 
the amino sugars previously mentioned. The other two 
are the aromatic amines, p-aminoacetophenone and p- 
aminophenylacetone, which are released by alkahne hy- 
drolysis. " 

s« Robert Samuel Safferman, Dissertation Abstr. 20 4264 (1960). 

'"' J. W. Hai-man and J. G. Masterson, Irish J. Med. Sci. -$78 249 
(1957). 

* Most of the information below on the polyene macrolides was 
taken from a seminar given by Dr. Edward Borowsky, Visiting Pro- 
fessor at the Institute for Microbiology at Rutgers University from 
Gdansk, Poland, in August 1960 and will be published. 



675 Addendum 

O O 

H,N— / \— C— CH3 H,N— / \— CH,— C— CH3 
p-Aminoacetophenone p-Aminophenylacetone 

Amphotericin B and candidin are examples of heptaenes 
containing mycosamine. Candicidin, trichomycin and 
PA-150 contain p-aminoacetophenone. 

Hydrocandidin has yielded an oxidation fragment iden- 
tified as: 

HOOC— CH-(CH,)u— CH— CO— CH3 

I I 

CH3 CH3 

Some studies on the biosynthesis of this heptaene show 
no incorporation of labeled inevalonic acid, propionic acid 
or methionine. It seems to be derived from acetate. 

The pentaene, moldicidin A, C4;{H220i9N was omitted. 

Moldicidin B is identical with pentamycin."" Can- 
dicidin is identical with ascosin. The main component 
of the PA-150 complex is identical with one compo- 
nent of the candidin complex. Several substances listed 
in the unclassified section are actually known to be 
polyene macrolides. These include: 1072-aliomycin 
(pentaene), 1067-akitamycin (tetraene), 1095-antibiotic 
from Streptomyces fimgicidicus (tetraene), 1096-antibi- 
otic from S. griseus (heptaene), 1097-antibiotic 26/1 
(heptaene), 1294-substance 1404 (hexaene). 

A new heptaene, perimycin (aminomycin), probably 
C47H-.-,Oi4N2 and incorporating a p-aminophenyl group, 
has been reported. ^"^ Another heptaene, antibiotic 2814H, 
is produced together with a pentaene, antibiotic 2814P, 
netropsin and aureothin by Streptomyces lA 2814 resem- 
bling S. netropsis.-'-' Analytical and optical data were re- 
ported on each. 

•"" Hiroshi Ogawa, Teiichiro Ito, Shigeharu Inoue and Motohiro 
Nishio, /. Antibiotics (Japan) 13A 353 (1960). 

■''''Edward Borowsky et al.. Abstracts 1960 Conference on Anti- 
microbial Agents, Washington, D. C, October 26-28, 1960. 

'''■' Heinz Thrum and I-dschang Dcho, N atiirwissenschaften 20 474 
(1960). 



Pfizer Handbook of Microbial Metabolites 676 

The complete structures of the tetraenes, lagosin and 
filipin have been reported to be : ^° 

CsHu— CH— OH OH OH R=OH=lagosin, C35H58O12 

I I I R=H=filipin, CssHssOu 

O— CO— CH (CH— CH2)5— CH— CH— R 

I 1 

CH3— CH— CH— (CH=CH)4— CH=C CH— OH 

I I 

OH CH3 

Humidin (Cj2H2o04)n, colorless plates, m.p. 145-146° 
(dec), [aln'* -6° (c 1.0 in ethanol), mol. wt. 550 ± 50, 
823 ± 10, is an antifungal antibiotic isolated from the 
myceUum of Streptomyces humidus, which also produces 
dihydrostreptomycin.^^ It was not clear from the abstract 
whether or not this substance was of the polyene mac- 
rolide type. 

Some aspects of the mode of action of polyene anti- 
fungal antibiotics have been reviewed."^ 

A nitrogen-containing antifungal polyene antibiotic, 
capacidin, produced by a streptomycete has been iso- 
lated.®^- ®* The substance is levorotatory, has reducing 
properties, is a primary or secondary alcohol and shows 
ultraviolet absorption peaks at 318, 332, 350 m^^. 

A general review of the polyene antifungal antibiotics 
has been published. '^^ 

Two new antibiotics have been reported, one of them, 
at least, apparently a macrolide.*^® 

^^M. L. Dhar, V. Thaller and M. C. Whiting, Proc. Chem. Soc, 
310 (1960). 

**i Koichi Nakazawa, Motoo Shibata, Hiroichi Yamamoto, Toshihiko 
Kanzaki, Eiji Higashide, Akira Miyake and Satoshi Horii, Nippon 
Nogei Kagaku Kaishi 32 713 (1958). (Chem. Abstr. 54 22843g) 

^^ E. Drouhet, L. Hirth and G. Lebeurier, Annals N. Y. Acad. Sci. 
89 134-155 (1960). 

83 Rachel Brown and Elizabeth Hazen, N. Y. State Dept. Health, 
Ann. Rept. Div. Labs, and Research 50-52 (1959). (Chem Abstr. 
54 22824h) 

^Idem., Antibiotics and Chemotherapy 10 702 (1960). 

«5L. C. Vining, Hindu Antibiotics Bulletin 3 37-55 (1960). 

8" E. Gaumann, R. Hiitter, W. Keller-Schierlein, L. Neipp, V. Prelog 
and H. Zahner, Helv. Chim. Acta 43 601 (1960). 



677 Addendum 

265a Lankamycin, C3cHe20i4, colorless crystals, m.p. 147-150° and 
at 181-182°, "[a]ir" -94° (c 1.23 in ethanol). U.V. 289 
ni/x. 

Typical erythromycin color tests were obtained. It is 
notable that this macrolide contains no amino sugar. 

A second, unclassified antibiotic was isolated from the 
same culture (Streptomyces violaceoniger (Waksman et 
Curtis) (Waksman et Henrici). 

n64a Lankacidin, C4,5H,;,-Oi6N2, pale yellow microcrystalline powder, 
m.p. 165-168°, [aW -161° (c 0.967 in ethanol), U.V. 
227 m^ (log 2.95). 

Contained no N — CH3 or — OCH3 groups. 

It is interesting that spiramycin contains three sugars. *^^ 

A paper on the mode of action of erythromycin"* reports 
that, when the antibiotic was added to growing cells of 
E. coli, synthesis of protein (but not RNA or DNA) was 
inhibited, as was adaptive formation of ^-galactosidase. 
Lactose was the substrate. Oxygen uptake of resting cells 
was inhibited in some organisms but not in others, but in 
no case did cytochrome oxidase appear to be affected. 

The wild strain of Streptomyces kitasatoensis Hata pro- 
duces leucomycin, a complex of six macrolide antibiotics, 
while a mutant produces only two of these, although total 
macrolide production was the same in each case.*"'^ "°- "^ 
Probable empirical formulas of the members of the com- 
plex are shown below : 

leucomycin formula melting point 

Ai C46HS1 O17N 

Ai C65H111O22N 

Bi C36H59O13N 214.5-216.5° 

B2 CssHesOieN 214-216° 

B3 C34H63 0,3N 216-217° 

B4 C38H59O16N 221-223.8° 

^'' Raymond Paul and Serge Tchelitcheff, Bull. Soc. chim. France, 
150 (1960). 

^^ Hiroshi Nakagawa, Osaka Daigaku Igaku Zasshi II 3451 (1959). 
(Chem. Abstr. 54 11154a) 

'^^ J. Abe, Y. Suzuki, T. Watanabe and K. Satake, Nippon Kagaku 
Zasshi 31 969 (1960). 

■OT. Watanabe et al. Bull. Chem. Soc. Japan 33 1100 (1960). 

^^ Tetsuo Watanabe, Hisao Nishida, Jinnosuke Abe and Kazuo 
Satake, ibid. 33 1104 (1960). 



Pfizer Handbook of Microbial Metabolites 678 

Methymycin has been found to be biosynthesized prin- 
cipally from propionate, although one mole of acetate 
may be incorporated. '- 

8. Alicyclic Compounds Other Than Terpenoids and 
Steroids 

An investigation of the biosynthesis of palitantin shows 
that it is acetate-derived, and that neither shikimic acid 
nor mevalonic acid are involved.'^ 

Several compounds have been isolated which may be 
related to cycloheximide : 

302a Niromycin B, C14H21O4N (suggested), white, hygroscopic crys- 
tals, m.p. 47-67°. 

A neutral substance produced by Streptomyces albusJ* 

302b Niromycin A, white hygroscopic, amorphous powder, m.p. 98- 
105°. 

Positive 2,4-dinitrophenylhydrazine and Tollens tests. 
Negative ninhydrin, FeCla, Fehlings, Benedicts, Molisch, 
biuret, KMn04 tests. 

The effect of cycloheximide on the metabolism and 
growth of Saccharomyces pastoriamis has been studied."'^ 

Some substances related to sarkomycin and produced 
by the same organism were overlooked.''' These were: 

301a Sarkomycin Eo, C10H14O4, m.p. 179°. 

301c Sarkomycin Ep Ci4HisOt, m.p. 169°. 

sold Sarkomycin So, Ci4Hi,OoS, m.p. 183°, [a]n +136°. 

COOH COOH 





CH,— S— CH. 

o o 

''-A. J. Birch, E. Pride, R. W. Rickards, P. J. Thomson, J. D. 
Dutcher, D. Perlman and C. Djerassi, Chem. and hid., 1245 (1960). 

■■^ P. Chaplen and R. Thomas, Biochem. J. 77 91 (1960). 

'* Teisuke Osato, Yutaka Morikubo and Hamao Umezawa, J. Anti- 
biotics (Japan) 13A 110 (1960). 

"5 Bradner Wood Coursen, Dissertation Abstr. 21 (1960). 

'« Sueo Tatsuoka et al., J. Antibiotics (Japan) 9B 104 (1956). 




679 Addendum 

30le Sarkomycin S^, CnH,sO,;So, m.p. 161°, [a],," +145°. 
COOH COOH 

/ y-CHj— S— S— CH 

\ / 

o o 

30if Sarkomycin S;;, m.p. 148°. 

C 50.39, H 5.31, S 15.75. 

9. Terpenoids and Steroids 

A new trichothecin-like antibiotic has been isolated 
from a basidiomycete." 

The oil of wheat stem rust uredospores contains A'- 
ergostenol (fungisterol).'*-^ 

Another steroidal metabolite of Poria cocos has been 
isolated and characterized. It is:" 

354a Pachymic Acid ( 3/3-0-Acetylpolyporenic Acid B), CasH-.^O-,, col- 
' orless crystals, m.p. 296-299°, [a]D"' 17.7° (c 0.566 in 
pyridine ) . 

HOOC 




CH3COO 



Cholesterol biosynthesis is inhibited by farnesoic acid 
and its analogues.'^ 

The conversion of mevalonate to a mixture of farnesol 
and nerolidol (probably as their pyrophosphates) by a 

" Ervln Glaz, Eszter Scheiber, J. Gyimesi, I. Horvath, Katalin 
Steczek, A. Szentirmai and G. Bohus, Nature 184 Suppl. No. 12, 908 
(1959). 

'^ Shoji Shibata, Shinsaku Natorl, Ko Fujita, Isao Kitagawa and 
Kazue Watanabe, Chem. & Pharm. Bull. (Tokyo) 6 608 (1958). 

'^G. Popjak, Rita H. Cornforth and K. Clifford, Lancet, 1270 
(1960). 



Pfizer Handbook of Microbial Metabolites 680 

liver enzyme preparation has suggested that 1 mole of 
each is involved in the biosynthesis of squalene.®° The 
condensation of these two substances would then be anal- 
ogous to that of isopentenylpyrophosphate with 3,3-di- 
methylaUyl pyrophosphate (or geranyl pyrophosphate). 

CH=-OPO.H, 




Nerolidyl Pyrophosphate 



Squalene 

The significant points of the chemical mechanism of 
squalene biosynthesis were summarized as follows: 
(a) The process is not a concerted reaction, but proceeds 
in steps with well-defined stable intermediates, (b) Dur- 
ing isomerization of isopentenylpyrophosphate there is an 
uptake of one proton in the terminal methylene group, 
and this proton appears finally in one of the terminal 
methyl groups at each end of squalene, which means the 
entry into each molecule of squalene of two protons not 
contained originally in mevalonic acid, (c) There are no 
reductive steps involved in the synthesis of geranyl or 
farnesyl pyrophosphates, (d) Farnesyl pyrophosphate 
and the nerolidyl derivative are the two sesquiterpenoids 
condensing to the symmetrical dihydroterpene, squalene, 
a stable intermediate being dihydrosqualene. (e) During 
stabilization of the condensation product of the farnesyl 
and nerolidyl derivatives, elimination of two protons, orig- 
inally attached to C-5 of mevalonate occurs, (f) The 
final step is a reduction, introducing into squalene two 
further hydrogen atoms not contained originally in 
mevalonic acid. 

80 J. W. Cornforth and G. W. Popjak, Tetrahedron Letters No. 19 
29 (1959). 



68 1 Addendum 

10. Tropolone Acids 

More data have been published on the structure of 
heliomycin (entry 1173). It is acidic (pK 5.8), forms a 
diacetate and may contain a benzotropolone ring system. 
Empirical formulas Ci9Hi4.i(505 or CoaHjH joOe have been 
suggested.'*^ 

11. Phenolic Substances 

p-Hydroxybenzoic acid, found earlier in Penicillium 
patuhim has been isolated also from Penicilliiim griseo- 
fiilviim.^- Isolated from the same culture were: 

379a »j-Hydroxybenzoic Acid, CyHyO;}, m.p. 201° 

and 
379b Salicylic Acid ( o-Hydroxybenzoic Acid), m.p. 159°. 

The same mold produces homogentisic acid, a metabo- 
I lite also found in some of the higher fungl.®^ 

391a Homogentisic Acid, C8H8O4, m.p. 152-154°. 

COOH 
OH 




p-Hydroxyphenylpyruvic acid and tyrosine were identi- 
fied in the culture, and occasionally 1,4-hydroquinone 
was present. 

The production of galHc acid by Phycomyces blakeslee- 
anus (sporangiophores) has been confirmed.^* It was 
suggested that this substance may be the primary photo- 
sensitive pigment involved in the strong negative photo- 
tropic response to ultraviolet light which such organs 
show. 
*i Z. V. Pushkareva, N. M. Voronina, S. I. Omerchenko, L. B. 
Radina and Yu. N. Sheinker, /. Gen. Chem. (USSR) 29 3469 (Eng- 
lish translation) (1960). 

^- P. Simonart, A. Wiaux and H. Verachtert, Bull. soc. chim. biol. 
41 537, 541 (1959). 

^^ Paul Simonart, Anselme Wiaux and Hubert Verachtert, Zentrl. 
Bakteriol. Parasitenk. Abt. II 113 209 (1960). 
*'* David S. Dennison, Nature 184 2036 (1960). 



Pfizer Handbook of Microbial Metabolites 682 

C^*-Labeled orsellinic acid has been prepared by using 
Chaetomium cochliodes as the producer. Orsellinic acid 
was known to be a metabolite of Penicillium barnense, 
which also produces penicillic acid. When Penicillium 
barnense was grown in the presence of the labeled orsel- 
linic acid, it could be shown that orsellinic acid was a 
precursor of penicillic acid in this organism. ^^ The sites 
of labeling and actual modes of cleavage are shown. 

HO O CH2 CH3 
y C C 




COOH 



HC C 

V/x * 

h o + CO2 



0CH3 

Orsellinic Acid Penicillic Acid 

It appears that orsellinic acid is an intermediate in the 
biogenesis of the xanthone ravenehn.*'' It has been sug- 
gested as an intermediate in the biosynthesis of several 
other types of compounds, e.g., lichen substances, fungal 
anthraquinones and alternariol. 

An uncharacterized substance has been isolated from 
Curvularia lunata.^^ 

417a Substance from Curvularia lunata, Cj 411x^0.5, colorless solid, 
m.p. 195°. 

Apparently phenohc. Mannitol was isolated from the 
same culture. 

Curvularin, also produced by Curvalaria lunata, is pro- 
duced by Penicillium steckii as well.** 

A new depsidone has been isolated from an Australian 
Uchen and characterized as norlobaridone : *'* 

85 Klaus Mosbach, Acta. Chem. Scand. 14 457 (1960). 

86 Private communication from Herchel Smith. 

^~ T. Krishna Murty and S. Sankara Subramanian, Indian J. 
Pharmacij 20 72 (1958). 

»* D. Fennell, K. B. Raper and F. H. Stodola, Chem. and Ind., 1382 
(1959). 

*" G. P. Briner, G. E. Gream and N. V. Riggs, Australian J. Chem. 
13 275 (1960). 



683 Addendum 

471a Norlobaridone, C.-jHonOe, colorless crystals, m.p. 188-190°. 

CH3CH,CH,CH:CO ^^^ 

COO OH 




HO O I 

CH,CH.CH,CHoCH3 

A yield of 2.2% was obtained from Pannelia conspersa. 
The structure of nidulin (and thus of nornidulin) has 
been completed.^''' It is: 

^^' COO ^^' OCH3 




The chemistry of the uncommon 1-methyIpropenyl sub- 
stituent is greatly modified by the neighboring chlorine 
atom. 

12. Quinones and Related Compounds 

a. BENZOQUINONES 

The growth of a mycobacterium was stimulated by 
coenzyme Qio which suggests a possible role in energy 
metabolism.*^" 

b. NAPHTHOQUINONES 

A variety of bacteria (Bacillus cereus, B. subtilis, Pro- 
teus vulgaris, Sarcina flava. Staphylococcus aureus, My- 
cobacterium phlei, Pseudomonas spp., Azotobacter vine- 
landii, Nocardia sp.) were examined for vitamin K 
content.^^ Three types were identified. Vitamin K^ was 

*»" F. M. Dean, D. S. Deorha, A. D. T. Erni, D. W. Hughes and 
John C. Roberts, /. Chem. Soc, 4829 (1960). 

•"' James O. Norman and Robert P. Williams, Biochem. and 
Biophys. Res. Comms. 2 372 (1960). 

»i Bodil Kruse Jacobsen and Hendrik Dam, Biochim. et Biophys. 
Acta 40 211 (1960). 



Pfizer Handbook of Microbial Metabolites 684 

isolated from Bacillus cereus and vitamin K^ or a related 
substance from Mycobacterium phlei. 

A lipide cofactor, perhaps a K vitamin or a tocopherol, 
has been implicated in the conversion of L-gulonolactone 
into L-ascorbic acid.^^ 



C. ANTHRAQUINONES 

In 1955 three substances were isolated from a yellow 
sterile mold and were called flavomycelin, rhodomycelin 
and purpurmycelin.^^ Rhodomycelin is identical with 
islandicin and flavomycelin vvdth luteoskyrin. Acetone 
solutions of luteoskyrin turn purple on exposure to light, 
and purpurmycelin was found to be identical with this 
irradiation product.^* 

The biosynthesis of the pigments of Penicillium islan- 
dicum has been studied. ^^ The acetate origin of islandi- 
cin, skyrin, rubroskyrin (luteoskyrin) and iridoskyrin 
was established. The results of labeling experiments led 
to the conclusion that, despite the close structural rela- 
tionship, these pigments are not interconvertible in vivo, 
but seem to be derived from a common pre-aromatic 
stage. Also mutations fail to block formation of any sin- 
gle pigment. Biogenesis, it was suggested, must not take 
place by stepwise formations of defined intermediates 
such as benzene derivatives, but should be dependent 
throughout on participation of activated acetate. 

An acidic substance related to herqueinone has been 
isolated.^'' 

A review of quinones as metabolic products of micro- 
organisms has been published.''' 

There Jiave been two recent pubUcations on the struc- 



92 I. B. Chatterjee, N. C. Kar, N. C. Ghosh and B. C. Guha, Arch. 
Biochem. and Biophys. 86 154 (1960). 

93 H. Nishikawa, Tohoku J. Agr. Res. 5 285 (1955). 

9* S. Shibata, I. Kitagawa and N. Nishikawa, Pharm. Bull. (Tokyo) 
5 383 (1957). 

95 Sten Gatenbeck, Acta Chem. Scand. 14 102, 230, 296 (1960). 

96 K. S. Gopalkrishnan and N. Narasimhachari, "Antibiotics," 
Council of Scientific and Industrial Research, New Delhi, 1958, pp. 
176-179. 

9^ J. H. Birkinshaw, Planta Med. 7 367 (1959). 



685 



Addendum 



ture of thelephoric acid (entry 493).^^ "^ The second 
publication cited reported the synthesis of thelephoric 
acid and seems to establish the structure definitely as: 

O 



HO 



HO 




OH 



OH 



Oosporein (chaetomidin) (entry 487) is reported to be 
identical with isooosporein (entry 488).^"" 

A reinvestigation of quinones produced by Phoma ter- 
restris Hansen identified cynodontin and a small amount 
of another anthraquinone, but found no phomazarin (en- 
try 556).'"! 

13. Tetracycline, Analogues and Related Substances 

The aglycone, aklavinone, of the antibiotic aklavin has 
been found to differ from rutilantinone ( e-pyrromyci- 
none) only by lacking one hydroxyl group/"^ 



OH OH 




Aklavinone C22H20O8 



OH O 

COOCH3 

Rutilantinone C22H20O9 



A biogenesis was postulated in the following sense: 
0000 




^^ K. Aghoramurthy, K. G. Sarma and T. R. Seshadri, Tetrahedron 
Letters No. 16 4 (1960). 

9»J. Gripenberg, Tetrahedron 10 135 (1960). 

i°o J. Smith and R. H. Thomson, ibid. 10 148 (1960). 

"ID. E. Wright and K. Schofield, Nature 188 233 (1960). 

102 J. J. Gordon, L. M. Jackman, W. D. QUis and I. O. Sutherland, 
Tetrahedron Letters No. 8 28 (1960). 



Pfizer Handbook of Microbial Metabolites 686 

A more recent publication indicates that nine acetate 
units are incorporated into the rutilanlinone molecule, 
but that the three starred atoms are from propionate. ^°^ 
Methionine would not, then, be involved in the side-chain 
synthesis. 

Two investigations have been made on the chlorina- 
tion mechanism of Streptomyces aureofaciens in the pro- 
duction of aureomycin.^"^ "^ The authors of the first 
reference concluded that incorporation of the chlorine 
atom does not take place on the finished tetracycline 
molecule, but at an earlier stage of biosynthesis. Wang's 
results lead to the same conclusion. 

The influence of specific enzyme poisons on the produc- 
tion of oxy tetracycline has been studied. ^•'^ Iron-contain- 
ing oxidases and flavine oxidases participated in the 
biosynthesis of oxy tetracycline. Phenoloxidase inhibi- 
tors, on the other hand, stimulated production. 

There is httle agreement on the mode of action of the 
tetracycline antibiotics, and it may be that they act in a 
variety of ways. Inhibition of RNA and DNA synthesis 
and inhibition of enzymic conversion of uracil to thy- 
mine,^°^ binding by chelation of metal ions required by 
coenzymes^-"' and blocking of unspecified biosynthetic 
pathways^°^ have been mentioned. 

A discussion of the mechanisms of action of antibiotics 
in general has been published. ^^"^ 



103 w. D. Ollis, I. O. Sutherland, R. C. Codner, J. J. Gordon and 
G. A. Miller, Proc. Chem. Soc, 347 (1960). 

i°* J. Kollar and M. Jaral, Symposium on Antibiotics, Prague, 1959. 

105 E. Lin Wang, J. Antibiotics (Japan) 12A 31, 41, 50 (1959). 

loo V. Sevcik, V. Musilek and I. Komersova, Symposium on Anti- 
biotics, Prague, 1959. 

10' T. Balakrishna Rao, D. V. Temhane, D. V. Rege and A. Sreeni- 
vasan, "Antibiotics," Council of Scientific and Industrial Research, 
New Delhi, 1958, p. 212. 

108 E. U. Weinberg, Bacterial. Revs. 21 46 (1957). 

10^ J. F. Snell, Florence Z. Thanassi and Dorothy Ann Sypowicz, 
Antibiotics and Chemotherapy 8 57 (1958). 

110 S. G. Bradley and L. A. Jones, Annals N. Y. Acad. Sci. 89 123 
(1960). 



687 Addendum 

14. Aromatic Compounds Not Classified Elsewhere 

The cooccurrence of anisaldehyde and junipal in 
Dacdalca jutupcrina cultures has inspired the suggestion 
that both substances are derived from a common acety- 
lenic precursor.'"' "- An eai'lier report that Poliiporiis 
beuzoiuus produces considerable quantities of anisalde- 
hyde was not mentioned in our entry on that substance."^ 

15. Amines 

Although the ordinary source of the amine, carnitine, 
is mammalian muscle, a publication was overlooked in 
which it was isolated from the mold Neurospora crassa 
grown on a chemically defined medium."* 

653a L-Carnitine, C7H]-0;iN, extremely hygroscopic crystals, m.p. 
196-198°, [a],,'-" -23.5° (c 0.5 in water). 

® 

(CH3)3N— CH2— CH— CH2— COO ^ 

OH 

This amine would not replace choline in choline-less 
neurospora mutants. It was not found in E. coli. The 
role of carnitine in lipide metabolism has been re- 
viewed."^' ^^^ 

An amine related to muscarine has been isolated and 
characterized by synthesis.""' "* It is: 

658a ( + )-Muscaridine, C,,H..0._.NC1 (Chloroaurate), C.,H..AuCl40.N, 
m.p. 129-131°, UW -f20.5° ±0.5° (c 8.3 in water). 

"1 J. H. Birkinshaw and P. Chaplen, Biochem. J. 60 255 (1955). 

11- K. E. Schulte and N. Jantos, Arch. Pharm. 292 536 (1959). 

^^^ J. H. Birkinshaw, E. N. Morgan and W. P. K. Flndlay, Biochem. 
J. 50 509 (1952). 

"*G. Fraenkel, Biol. Bull. 104 359 (1953). 

"' G. Fraenkel and S. Freedman, Vitamins and Hormo7ies 15 74- 
115 (1957). 

""E. P. Adams, P. E. Ballance and A. E. Bender, Nature 185 612 
(1960). 

"■ F. Kogl, C. A. Salemink and P. L. Schuller, Rec. trav. chim. 79 
278 (1960). 

"'^C. A. Salemink and P. L. SchuUer, ibid. 79 485 (1960). 



Pfizer Handbook of Microbial Metabolites 688 

© CI e 

(CHalsN CH,— CHo— CHo— CH— CH— CH3 

I I 

OH OH 

Amanita muscaria 

A survey of 32 fungi and nine bacteria indicated that 
the production of choline sulfate is limited to the higher 
fungi. ^^^ All bacteria were negative as were phycomy- 
cetes. Of the ascomycetes, spharioles produced it, but 
endomycetales did not. Basidiomycetes and all fungi 
imperfecti examined (except Torula utilis) were pro- 
ducers. 

List has continued his investigations of the basic con- 
stituents of higher fungi. From Polyporus sulfureus 
were isolated the following non-volatile substances: ade- 
nine, hypoxanthine, arginine, histidine, lysine, choHne, 
histidine betaine, phenylethylamine, imidazolyl acetate, 
homarine, trigonelHne, y-butyrobetaine and an uncharac- 
terized hydrochloride, C9Hi6N2-2HCl.^-° 

The mushroom Coprinus atramentarius was studied. ^^^ 
A prior report that it produced tetraethylthiuram disul- 
fide could not be confirmed. Found, however, were iso- 
amylamine, phenylethylamine, adenine, hypoxanthine, 
urocanic acid, imidazolyacetic acid, imidazolylpropionic 
acid, imidazolylethanol, histidine, arginine, choline, ly- 
sine, guanidine, ergothioneine, hercynine, glycine, be- 
taine, tyramine, putrescine, cadaverine, S-aminovaleric 
acid, a-guanidinobutyric acid, two unidentified bases, 
glycine, threonine, glutamic acid, aspartic acid, alanine, 
proline, leucine, valine, isoleucine, citrulline, tyrosine 
and ornithine. 

A dissertation has been published on basic constituents 
and amino acids of the basidiomycete, Inocybe patoul- 
lardii Bres.^-- 

Found were methylamine, dimethylamine, ethylamine, 
n-propylamine, isoamylamine, /?-phenylethylamine, cho- 
line, cadaverine, putrescine, hypoxanthine, alanine, pro- 
lix T. Harada and B. Spender, /. Gen. Microbiol. 22 520 (1960). 
120 p. List and H. Menssen, Arch. Pharm. 292 260-271 (1959). 
^21 p. H. List and H. Reith, Arzneimittel-Forsch. 10 34-40 (1960). 
^2- H. Miiller, Dissertation, Naturw. Fakultat Univ. Wiirzburg, 
1959. 



689 Addendum 

line, tyrosine, valine, leucine, cysteine, aspartic acid, 
glutamic acid, histidine, imida/.ole-4-acctic acid, argi- 
ninc. ornithine and the incompletely characterized basic 
red pigment of the organism, C,,Ho„0,,N^,. This was yel- 
low in alkali, red in acid solutions and gave positive 
Bayer and Pauly diazo tests. 

A study of the biogenesis of spermidine (entry 642) in 
microorganisms has shown that the C4 moiety is derived 
from putrescine (or ornithine) while the C;; chain has its 
origin in methionine.'-'* 

Biochemical pathways in legume root nodule nitrogen 
fixation have been reviewed.'-* 

16. Amino Acids and Related Compounds 

The lysine, methionine and tryptophan contents of a 
number of yeasts have been surveyed.'" 

In a study of the interrelationships between folic acid 
and cobalamin in the synthesis of methionine by extracts 
of E. coll, it was concluded that serine is not on the route 
of biosynthesis of the methyl group of methionine.'-" 

Discussing the two modes of lysine synthesis by lower 
fungi, Vogel has pointed out that organisms of older evo- 
lutionary origin follow the bacterial route.'-' These in- 
clude eubacteria, pseudomonads and actinomycetes. As- 
comycetous and basidiomycetous fungi use the fungal 
pathway via a-aminoadipic acid. 

17. Polypeptides and Related Compounds 

The ostreogrycin (E-129) complex was isolated in 
1958'-^ and reported similar to streptogramin, staphylo- 
mycin (A-899), PA- 11 4 and mikamycin. 

E-129 A probably is identical with staphylomycin M and 



123 H. Tabor, S. M. Rosenthal and C. W. Tabor, /. Biol. Chem. 233 
907 (1958). 

124 F. J. Bergersen, Bacteriol. Revs. 24 246 (1960). 

'-'■ G. E. N. Nelson, R. F. Anderson, R. A. Rhodes, Margaret C. 
Shekleton and H. H. Hall, Appl. Microbiol. 8 179 (1960). 

126 R. L. Kislluk and D. D. Woods, Biochem. J. 75 467 (1960). 

12" H. J. Vogel, Biochim. et Biophijs. Acta 41 172 (1960). 

12^ S. Ball, B. Boothroyd, K. A. Lees, A. H. Raper and E. Lester 
Smith, Biochem, J. 68 24p (1958). 



Pfizer Handbook of Microbial Metabolites 690 

PA-114A. E-129B may be identical with PA-114B, but 
different from staphylomycin S : ^^^ 

770a Ostreogrycin B, (E-129B) C45H-4O9NS, colorless prisms from 
methanol with solvation, colorless needles from toluene, 
m.p. 266-268°, [aW -66.8° (c 0.5 in methanol). 




CH--CH^CH3 




N-Me-p- / ^O 

Dimethylamino- 



phenylalanine L-Pro 



N 



CH, 



-N C^^ \ / 



Streptomyces ostreogriseiis 

This structure differs from staphylomycin only by sub- 
stitution of p-dimethylamino-N-methylphenylalanine for 
N-methylphenylalanine. 

A similar structure has been proposed for mikamycin B, 
the only difference being a hydroxyl group in the /^-posi- 

'-" F. W. Eastwood, B. K. Snell and Alexander Todd, /. Chem. Soc, 
2286 (1960). 



691 Addendum 

tion of the pyridine moiety adjacent to the carbonyl 
group in the mikamycin. '-'•'" 

An antiviral polypeptide, ccphaloniycin, has been re- 
ported.'"' It contained leucine, alanine, valine, arginine, 
glutamic acid, aspartic acid, glycine, threonine, tyrosine, 
phenylalanine and three unidentified ninhydrin-positive 
substances. 

Some peptide sequences of colimycin have been deter- 
mined.'" It resembles polymyxin B, and the sequence 
L-Dia — L-Dia — D-Leu — L-Leu — L-Dia — L-Dia — L-Dia — l- 
Dia — L-Thr has been established (L-Dia = a, y-L-diamino- 
butyric acid). 

An antibiotic named colisan has been isolated from a 
bacillus.'^- '-'' 

Sporidesmolide I, a metabolic product of Sporidesmium 
bakeri Syd., colorless needles, m.p. 261-263°, [a],,'" 
—217° in chloroform (c 1.5) has the empirical formula 
C33H58OSN4 and the structure : "^^ 



CH3 CH3 

\ / 

CH 





CH3— CH 






\ 






CHo 







\ 


II 




CH- 


— c 




X^D 






NH 






v/ 




CH, 

\ 


C 

„ / 

"—CM 




C 




/ 






CH, 


NH 

\ 

CH 






CH3— CH ^ 

] 


— c- 

II 




1 
CH3 


II 





\^° 



CH3 



/ \ 



C' CHj 



CH; 



-CH 

\ 
CH. 

\ 
CH 

/ \ 
CH3 CH3 

129a xiyoshe Watanabe, Hiroshi Yonehara, Hamao Umezawa and 
Yusuke Sumiki, /. Antibiotics (Japan) 13A 293 (1960). 

""Akihiro Matsumae, /. Antibiotics (Japan) 13A 143 (1960). 

"^ Michel Dautrevaux and Gerard Biserte, Compt. rend. soc. biol. 
153 1346 (1959). 

132 R. Reitler and J. Boxer, Nature 158 26 (1946). 

133 R. Reitler and A. Berner, to be published. 

"3" D. W. Russell, Biochim. et Biophijs. Acta 45 411 (1960). 



Pfizer Handbook of Microbial Metabolites 692 

making it a new member of the depsipeptide or peptolide 
class. This is the first report of L-a-hydroxyisovaleric acid 
as a natural product. 

A wilt toxin, culmomarasmin, which was 200 times as 
active as fusaric acid or lycomarasmine, has been isolated 
from Fusarium culmorura}^^ It is a crystalline polypep- 
tide, m.p. 215-218° (dec), stable below 0°. It is 
ninhydrin-negative and has the analysis: C 45.31, H 7.08, 
O 27.56, N 10.36, S 4.76, CI 4.19, C— CH3 3.17, — OCH3 
1.08. It also contains iron (1.39% inorganic residue). 
The amino acids are cystine, leucine, serine, aspartic 
acid, glutamic acid, alanine, valine, affo-isoleucine, 
proline, glycine, threonine and ammonia. 

Two dissertations on wilt toxins have been pub- 
lished.i^^- ^^^ 

The antibacterial activities of acyclic decapeptide ana- 
logues of gramicidin S have been measured."^ The mode 
of action of the acyclic compounds differs from that of 
the cycHc ones. While gramicidin S causes immediate 
bacteriostasis, the acyclic analogues are effective only 
after several cell divisions. The most active analogue 
was Yio as active as gramicidin S against E. coli and Mo 
as active against Staphylococcus aureus. 

The mushroom toxin, phalloidin, has been reported to 
act by inhibition of oxidative phosphorylation.^^* A more 
recent study claims that it acts, rather, by interference 
with protein synthesis."" 

The neuromuscular blocking properties of various poly- 
peptide antibiotics have been investigated.""" 

A yellow pigment has been isolated from E. coli.^*^ 

"*J. Kiss, Chimia 14 174 (1960). 

1^5 Hans Gempeler, tjber welkakdve Inhaltsstoffe von Endopathia 
parasitica (Murr.) und von Fusarium martii. Dissertation, Eidgenos- 
sische Technische Hochschule, 1959. 

136 Fritz Kugler, tjher welkdktive Inhaltsstoffe von Endopathia 
parasitica (Murr.) und von Fusarium solani (Mart.) v. Martii, Dis- 
sertation, Eidgenossische Technische Hochschule, 1959. 

137 B. F. Erlanger and L. Goode, Science 131 669 (1960). 
"SBenno Hess, Biochem. Z. 328 325 (1956). 

"9 A. von der Decken, H. Low and T. Hultin, ibid. 332 503 (1960). 

i39« R. H. Adamson, F. N. Marshall and J. P. Long, Proc. Soc. 
Exptl. Biol, and Med. 105 494 (1960). 

1*" K. Ishii and M. Sevag, Arch. Biochem. and Biophys. 77 41 
(1958). 



693 Addendum 

Acid hydrolysis yielded p-aminobenzoic acid, glutamic 
acid, alanine, leucine and perhaps another uncharac- 
terized substance (not a pteridine) with an U.V. maxi- 
mum at 360 m/t. 

A total synthesis of gramicidin Jo has been achieved. ^""^ 
The biosynthesis of this substance has been investigated.^*^ 
The antibiotic was concentrated in the RNA-rich proto- 
plast precipitate. 

The fact that bacitracin A (especially old samples) 
stimulates growth of Phycomyces blakesleanus may be 
due to conversion of the thiazoline ring of bacitracin A 
to a thiazole ring (bacitracin F)."^ 

Papers have appeared on metabolism and actinomycin 
production by streptomycetes'*^ and on the citric acid 
cycle and actinomycin formation. ^^"^ 

The cytostatic activity of actinomycins is reversed by 
high concentrations of purines and pyrimidines."" The 
interpretation of this effect was that actinomycin may 
react with DNA to form dye-polymer complexes. 

Mitomycin C causes bacteria to break down their DNA 
rapidly, acid-soluble products being formed. ^■*°'' 

An actinomycin complex, aurantin, colorless crystals, 
m.p. 255-257°, [ali,'*' - 308°, has been isolated in Rus- 
sia.^*' The complex contains threonine, sarcosine, pro- 
Une, valine, N-methylvaline and isoleucine. It was 
separated into four biologically active components: A^ 
m.p. 205°, A. m.p. 225°, A3 m.p. 226° and A4 m.p. 152°. 
Methionine furnishes the methyl groups attached to 
the aromatic chromophore of the actinomycins as shown 
by labeling wdth C^*.^*^° 
1" Y. Noda, J. Chem. Soc. Japan 80 411 (1959). 
142 s. Otani, I. Murakami and S. Chin, Abstr. 118th Meeting, 
Japanese Antibiotics Association. 

1*3 Sibor Ebringer, Naturwissenschaften 47 210 (1960). 
"* Paul Prave, Arch. Mikrobiol. 32 278 (1959). 
^*Udem., ibid. 32 286 (1959). 

1*6 W. Kersten, H. Kersten and H. M. Rauen, Nature 187 60 
(1960). 

i**"'" E. Reich, A. J. Shatkin and E. L. Tatum, Biochim. et Biophys. 
Acta 45 608 (1960). 

1*' A. B. Cilaev, T. I. Orlova, B. C. Kuznetsova and I. B. Mironova, 
Antibiotiki 3 18 (1960). 

>*■» A. J. Birch, D. W. Cameron, P. W. Holloway and R. W. Rick- 
ards. Tetrahedron Letters No. 25 26 (1960). 



Pfizer Handbook of Microbial Metabolites 694 

A general review of actinomycin structure and syn- 
thesis has appeared.^*'" 

A colorless, amorphous polypeptide antibiotic, edein, 
has been isolated from a strain of Bacilhis hrevis}*^ It 
contained arginine, glycine, glutamic acid, aspartic acid, 
tyrosine and two unidentified ninhydrin-positive spots. 

Two heat stable polypeptides, phytoactin and phy- 
tostreptin, have been isolated from an unclassified strep- 
tomycete.^*'' Both contain valine, a-alanine, proline, 
leucine or isoleucine, arginine, glycine and serine. 

Two peptide antibiotics not mentioned before are 
coliformin^'^" and roseocitrins A and B.^^^ Coliformin has 
the analysis: C 47.6, H 8.22, CI 3.31, S 0.23, P 0.47 and 
O 33.15 and contains alanine, glycine, serine, glutamic 
acid, aspartic acid, lysine, valine and leucine. The 
roseocitrins appear to resemble streptothricin. 

In a review of this class the name depsipeptide has 
been suggested for substances such as amidomycin and 
valinomycin, which are composed of a-hydroxy acids and 
amino acids. ^'^- Synthetic methods have been devised 
for both regular and irregular sequences of the two types 
of acids in these antibiotics. 

The biosynthesis of a,y-diaminobutyric acid in Bacillus 
circulans has been studied. ^^^ 

The structure of amidinomycin, C9HigON4 -112804, has 
been shown to be:^^^* 



O NH 

H2N- 

-C— NH— CHo— CHo— C— NH2-H2S04 



i*"''Hans Brockmann, Angew. Chem. 72 939-948 (1960). 

^''^ Z. Kurylo-Borowska, Symposium on Antibiotics, Prague, 1959. 

149 Jack ZifFer, S. J. Ishihara, T. J. Cairney and A. W. Chow, 
Phijtopathology 47 539 (1957). 

1^" Stig K. L. Freyschuss, Stig O. Pehrson and Borje Steinberg, 
Antibiotics and Chemotherapy 5 218 (1955). 

I'l Hisaya Kato, J. Antibiotics (Japan) 6A 143 (1953). 

152 M. M. Shemyakin, Angeiu. Chem. 72 342-345 (1960). 

1''^ Yelahanka Krishnamurthy Murthy, Dissertation, Purdue Univ., 
1958. 

'^^'■^^ Shoshiro Nakamura, Keiko Karasawa, Nobuo Tanaka, Hiroshi 
Yonehara and Hamao Umezawa, /. Antibiotics (Japan) 13A 362 
(1960). 



695 Addendum 

The structure of the antifungal antibiotic, variotin, 
CifiHo.O.N, is :'•"«" 

O 

II 
CHaCHj— CH.— CH C— CH =C— CH=CH— CH=CH— C— NH— CH,— CH,— CH> 

OH CHs 

O 

— C— OCH3 
Thus, while it is a tetraene, it is not of the macrolide 
class. 

Siderochromes. 

A number of microorganisms have been found to 
produce iron-containing pigments which absorb in the 
ultraviolet at 420-440 m^u, and have other properties in 
common. It has been suggested that these be called 
siderochromes.^^* 

Some of these substances are antibiotic and are called 
sideromycins. Others are growth factors and may be 
designated sideramines. The antibiotic sideromycins 
seem to function by inhibiting the growth factor sidera- 
mines. 

It remains to be seen how broadly the significance of 
these substances will extend. Some 50 strains of strepto- 
mycetes produce sideromycin-like antibiotics.^-* Of 32 
common microbial species examined 10 produced 
coprogen-like substances. ^^^ The sideramines seem to 
perform a coenzyme-like function in many microorgan- 
isms. 

Grisein A and albomycin have broad antibiotic activ- 
ity. In gram-positive microorganisms, but not in gram- 
negative ones, their effects are inhibited by sideramines. 
Ferrimycin is 10 to 50 times as effective as penicillin 
against gram-positive microorganisms in animal studies. 
The following table shows some of the siderochromes 
which have been best characterized: 
^^^" Setsuo Takeuchi, Hiroshi Yonehara, Hamao Umezawa and 
Yusuke Sumiki, ibid. 13A 289 (1960). 

^■* H. Bickel, E. Gaumann, W. Keller-Schierlein, V. Prelog, E. 
Vischer, A. Wettstein and H. Zahner, Experientia 16 129-133 
(1960). 

1"'= C. W. Hesseltine, A. R. WhitehlU, C. Pidacks, M. Ten Hagen, N. 
Bohonos, B. L. Hutchings and J. H. Williams, Mycologia 45 7 (1953). 



Of 




•o K 00 

lO lO "O 


o^ o 

IT) >o 


1 

— -o 
•o -o 


I 0- 


Ammonia, Succinic Acid 
1 -Amino-5-hydroxyl- 
aminopentane, 5- 
aminovaleric Acid, 
Cadaverine, Cryst. 
compound (X max. 
227, 323 m/u). Proline 
and 1 unidentified 
ninhydrin-positive 
substance. 


o i: 

o 

E X 

3 

^< 

5 


Methyluracil, Serine, 
Ornithine, Hydroxyl- 
amine 


Absorption 
X max. 

cm. 


228, 282 
319, 28 
425, 22 


00 
o &■ 

— CN 

lO o" 

-O CN 

CM -"l- 




a. 


00 00 
— 00 






1^ 


■o 
o 


CO 
o 


1 
o o 

CN CO 


s 

X 

o 

c 

< 


0) 
u. 






M3 


U 


o 
•d 






z 


o. 

CN 


CN 




I 


o- 

o 


<o 




u 


•o 

00 


O; 
CO 




Producing 
microorganism 


Sfreptomyces griseo- 
flavus (Krainsky) 
Waksman et Henrici, 
S. go/i/oeus, S. 
lavendulae 


S 5 
S X 

8 § 


Actinomyces subtropicus 
Kudrina et Kochet- 
kova 


'u 

>. 
E 
o 

4) 




Ferrimycin A (may 
consist of 2 com- 
ponents) 


< 

c 

'5 
O 


Albomycin (A com- 
plex. The main 
component has 
been resolved 
into two ports.) 



en. 




>o 




00 
>o 


o» 

"O 


o 


2 -D 

-0 o 
- a 


4t -? 

O X CO u 
c .S c '^ 

£ i J! 

1 c O < 
< 


X 

.1 ^ ^ § - 
JC 4, J, ii (L 

1 1 m 

° - -- 1: £2 

•c 2 o 1 ^ 
o 


< *« •- « 

:i t ^- "I < 

"^^ o S S 5; 

llllh 

1 - O < U < 

< 






Absorption 
X max. 

E.1% 

cm. 


o- 
CO 

>W 

CM 


CO 

o" 


CO 

<s 

CO 


•o 
CO 

o" 




a 












ii 


CN 


o 
o 


o 

N. 






» 



c 

< 


ID 




CO 




<> 
<) 











CN 






Z 


•o 


00 


CN 


•o 

CN 

d 




X 


o 

O; 


o 

00 


^ 

nI 


CO 

00 




u 


CM 
O 


IT) 

K 

■<1- 


o 
00 


■o 

O; 

d 
■o 




E 

1| 

= ? 

o 


I -E 

<U 0) 
D> > 
O <^ 
0) "S 
D ». 
-C 1= 

8- m 
X 

o 1 

^ = 
"^ CO 

3 


8 -E 

0) > 
o ^ 

0) % 
D VI 

S- Uj 
X 

i! 

^ = 
3 


O w «> 0) 3 5 

S 2 g • ui "E ? 
°' a i 3 £ ^ i 

s .. 1 § -s ;-. -5 S 

g 5 :i o „- "o -C 
a .i: . 3 g . . 


d. 

E 

c 

a. 


i 

0) 

J) 
£. 



■a 
o 


4) 

C 

1 
D 
0) 




o 

« 
11. 


< 

E 


u. 


CO 

« 

c 

"i 

o 

X 


« 

u. 


0) 

o 

Q. 

U 


o 

c 

O) 
4) 

« 

1- 



Pfizer Handbook of Microbial Metabolites 698 

Other less well characterized siderochromes were dis- 
cussed in reference 156. 

In the ferrichromes the iron is bound by coordination 
with hydroxamic acid derivatives of the ahphatic acid 
moieties. 



"6H. Bickel, E. Gaumann, W. Keller-Schierlein, V. Prelog, E. 
Vischer, A. Wettstein and H. Zahner, Experientia 16 128 (1960). 

15' H. Bickel, B. Fechtlg, G. E. Hall, W. Keller-Schierlein, V. Prelog 
and E. Fischer, Helv. Chim. Acta 43 901 (1960). 

^""^ H. Bickel, et al., to be published. 

159 D. M. Reynolds, A. Schatz and S. A. Waksman, Proc. Soc. Exp. 
Biol. Med. (New York), 64 50 (1947); D. M. Reynolds and S. A. 
Waksman, J. Bacterial. 55 739 (1948). 

160 F. A. Kuehl, M. N. Bishop, L. Chaiet and K. Folkers, /. Am. 
Chem. Soc. 73 1770 (1951). 

1*^1 M. G. Brazhnikova, N. N. Lomakina and L. I. Murayeva, 
Doklady Akad. Nauk. S.S.S.R. 99 827 (1954). 

162 E. O. Stapley and R. E. Ormond, Science 125 587 (1957). 

163 G. F. Gause, Brit. Med. J. 2 1177 (1955); G. F. Cause and 
M. G. Brazhnikova, Novosti Med. (Moscow) 23 3 (1951). 

164 Yu. O. Sazykin, Mikrobiologiya 24 75 (1955). 

165 E. S. Kudrina and G. V. Kochetkova, Antibiotiki (Moscow) 3 
63 (1958). 

166 O. Mikes and F. Sorm, Symposium on Antibiotics, Prague, 1959. 
16' J. B. Neilands, }. Am. Chem. Soc. 74 4846 (1952); idem., J. 

Biol. Chem. 205 643, 647 (1953); idem., Bacterial. Revs. 21 101 
(1957); J. A. Garibaldi and J. B. Neilands, /. Am. Chem. Soc. 77 
2429 (1955); Thomas Emery and J. B. Neilands, to be published; 
T. Emery and J. B. Neilands, Nature 184 1632 (1959). 

16S G. E. Hall, unpubhshed. 

169 C. W. Hesseltine, C. Pidacks, A. R. Whitehall, N. Bohonos, 
B. L. Hutchings "and J. H. Wilhams, /. Am. Chem. Soc. 74 1362 
(1952); C. W. Hesseltine, A. R. Whitehall, C. Pidacks, M. T. Hagen, 
N. Bohonos, B. L. Hutchings and J. H. Williams, Mycologia 45 7 
(1953); C. Pidacks, A. R. Whitehall, L. Pruess, C. W. Hesseltine, 
B. L. Hutchings, N. Bohonos and J. H. WiUiams, /. Am. Chem. Soc. 
75 6064 (1953). 

I'O A. G. Lochhead, M. O. Burton and R. H. Thexton, Nature 170 
282 (1952); A. G. Lochhead and M. O. Burton, Can. J. Botany 31 7 
(1953); M. O. Burton, F. J. Sowden and A. G. Lochhead, Can. J. 
Biachem. and Physiol. 32 400 (1954). 



699 



Addendum 



Baccatine A (entry 1114) has been shown to be a 
mixture of enniatins A and B.'"' 

A partial structure has been advanced for PA-114-B-1 
(entry 729).''- It is C^sHtjiOioNi,: 

OH 

/ 
C — L-Thre 

II I 

O O 

I 
0==C — L-a-Phenylglycine- 




"-N 



L-Proline 
Sarcosine 

p-Dimethylamino- 
phenylalanine 

5-Hydroxymethyl- 
-D-a-Aminobutyric Acid — hydroxyproline 
(perhaps) 

PA-114-B-3, a minor component of this synergistic com- 
plex, contains all the same components except sarcosine. 
It seems to contain another N-methyl amino acid instead. 
Other synergistic complexes of this sort are streptogramin, 
staphylomycin, ostreogrycin and mikamycin. These were 
classified as follows: 



Type 


Specific 
compound 


Synonyms 


A 


A, 


PA-n4-A-1 

Ostreogrycin (E-129) Factor A 

Mikamycin A 

Streptogramin main component 

Staphylomycin Mi 




A., 


Staphylomycin M^ 


B 


Bi 


PA-114-B-1 

Ostreogrycin (E-129) Factor B 

Mikomycin B 




Bo 


Staphylomycin S 




Bs 


PA- 114-8-3 






Streptogramin, minor component 



I'lG. E. HaU, Chem. and Ind., 1270 (1960). 

I'-D. C. Hobbs and W. D. Celmer, Nature 187 598 (1960). 



Pfizer Handbook of Microbial Metabolites 



700 



More data have been published on the purification and 
physical properties of mycobacillin (entry 795).^'^ 

Some degradation studies of thiostreptone (entry 809) 
have been reported/'* L-Threonine, L-isoleucine, l- 
alanine and D-cysteine were identified, and several thia- 
zole-containing fragments were isolated. A minimal 
molecular weight of 1500 is required. 

A structure has been proposed for a new antibiotic, 
racemomycin O.^"'^ It is produced by Streptomyces race- 
mochromo genus, has the empirical formula C25H44O10N8 
and is thought to be : 



HO CH. 

l/\ 
CH2 CH— C 

I I I 

HN N C 

\/ X\ / 
CO N 

I H 

NH 



NH 



\ 



HC 

HC— NH CO— CH2— CH— CH2— CH2— CH2— NH2 

I 
HOCH O NH2 

I 
HC— O 

I 
HC CH— CH2— CH2— O— CH2— CH— CH3 

CH2— O 1 OH 



A partial structure has been advanced for roseothricin 
A (entry 717).^"'' 



^" S. K. Majumdar and S. K. Bose, Arch. Biochem. and Biophys. 
90 154 (1960). 

^'* Miklos Bodanszky, John Timothy Sheehan, Josef Fried, Nina J. 
WiUiams and Carolyn A. Birkheimer, 7. Am. Chem. Soc. 82 4747 
(1960). 

i"S. Takemura, Chem. & Pharm. Bull. (Tokyo) 8 578 (1960). 

1^8 T. Goto, Y. Hirata, S. Hosoya and N. Komatsu, Bull. Chem. 
Soc. Japan 30 729 (1957). 



yoi 



Addendum 



— 


O— 

1 






r 


1 
CH, CH -C— CHo— NH— 

1 








NH N 

1 


1 
CO— 






2— CO— 


1 
NH 








— 0— 


HC 






-: 


— NH— 


HC 

1 


-NH- CO CH,^CH— CH,- 


-CH,— CH2— NH2 




— OH— 


— O— CH C 

1 


) NH.. 








— O— CH 










HC 

1 










1 
/-ij„ 








2 




^n2 






. 



A new polypeptide antibiotic, glumamycin, has been 
reported.^" It consists of colorless powder, m.p. 230° 
(dec), mol. wt. ^1800 and is composed of 4-isotri- 
decenoic acid, CsHi7CH=CHCoH4— COOH, L-aspartic 
acid, glycine, L-valine, L-proline, D-pipecolic acid and 
a,/3-diaminobutyric acid. 

A number of compounds listed in the unclassified sec- 
tion are known to be or thought to be polypeptides. 
These include alboverticillin, antibiotic B-456, bacilipins, 
bacillomycins, bacilysin, datemycin, diplococcin, dista- 
mycin A, laterosporin, melanosporin, mikamycins, mito- 
mycins, monilin, mycospocidin, phleomycin, pluramycins, 
racemomycins, ractinomycins, roseomycin, taitomycin, 
violacetin and undoubtedly others. 

18. Heterocycles 

C. PYRANS AND RELATED SUBSTANCES 

8-Hydroxy-3,4-dimethylisocoumarin has been isolated 
from cultures of a wild Oospora specimen.^"' 
^'' Michitaka Inoue, Hiroshi Hitomi, Komei Mizuno, Masahiko 
Fujino, Akira Miyake, Koiti Nakazawa, Motoo Shibata and Toshihiko 
Kanzaki, ibid. 33 1014 (1960). 

^'^ I. Yamamoto and Y. Yamamoto, Bull. Agr. Chem. Soc. (Japan) 
24 628 (1960). 



Pfizer Handbook of Microbial Metabolites 702 



A survey has shown that a-tocopherol is the only form 
of vitamin E found in bacteria."-' It was found in about 
a dozen chlorophyll-containing organisms, although not 
in all such bacteria which were studied. Its production 
is not limited to any particular type of chlorophyll-con- 
taining bacterium. Tocopherol production seemed to 
parallel chlorophyll production, and it was suggested that 
the same phytol precursor might be used for both. 

d. XANTHONES 

A labeled precursor investigation of ravenelin by Birch 
and associates has shown that orsellinic acid is an inter- 
mediate in the biosynthesis of xanthones.^®° 

e. COMPOUNDS RELATED TO THIOPHENE, IMIDAZOLE, 
THIAZOLE AND ISOXAZOLE 

A comparison of the effects of D-cycloserine and of d- 
alanine on the incorporation of D,L-alanine-l-C^^ into bac- 
terial proteins showed that D-cycloserine acts as a d- 
alanine antagonist. ^^^' ^'*- 

A paper has been published on the lability of 2-acetyl- 
thiazolium salts and in support of the proposed mode of 
action of thiamine. ^-^ 

A paper on the enzymatic formation of thiamine and 
phosphate esters of the pyrimidine moiety seems to be 
the first of a series on the biosynthesis of thiamine."* A 
dissertation on the biosynthesis of the thiazole moiety has 
been published."^ 

i"9 J. Green, S. Price and L. Gare, Nature 184 1339 (1959). 

180 Private communication from Herchel Smith. 

^^^ P. Barbieri, A. diMarco, L. Fuoco and A. Rusconi, Biochem. 
Pharmacol. 3 101 (1960). 

182 p Barbieri, A. diMarco, L. Fuoco, P. Julita, A. Migliacci and A. 
Rusconi, ibid. 3 264 (1960). 

1"-* Ronald Breslow and Edward McNeils, J. Am. Chem. Soc. 82 2394 
(1960). 

^>** Gerald W. Camiener and Gene M. Brown, J. Biol. Chem. 235 
2411 (1960). 

"^ J. Vogel, Dissertation, University of Bonn, 1960. 



703 Adden(lum 

f. PYRROLES, PORPHYRINS AND RELATED COMPOUNDS 

A dissertation has been published on a prodigiosin-like 
pigment.^'"' 

A partial synthesis of vitamin B,;. has been re- 
ported/^"' ^'^'^ 

Guanosine diphosphate factor B and B diphosphate ester 
have been indentified as intermediates in the biosynthesis 
of vitamin Bio.^"*" 

A dissertation has been published on the biosynthesis 
of members of the vitamin Bj^. group/"" '"^ 

A report has been made on the preparation and proper- 
ties of purified intrinsic factor. The purified material is 
a better Bjo binder than the crude, and it is not a muco- 
protein as previously believed.^®* 

A publication on the biosynthesis of uroporphyrin III 
from porphobilinogen reported that uroporphyrinogen I 
is not an intermediate in the biosynthesis of uroporphy- 
rinogen UW 

A pink pigment identified as coproporphyrin III was 
isolated from Mycobacterium tuberculosis avium^^* as it 
had been earlier from Mycobacterium karlinski}^^ 

At least two kinds of chlorophylls have been shown to 
be present in green bacteria.'"*^ 

^^•^ Roswltha Zimmer-Galler, Dissertation, Technische Hochschule, 
Miinchen, 1960. 

^'*' K. Bernhauer, F. Wagner, Hw. Dellweg and P. Zeller, Helv. 
Chim. Acta 43 700 (1960). 

iss-yy Friedrich, G. Gross, K. Bernhauer and P. Zeller, ibid. 43 

704 (1960). 

i**^ G. Boretti, A. dlMarco, L. Fuoco, M. Marnatl, A. Migliacci and 
C. Spalla, Biochim. et Biophys. Acta 37 379 (1960). 

1"" Fred Sanders, Dissertation Abstr. 18 2189 (1959). 

1"^ F. Sanders and Gerald R. Seaman, /. Bacteriol. 79 619 (1960). 

"- Leon Ellenbogen and William L. Williams, Biochem. and 
Biophys. Res. Comms. 2 340 (1960). 

^''■' Lawrence Bogorad and Gerald S. Marks, Biochim. et Biophys. 
Acta 41 358 (1960). 

^^D. S. P. Patterson, Biochem. J. 76 189 (1960). 

195 c. M. Todd, ibid. 45 386 (1949). 

^^^ R. Y. Stanier and J. H. C. Smith, Biochim. et Biophys. Acta 41 
478 (1960). 



Pfizer Handbook of Microbial Metabolites 704 

A b-type cytochrome has been isolated from the fungus 
Sclerotiana libertiana and identified. ^^^ 

Protoporphyrin IX has been isolated from bacterial 
catalase and characterized. ^^^ 

Addition of S-aminolevuHnic acid to cultures of pro- 
pionibacteria caused large increases in the production of 
porphyrins, but no rise in vitamin Bjo production, in- 
dicating divergent biosynthetic routes."^ 

The structure of the antifungal pigment prodigiosin 
has been proved by synthesis. -°° It is 



OCH 




and is thus the second natural product containing a 2,2'- 
dipyrrole skeleton, vitamin Bjo being the other. 

A streptomycete has been found w^hich produces iso- 
butyropyrrothine, orange-red antibiotic crystals, m.p. 



228 



o . 200a 



CH3 

/ 

S- -C=C— NH— CO— CH 

sec CH3 

C N O 



H 



CH3 



Aureothricin, thiolutin, a colorless base, and a heptaene, 
hamycin, also were produced. 

^^" Tateo Yamanaka, Takehazu Horio and Kazuo Okunuki, Biochim. 
et Biophys. Acta 40 349 (1960). 

1"* Steve Miller, Davis Hawkins and Robert P. Williams, /. Biol. 
Chem. 235 3280 (1960). 

1^^ G. V. Pronyakova, Biokhimiya (English translation) 25 223 
(1960). 

'°° Henry Rapoport and Kenneth G. Holden, /. Am. Chem. Soc. 82 
5510 (1960). 

2oo« D. S. Bhata, R. K. Hulyakar and S. K. Menon, Experientia 16 
504 (1960). 



705 Addendum 

g. INDOLES 

The structure previously proposed for echinulin has 
been confirmed, the only reservation being possible ex- 
change of the groups in the 5 and the 7 positions of the 
indole nucleus.-''^ 

926a Lysergic Acid Amide (Ergine), CieHiyONa (Monomethanolate), 
m.p. 130-135° (efferv.), resolidifies 140°, m.p. 190° with 
previous dec. 

926b Isolysergic Acid Amide (Isoergine), CicHi-ONg. 

937a Lysergic Acid Methylcarbinolamide, CisHo^OoNg, colorless 
prisms, m.p. 135° (dec), [aW +29° ±2° (--1.0 in 
dimethylf ormamide ) . 

937b Isolysergic Acid Methylcarbinolamide, C18H21O0N3, not crystal- 
Une. 

A yield of about 2 g. per liter of the above alkaloids was 
produced by Claviceps paspali Stevens T. Hall growing in 
submerged culture.-"- A partial structure is shown for 
the carbinolamide isomer corresponding to lysergic acid: 

CH3 

NH— C— OH 

/ I 
0=C H 



O 



N— CH3 



Another new ergot alkaloid, molliclavine, has been 
reported : -"^ -"* 

-°^ Franco Piozzi, Giuseppe Casnati, Adolfo Quilico and Cesare 
Cardani, Gazz. chim. ital. 90 451, 476 (1960). 

2"- F. Arcamone, C. Bonino, E. B. Chain, A. Ferrettl, P. Pennella, 
A. Tonola and Lidia Vero, Nature 187 238 (1960). 

^°^ M. Abe, S. Yamatodani, T. Yamano and M. Kusumoto, /. Agr. 
Chem. Soc. Japan 34 249 (1960). 

^''^M. Abe and S. Yamatodani, Bull. Agr. Chem. Soc. (Japan) 19 
161 (1955). 



Pfizer Handbook of Microbial Metabolites 706 

930a Molliclavine, CieHjgOoNo, colorless crystals, m.p. 253° (dec), 
[a]i>'' +30° (c 1.0 in pyridine). 




Claviceps purpurea 

An antibiotic of novel structure incorporating an indole 
nucleus is: 

391b PA-155A, C14H15O2N3, colorless crystals, m.p. 209°, [ajp^^ -214° 
(c 2.0 in methanol), U.V. 218, 273, 281, 288 m/x. 

No reaction with dinitrophenylhydrazine. Negative 
ninhydrin, FeCl^ tests. Blue Ehrlich's test. Decolorizes 
Br2, KMn04. Streptomyces alhus''^' "'^^ 

i. PYRIDINES 

The plant toxin, fusaric acid, was produced when 
Fusarium oxysporum var. lini was grown on artificial 
medium or on non-resistant flax tissues, but not when 
the fungus was grown on resistant strain tissues. -°'^ 

A dissertation has been pubUshed on dipicolinic acid 
formation and other chemical aspects of bacterial sporula- 
tion.-"^ 

The mononucleotide of nicotinic acid has been isolated 
from a fusarium specimen^°^ and from a yeast.-"* 

k. PYRAZINES, DIKETOPIPERAZINES 

Several diketopiperazines have been isolated from the 
fungus Rosellinia necatrix Berlese.-"'' They are L-prolyl- 

^•'' Koppaka V. Rao, Antibiotics and Chemotherapy 10 312 (1960). 
''^" Manfred von Schach, private communication. 
205 E. J. Trione, Phytopathologij 50 480 (1960). 
2''« Herbert M. Nakata, Dissertation Abstr. 20 3020 (1960). 
2"' A. Ballio and S. Russi, Arch. Biochem. and Biophys. 85 567 
(1959). 

2'« R. W. Wheat, ibid. 85 567 (1957). 

2"»Yu-Shih Chen, Bull. Agr. Chem. Sac. (Japan) 24 372 (1960). 



70? Addendum 

L-leucine anhydride, L-prolyl-L-valine anhydride and an 
apparently new diketopiperazine, L-prolyl-L-phenylalanine 
anhydride (compound E) Ci^Hi.jO.jN.j, m.p. 127-128°, 
[a]..-'" —99.8" (c 1.0 in ethanol). A crystalline wax, m.p. 
52°, was isolated from the same culture and assumed to 
be n-pentacosane, Co-H-.o. Also an uncharacterized sub- 
stance, white needles, m.p. 206-208°, called rosellinic 
acid was isolated. 

L-Prolyl-L-valine anhydride had been isolated previously 
from a streptomycete culture.-^" L-Prolyl-L-leucine anhy- 
dride had been isolated both from a streptomycete and 
from Aspergillus funiigatus.'^^ 

Muta-aspergillic acid, C11H18O3N2, pale yellow needles, 
m.p. 173° (dec.) (subl.) with alternative structures: 



CH3 




CH3 


/ 




i 


CHo— CH 




C— CH; 


ch| j CH3 


or CH3 

\ 


1 J O" 


CH- 


-CHo 1 ^0 


1 OH 


/ 


OH 


OH 


CH3 





has been reported.-"' 

/. PHENAZINES AND PHENOXAZONES 

Three new natural phenazines have been reported. ^^^ 

984b l-Hydroxymethyl-6-carboxyphenazine, C15H12O3N2, light yellow 
crystals, m.p. 197-201°. 

HOOC 

I 



.V 



CH2OH 

210 Y. Koaze, ibid. 22 98 (1958). 

-" J. L. Johnson, W. G. Jackson and T. E. Elbe, ] . Avn. Chem. Soc. 
73 2947 (1951). 

2"' Seiji Nakamura, Bull. Agr. Chem. Soc. (Japan) 24 629 (1960). 
2i2Koki Yaglshita, J. Antibiotics (Japan) 13A 83 (1960). 



Pfizer Handbook of Microbial Metabolites 708 

985a l-Methoxy-4-methyl-9-carboxyphenazine, C16H14O3N2, yellow 
needles, m.p. 124-126°. 



HOOC 




984a l-Methoxy-4-hydroxyniethyl-9-carboxyphenazine (Griseolutic 

Acid) C15H12O3N. 




CH2OH 



All of these compounds were Isolated from a culture of 
Streptomyces griseoluteus. 

An unclassified streptomycete produced two substances 
which were named questiomycins A and B.-^^ These 
have been identified as : 

977a 6-Aminophenoxazone (Questiomycin A) CisHgOgNo, red crys- 
tals, m.p. 241-244° (dec.) subl. from 150°. 

NH2 



o 

377a 2-Aminophenol (Questiomycin B), colorless crystals, m.p. 170- 
175° (subl. 120°). 

NH2 



OH 

The suggestion was made that the aminophenol might be 
the precursor of the aminophenoxazone. 

A purple and a yellow pigment isolated from Brevibac- 
^^^ Kentaro Anzai, Kiyoshi Isono, Kazuhiko Okuma and Saburo 
Suzuki, ibid. 13A 125 (1960). 



yog Addendum 

teniim crystalloiodimim Sasaki, Yoshida et Sasaki have 
been identified as iodinin and 1,6-dihydroxyphenazine, 
respectively.-^* 

m. PYRIMIDINES 

Two dissertations have been pubhshed on the bio- 
synthesis of pyrimidines, one with rat Uver enzymes,^^' 
the other with Neurospora crassa.'^^'^ 

Thymidine diphosphate mannose (as well as the pre- 
viously reported thymidine diphosphate rhamnose) has 
been isolated from cultures of Streptomyces griseus.^^'' 

It is possible that this substance is an intermediate in 
the biosynthesis of streptomycin B (o(-T>-mannopyranosyl- 
streptomycin) which is produced by this organism along 
with streptomycin. 

Tritium labeling experiments indicated that in the 
case cited, at least, the epimerization of N-acetylglu- 
cosamine to N-acetylmannosamine, probably by way of 
uridine diphosphate N-acetylglucosamine, does not in- 
volve oxidation to a ketosugar, followed by stereospecific 
reduction. -^^^ 

The structure of tubercidin, C11H14O4N4, m.p. 247'' 
(dec), [alo^' —62°, produced by Streptomyces tubercidi- 
cus and active against Mycobacterium tuberculosis and 
Candida albicans, has been reported to be:-^**" 

NH2 

I 

4-amino-7-[D-ribofuranosyl]- 
pyrrolo-[2,3-d]-pyrimidine 



^ 



-N-^ 



D-ribose 
Toyocamycin has a similar structure. ^^^^ 

2^*Tosi Irle, Etsuro Kurosawa and Iwao Nagaoka, Bull. Chem. 
Soc. Japan 33 1057 (1960). 

215 Richard L. Stambaugh, Dissertation Abstr. 20 64 (1959). 

2i6Kamala P. Chakraborty, ibid. 20 3044 (1960). 

21" J. Baddiley and N. L. Blumson, Biochim et Biophys. Acta 39 
376 (1960). 

218 Luis Glaser, ibid. 41 534 (1960). 

218" Saburo Suzuki and Shingo Marumo, J. Antibiotics (Japan) 
13A 360 (1960). 

218" Kazuhiko Ohkuma, ibid. 13A 361 (1960). 



Pfizer Handbook of Microbial Metabolites 710 

72. PURINES 

Guanosine diphosphate glucose and guanosine diphos- 
phate fructose are produced by Eremotheciiim ashbyii.^^^ 
A dissertation reports the isolation of a new guanine de- 
rivative from a riboflavin producer.""" 

A new purine riboside has been isolated from fusarium 
species.'-^ It has been assigned the provisional struc- 
ture: 

2-( 1 -Carboxyethylamino )-6-hydroxy-9-D-ribofuranosyl- 
purine 

OH 






CH;,— CH— HN 
COOH 
H:03P— O— CH, 




OH OH 

Nebularine (9-/?-D-ribofuranosylpurine), produced by 
Agaricus nebularis, has been isolated from a strepto- 
mycete.--^" 

The nucleotides of Aspergillus oryzae have been char- 
acterized. --- 

The mechanism of action of the antibiotic, psicofura- 
nine, against Staphylococcus aureus has been studied. ^'^ 
A possible effect may be interference with the biosynthe- 
sis of guanylic acid from xanthylic acid. 

219 H. G. Pontis, A. L. James and J. Baddiley, Biochem. J. 75 428 
(1960). 

220Usama A. S. Al-Khahdi, Dissertation Abstr. 21 (1960). 

"1 Alessandro Ballio, Carlo Delfini and Serena Russi, Nature 186 
968 (1960). 

'"'" Kiyoshi Osono and Saburo Suzuki, /. Antibiotics (Japan) ISA 
270 (1960). 

*'-- Kazuo Okunuki, Kozo Iwasa, Fumio Imamoto and Tadayoshi 
Higashiyama, /. Biochem. (Tokyo) 45 795 (1958). 

"•''Ladislav J. Hanka, J. Bacterial. 80 30 (1960). 



yii Addendum 

The antibiotic, mitomycin C, blocks DNA synthesis 
completely in EschericJiia coli, but does not interfere with 
RNA synthesis or protein synthesis."' Phage-infected 
bacteria continued DNA synthesis, but no infective par- 
ticles were produced when high concentrations of mito- 
mycin were present. 

A new incompletely characterized electron transport 
component has been isolated from Mjicohacterium phleip-^ 

The mode of inhibition of electron transport by anti- 
mycin A has been studied.--'' 

Evidence has been published for participation of a vic- 
dithiol in oxidative phosphorylation.'--' 

A review of ion transport and respiration has been pub- 
lished."'' 

ATP can replace light in bacterial photosynthesis. This 
discovery was made with the use of the obligate photo- 
troph chromatium. An acetate medium is adequate, and 
carbon dioxide is not required. ^-^ 

The biosynthesis of nucleic acids has been reviewed. ^^^ 

The biosynthesis and interconversions of purines and 
their derivatives have been reviewed. ^^^ 



0. PTERIDINES AND FLAVINES 

The prosthetic group of a chromoprotein from myco- 
bacteria may be a pteridine.^"*^ 

In the fly, Drosophila melanogaster, labeling studies 
indicate that glucose carbon atoms are specifically in- 

--■* M. Sakiguchi and Y. Takagi, Biochim. et Biophys. Acta 41 434 
(1960). 

--=W. B. Sutton, Federation Proc. 19 31 (1960). 

"«A. L. Tappel, Biochem. Pharmacol 3 289 (1960). 

"' Arvan Fluharty and D. R. Sanadi, Proc. Nat. Acad. Sci. U.S.A. 
46 608 (1960). 

228 R. N. Robertson, Biol. Revs. 35 231-265 (1960). 

22^ M. Losada, A. V. Trebst, S. Ogata and Daniel I. Arnon, Nature 
186 753 (1960). 

2'"' Arthur Romberg, Reviews of Modern Physics 31 200-209 
(1959). 

-•''^ Albert G. Moat and Herman Friedman, Bacteriol. Revs. 24 309 
(1960). 

232 F. B. Cousins, Biochim. et Biophys. Acta 40 532 (1960). 



Pfizer Handbook of Microbial Metabolites 712 

corporated into pteridines, but not into purines produced 
by the organism. ^^^ 

The structure of "active formaldehyde" (N^N^°-methyl- 
enetetrahydrofolic acid) has been proved by synthesis. ^^* 

19. Unclassified Metabolites 

Streptolydigin probably contains 4 carbon-carbon dou- 
ble bonds conjugated v\dth a ^-diketone system. -^'^ It also 
contains at least four hydroxyl groups, at least four C- 
methyl groups and at least one amide group. Methyla- 
mine was a base hydrolysis product of tetradecahydro- 
streptolydigin. 

Griseoviridin, empirical formula C22Ho9±207N3S, prob- 
ably consists of three moieties. ^^'^ A 6 carbon atom frag- 
ment has been identified as: 




and the sulfur atom may be attached at the X-position. 

The probable structure of a methanolysis product of 
carzinophilin has been published. ^^^ It is: 

Methyl 1 -methyl- 7-methoxynaphthalene-6-carboxylate 



CH3 



CH3O 



CH3OOC 

Mikamycin should be classified as a polypeptide of the 
etamycin type. L-Proline and glycine have been char- 
acterized in a hydrolysate. A monoacetate, a di-2,4- 
dinitrophenylhydrazone derivative, and a decahydro de- 

"3 O. Brenner-Holzbach and F. Leuthardt, Helv. Chim. Acta 42 
2254 (1959). 

-•^* M. J. Osborn, P. T. Talbot and F. M. Huennekens, J. Am. Chem. 
Soc. 82 4921 (1960). 

-^'^ Jerome Allen Course, Dissertation, Univ. of Illinois, 1959. 

236 p. de Mayo and A. Stoessl, Can. J. Chem. 38 950 (1960). 

237 Masao Tanaka, Teruo Kishi and Yoshiki Maruta, /. Antibiotics 
(Japan) 12B 361 (1959). 



713 Addendum 

rivative have been prepared. The melting point of the 
yellow crystals is given as 178° (dec.)-^" 

Russian antibiotic 6613 may be identical with eta- 
mycin.^^^ 

Monamycin, CooH3f5_3sO..^N, needles, m.p. 126°. Mono- 
hydrochloride: m.p. 187°, [a],/^ -62 ±5° (c 0.9 in 
ethanol), containing 1 N — CH;,, 3 C — CH;^ groups, no 
U.V., I.R. suggestive of amide links, has been isolated 
from Streptomyces jamaicensis n. sp.-*° 

Teruchiomycin, Co.sH^^OioN, needles, m.p. 202-204° 
(dec), a new antibiotic from Streptomyces antibioticus 
has been reported."*^ 

A new acidic antibiotic, C-159, U.V. max. 345, 260, 
280 mix, C 58.7, H 7.4, D 24.0, N 9.9% has been pat- 
ented.-^- 

The blue intracellular pigment of Pseudomonas lemon- 
nieri has been isolated, purified and characterized.^*^ 

A preliminary investigation has been made of the pig- 
ments of Trichophyton rubrum.^** 

Rubidin, a quinoid dark red powder with acid base 
indicator properties, U.V. 320, 415, 490 m^x in butanol, 
C 51.9, H 5.56, O 42.54, positive FeCly and zinc dust 
tests, is a substance isolated from an unclassified strep- 
tomycete.-*^ 

A new antibacterial antibiotic has been reported. ^*^ It 
had the following properties: yellow needles, m.p. 134°, 
mol. wt. 397, U.V. maxima at 328.5, 314.5, 298.7 m/x. 
Positive Millon, Liebermann, Schiff, FeClg and NH3- 
AgNO^ tests. Probably C22H22O7 with hydroxyl, methyl 
and 2 ketone groups present. 
238Koichi Okabe, ibid. 12A 86 (1959). 

239 M. Brazhnikova et al., Antibiotics (USSR) 4 414 (English 
translation) (1959). 

2*°C. H. Hassall and K. E. Magnus, Nature, Suppl. 184 1223 
(1959). 

241 H. Umezawa et al., Japanese Patent 850 (1958). 
2*2 British Patent 814,794 (1959). 

243 Werner Blau, Gladys Cosens and Mortimer P. Starr, Bacteriol. 
Proc, 153 (1960). 

2^Malati Bacchwal and G. C. Walker, Can. J. Microbiol. 6 383 
(1960). 

245 A. K. Banerjee, G. P. Sen and P. Nandi, "Antibiotics Annual 
1955-1956," Medical Encyclopedia, Inc., New York, p. 640. 

246Thadee Staron and Albert Faivre-Amiot, Compt. rend. 250 1580 
(1960). 



SUBJECT INDEX 

Bold-faced numbers indicate primary microbial metabolite 
entries, while Arabic numbers signify incidental mention under 
such entries. Italic numbers are page numbers, and generally 
indicate occurrence in a chapter or section introduction. The 
appendixes and addendum are not indexed. 



Abikoviromycin, 1183 

Aburamycin, 1064 

Acetaldehyde, 14, 17, 72, 466, 

480 
Acetate, 17, 48, 52, 80, 81, 91, 

120, 144, 154, 155, 159, 

160, 182, 187-189, 212, 

232, 236, 239, 273-275, 299, 

312, 398, 400, 420, 447, 555 
2-C^*- Acetate, 182,274 
Acetic acid, 17, 46, 69, 72, 82, 275 
l-C^^- Acetic acid, 159, 182 
Acetic acid (C"-labeled), 233, 

236, 411 
Acetoacetate, 80, 93, 190 
Acetoacetyl coenzyme A, 17, 93, 

155 
Acetoin, 15, 17, 19, 557-560 
a-Acetolactic acid, i5, 3i5 
Acetomycin, 82, 150 
Acetone, 18, 466 
Acetopyrro thine, 914 
4-Acetoxycycloheximide, 304, 309, 

316 
4-Acetoxyheximide, 305 
Acetyl coenzyme A, 15-17, 47, 

48, 53, 54, 93, 155, 424, 447 
Acetylcholine, 466, 654 
2-Acetyl-2-decarboxamidooxytet- 

racychne, 275, 612 
5-Acetyldihydrolipoic acid, 16 
0-Acetyleburicoic acid, 360 
Acetylenedicarboxylic acid, 108 
Acetylenic acids, JOS, J 09 

compounds, 107, 427 
N-Acetyl-D-glucosamine, 344, 345 
6-0-Acetylglucose, 37 
N-Acetylmuramic acid, 343 
N-Acetylneuraminic acid, 344 



N-Acetyltyramine, 407 

Achromycin, 613 

Acidomycin, 899 

Aconitase, 46 

cis-Aconitic acid, 47, 49, 92 

Actidione, 308 

Actilin, 63 

Actinobolin, 1065 

Actinochrysin, 764 

Actinocinin, 335, 336, 502 

Actinoleukin, 1066 

Actinomycin, 123, 742, 764, 770 

Actinomycin I, 805 

II, 811 

III, 812 

IV, 794 

V, 803 

VI, 795 

VII, 793 
Actinomycin B^, 794 

B.., 803 
C„ 794 

C, 795 
C.,„ 795 

C3, 336, 338, 793 

D, 794 
El, 796 
Eo, 797 
Fi, 798 
Fo, 799 
F,, 800 
F4, 801 
Ii, 794 
Jo, 12 

nomenclature, 381, 382 
Xo^, 805 

X„. , 806 
Xo,„ 807 
Xi, 794 



Pfizer Handbook of Microbial Metabolites 



716 



Actinomycin Bi 
Xia, 802 
Xo, 803 
X3, 804 
Zn, 808 
Zi, 809 
Actinomycins, 334-338, 381, 
382, 1001, 1132, 1214, 1250, 
1260 
Z, 808 

Zo, Z3, Z4, 809 
Z„ 810 
Actinorhodin, 234, 526, 529 
Actinorubin, 737 
Actiphenol, 306 
Actithiazic acid, 426, 899 
Active acetaldehvde, 315, 423, 
559, 560 
amino acids, 534 
carbon dioxide, 526 
formaldehyde, 549, 552, 553 
formate, 549, 551 
succinate 312,423 
Active sulfate, 524, 525 
Acyl adenylates, 525 
Acyl coenzyme A, 525 
Acyldehydrogenase, 53, 92 
Adenine, 318, 442, 445, 483, 
508-510, 529, 551, 559, 
1026, 1044 
Adenine-S-C^*, 557, 558 
nucleoside, 445, 535 
nucleotide, 526, 527 
Adenosine, 1033 

diphosphate (ADP), 14, 47, 
54, 55, 92, 333, 450, 524, 
530, 531, 535, 536, 560, 
562, 564, 1038 
diphosphoryl biotin, 55 
-2'-phosphate, 1036 
-3'-phosphate, 1037 
-5'-phosphate (AMP), 53, 318, 

510,530, 533, 1038 
-3'-phospho-5'-phosphosulfate, 

524, 525 
triphosphate (ATP), 14, 47, 
53-55, 92, 93, 291, 311- 
313, 318, 333, 345, 425, 
450, 511, 514, 515, 524- 



Adenosine 

526, 530, 53 J, 533, 535- 

537, 1040 
-5'-triphosphate, 1040 
5-Adenosylhomocysteine, 553 
S-Adenosylmethionine, 31 J, 525, 

553 
Adenosyl-5'-phosphoryl carbonate, 

526 
Adenylic acid, 345, 533 
Adenylic acid a, 1036 
3-Adenylic acid, 1037 
Adenylic acid-pantoate complex, 

334 
Adenylo-p-aminobenzoic acid, 556 
Adenylosuccinic acid, 533, 1044 
Aerosporin, 780 
Agaric acid, 120 
Agaricic acid, 49, 120 
Agaricin, 120 
AgaricoHc acid, 355 
Agmatine, 466 
Agroclavine, 471, 944 
Agrocybin, 190 
Akitamycin, 1067 
Aklavin, 616 
Alanine, 290, 300-302, 304, 305, 

309, 340-343, 435, 497, 501, 

725, 756, 757, 766, 769, 

773, 789, 813, 815-818, 822, 

828, 829, 831, 839-841, 849, 

1079 
/?- Alanine, 300, 303, 309, 310, 

333, 470, 535, 666, 726 
D-Alanine, 310, 343, 345 
L-Alanine, 343, 665, 704, 790 
D-Alanine-D-glutamate aminopher- 

ase, 488 
D-Alanyl-D-alanine, 345, 422 
Alazopeptin, 725 
Albamycin, 885 
Albidin, 1068 
Albofungin, 1069 
Alboleersin, 579 
Albomycetin, 1070 
Albomycin, 765, 766 
Alboverticillin (hydrochloride), 

1071 
Alcohol, 15, 17 



717 



Subject Index 



Alcohol dehydrogenase, 13 

Alcohol fermentation (yeast), 13 

Aldehydes. 564 

Aldolase, 13 

Aldol condensations, 16 

Alectoronic acid, 187 

Alicyclic compounds, 142 

Aliomycin, 1072 

Alkaloid biosynthesis, 459, 467- 

472 
Allantoic acid, 672 
Aliomycin, 1022, 1073 
Allophanic acid, 55 
Alternaric acid, 116 
Alternarine, 1074 
Alternariol, 151, 185, 414, 419, 
420 
methyl ether, 151, 415 
Altenuic acid I, 151,420 

II, 151, 421 

III, 151, 422 
Altenusin, 151, 419 
Altertenuol, 151, 416 
Althiomycin, 1075 
Alvein, 830 
a-Amanitin, 756 
/?-Amanitin, 756 
y-Amanitin, 756 
Amaromycin, 259 
Amebacillin, 318 
Amethopterin, 422 
Amicetin, 21, 346, 671, 1022 

B, 1020 
Amide, 922 

Amidomycin, 747-750, 758, 767 
Amines, 290, 458, 564 
Aminoacetone, 642 
Amino acid decarboxylase, 485 
Amino acid from Lactarius hel- 

vus, 710 
Amino acid racemase, 485 
Amino acids, 284, 290, 299, 508 
D-Amino acids, 345, 564 
Amino acids (activated), 345 
Amino acids (intracellular), 304, 

305 
Amino acid transport, 488 
a-Aminoadipic acid, 301, 312 
L-a-Aminoadipic acid, 694, 724 



D-a-Aminoadipic acid, 421, 911 
a-Aminoadipic acid e-semialde- 

hyde, 3J2 
S- ( a-Aminoadipyl ) cysteinylvaline, 

42 J, 724 
p-Aminobenzoic acid, i43, 53 J, 

556, 557, 699, 1059 
p-Aminobenzoylglutamic acid, 556 
a-Aminobutyric acid, 341, 739, 

751, 755 
y-Aminobutyric acid, 300, 303, 

342, 501, 673, 829 
D-a-Aminobutyric acid, 704, 755 
L-( + )-3;-Aminobutyric acid, 674 
l-Amino-3,6-desoxyhexose, 29J 
2-Amino-4,7-dihydroxypteridine- 

6-acetic acid, 1049 
3-Amino-l ,8-dimethylphenoxan-2- 

dicarboxylic acid-4,5, 788 
/:?-Aminoethanethiol, 535 
2-Aminohexose reactions, 23, 64 
2-Amino-4-hydroxy-6-hydroxy- 

methylpteridine, 556 
2-Amino-4-hydroxypteridine-6- 

carboxaldehyde, 556 
4-Amino-4-imidazolecarboxamide 

riboside, 53 J, 898 
5-Amino-4-imidazolecarboxamide 

ribotide, 551 
5-Amino-4-imidazolecarboxylic 

acid ribotide, 53 i 
Aminoimidazole ribotide, 53i 
5-Amino-4-imidazole-N-succino- 

carboxamide ribotide, 53i 
a-Aminoisobutyric acid, 726 
D-4-Amino-3-isoxazolidone, 894 
8-Aminolevulinate synthetase, 485 
5-Aminolevulinic acid, 435, 437, 

444, 550 
a-Aminomethyl-o!,/?-trans-,y,8-cis- 

muconic acid, 483 
2-Amino-4-methyl-5-oxy-3-pente- 

noic acid, 756 
2-Amino-4-methyl-3-pentenoic 

acid, 757 
l-Amino-2-methyl-2-propanol, 649 
2-( l-Amino-2-methylpropyl) 

thiazole-4-carboxylic 

acid, 762 



Pfizer Handbook of Microbial Metabolites 



718 



2- Amino-6-oxy purine, 508 
6-Aminopenicillanic acid, 418, 

419, 421, 897 
a-Amino-^-phenylbutyric acid, 760 
5- Amino- 1 -d- ( 5'-phosphoribosyl )- 

4-imidazolecarboxamide, 318 
6-Aminopurine, 508 
l-Aminoribose-5'-phosphate, 530 
p-Aminosalicylic acid, 53J 
Amino sugars, 22, 120, 308 
Ammonia, 290, 291, 308, 309, 

466, 515, 533, 637, 729-31, 

762 
Amosamine, 21 
Amphomycin, 833, 835 
Amphotericin, 249 
Amphotericin-A, 122, 233 
Amphotericin-B, 20, J 22, 248 
iso- Amylamine, 466 
Amytal, 449 

Anaerobic glycolysis, 13, 15 
Anasterol, 333 
Aneurin, 903 
Aneurindiphosphate, 904 
Angolamycin, 291 
Angustmycin A, 21, 1041 

C, 1042 
N\N^"-Anhydroformyl tetrahydro- 

folic acid, 530 
Aniline, 502 

Anisaldehyde, 284, 427, 619 
Anisic acid, 284 
Anisomycin, 1076 
Anthranilic acid, 143, 186, 317, 

458, 460, 492, 493, 502, 698 
Anthranols, 232 
Anthraquinone pigment from Gib- 

berella fujikuroi, 534 
Anthraquinones, i85, 190, 212, 

231-233, 254, 273 
bis-Anthraquinones, 2 J 4, 234 
Anthrones, 232 
Antibiotic 26/1, 1097 
Antibiotic 289, 577 
Antibiotic 446, 1098, 1197 
Antibiotic 587 13, 1100 
Antibiotic 720-A, 1099 
Antibiotic 899, 832 
Antibiotic 1037, 1101 



Antibiotic 1968, J 22 
Antibiotic 6270, 1102 
Antibiotic 6706, 1103 
Antibiotic A 246, 229, 1077 
Antibiotic B-456, 1078 
Antibiotic C-159, 1079 
Antibiotic D-13, 1080 
Antibiotic E-212, 1081 
Antibiotic E.F. 185, 63 
Antibiotic from Bacillus cepae, 

1090 
Antibiotic from Bacillus pumilis, 

1091 
Antibiotic from Monosporium bo- 

norden, 1092 
Antibiotic from Penicillium spin- 

ulosum, 1093 
Antibiotic from Streptomyces abi- 

koensis, 1094 
Antibiotic from Streptomyces fun- 

gicidicus, 1095 
Antibiotic from Streptomyces gris- 

eus, 1096 
Antibiotic HA-9, 1295 
Antibiotic I.C.I. 13,959, 726 
Antibiotic LA-7017, 1082 
Antibiotic M-4209, 1083 
Antibiotic PA-93, 885 
Antibiotic T, 1085 
Antibiotic X-206, 1086 
Antibiotic X-340, 611 
Antibiotic X-464, 1087 
Antibiotic X-465A, 439 
Antibiotic X-537A, 1088 
Antibiotic X-1008, 1089 
Antibiotic Y, 828 
Antibiotic Yo, 829 
Antibiotic from yeast, 828, 829 
Antifungal substance, 1104 
Antifungal substance produced by 

Streptomyces strain No. 1037, 

1105 
Antimycin A, 238, 449, 848 
A, 269 
A.,„, 270 
A„„ 271 
A,, 272 
A4, 273 
Antimycoin, 122, 237 



719 

Anziaic acid, 477 

Aquamycin, 5 

d-Ai-abi'tol. 22 

Arachidic acid, 50 

Arachidonic acid. 5 J 

Arginine, 300, 301, 303, 305, 308, 
309, 340-342, 821, 822, 824, 
830, 844, 845, 1145 

L-Arginine, ()96 

Argininosuccinate, 308 

Argomycin, 110() 

Aromatic amino acids, 143 
compounds, 286 

Ascorbic acid, 79, 82, 143, 460 
biosynthesis, 82 

Ascorbigen, 460 

Ascosin, J 22, 256 

Ascosterol, 343 

Asparagine, 300, 303, 309, 815 

D-Asparagine, 814 

L-Asparagine, 669, 791, 792 

Aspartate aminopherase, 488 

Aspartic acid, 290, 300, 301, 303, 
304, 308, 309, 311-313, 315, 
340-342, 424, 516, 531, 533, 
768, 769, 773, 813, 816-821, 
824, 826, 831, 836-839, 841, 
844, 845, 1078, 1079 

L-Aspartic acid, 514, 668, 814, 834 

Aspartic /?-semialdehyde, 311 

Aspartic transcarbamylase, 5i4 

Aspartocin, 445, 834 

^-Aspartyl phosphate, 311 

Aspelein, 1107 

Aspergillic acid, 497, 987, 988 

AspergilUn, 938 

Asperthecin, 547 

Asperxanthone, 890 

Astacin, 162 

Asterric acid, 191 

Aterrimin A, 1109 
B. 1109 

(ATP), adenosine triphosphate, 14, 
47, 53-55, 92, 93, 291, 311- 
313, 318, 333, 345, 425, 450, 
511, 514, 515, 524-526, 530, 
531, 533, 535-537, 556, 560- 
562, 564, 1040 



Subject Index 

(ATP), adenosine triphosphate 
-phosphoglyceric transphospho- 

rylase, 13 
-phosphopyruvic transphospho- 

rylase, 13 
synthesis, 449, 450 
Atranoric acid, 460 
Atranorin, 460, 857 
Atromentin, 235, 505 
-3,6-dibenzoate, 509 
Atrovenetin, 185, 570 
Aurantiacin, 509, 511 
Aurantiogliocladin, 236, 498, 512 
Aureomycin, 608 
Aureolic acid, 1110 
Aureothin, 870 

Aureothricin, 434, 870, 916, 1141 
Aurofusarin, 584, 888 
Auroglaucin, 107, 108, 189, 190, 

435 
Avenacein, 748 
Avidin, 423 
Ayfactin, J 22 
Ayfivin, 814 
Azafrin, 160 
Azalomycin B, 1111 

F, 1112 
Azaphilones, 879 
Azaserine, 532, 678 
Azomycin ( 2-nitroimidazole ) , 893, 

1197 

B-73, 304, 309 
Baccatine A, 1113 
Bacilipin A, 1114 

B, 1115 
Bacillomycin, 836 

A, 836 

B, 837 

C, 838 
R, 836 

Bacilysin, 1116 
Bacitracin, 343 

A, 814 

B, 814 

C, 814 

D, 814 

E, 814 

F, 814 



Pfizer Handbook of Microbial Metabolites 



720 



Bacitracin 

Fo, 814 

Fs, 814 

G, 814 
Bacterial carbohydrates, 338 

cell walls, 310, 314, 332, 343, 
344, 345, 422, 479, 514 

fats, 51 

pigments, 434 

polysaccharides, 528 

proteins, 345 

spores, 310, 314 
Bacteriochlorophyll a, 930 
Bacterioerythrin, 181 
Bacteriophage, 332, 344, 508, 509 
Bacteriopurpurin, 181 
Baeomycesic acid, 461 
Bamicetin, 1021 
Barbatic acid, 464, 861 
Barbatolic acid, 452 
Basidioquinone, 238 
Batatic acid, 854 
Behenic acid, 50 
Benzimidazole, 442, 446 
Benzoic acid, 618 
Benzoquinones, 185, 239 
Betaine, 683, 311, 466 
Biformin, 196 
Biformyne 1, 1117 
Binaphthyls, 214 
Biocytin, 426, 912 
Bioluminescence, 564 
Biomycin, 608 
Biopterin, 555, 1051 
Biotin, 54, 55, 92, 423, 424-426, 
447, 471, 526, 531, 532, 900 
Biotin- 1 -sulfoxide, 901 
Biphenyls, 2i4 
Bixin, 107 
Blasticidin A, 1118 

B, 1119 

C, 1120 
S, 1121 

Blastmycin, 185, 272 
B-Mycin, 760 
Boletol, 537 
Bongkrekic acid, 128 
Boninic acid, 483 
BorreUdin, 1122 



Bostrycoidin, 522 

Bottromycin, 760 

Brevin. 826, 827 

Brevolin, 827 

Bromogriseofulvin, 186, 431 

Bromotetracycline, 609 

Bryamycin, 840, 1292 

Bufotenin, 661 

2,3-Butanediol, 15, 19 

n-Butanol, 17, 18 

Butterfly wing pigment, 554, 1048 

iso-Butylamine, 466 

Butyryi coenzyme A, 54 

C-73, 291, 305, 295, 309 
Cadaverine, 466 
Caerulomycin, 1123 
Caldariomycin, 143, 144, 293 
Calycin, 630 
Camphomycin, 1124 
Candicidin A, 253 

B, 254 

C, 255 
Candicidins, 122, 253 
Candidin, J 22, 252 
CandiduUn, 1125 
Candimycin, 122 
Canescin, 1126 
Canthaxanthin, 163 

Caperatic acid, 49, 80, 81, 118, 159 

Caperin, 154 

Capraric acid, 451 

N-Carbamyl-L-aspartic acid, 514 

Carbamyl phosphate, 308, 514 

0-Carbamyl-D-serine, 671 

Carbomycin, 21, 119, 121, 283 
B,21, 119, 282 

Carbon dioxide, i4-J8, 47-49, 54, 
55, 92, 93, 292, 423, 424, 
447, 526, 527, 53 J, 536, 550, 
554, 558, 729, 731, 739 

l-Carboxy-2,5-dioxybenzyl methyl 
ketone, 402 

l-Carboxy-2,5-dioxyphenyl acetyl 
carbinol, i 85, 403 

Carboxylase, 13 

Carboxylation, 55 

4-Carboxy-2-oxo-3-phenyLhept-3- 
enedioic acid, 628 



721 



Subject Index 



3-Carboxy-2,4-pentadienal lactol, 

82, in 
N- ( 2-Carboxypheny 1 ) - 1 -aminori- 
bose-5-phosphate, 317, 459 
Carcinocidin, 848 
Carcinomycin, 847 
Cardelmvcin, 885 
Cardinophyllin, 1127 
Carimbose, 283 
Garlic acid, 148 
Carlosic acid, 79, 145, 149 
Carolic acid, 79, 146 
Carolinic acid, 147 
a-Carotene, 164 
^-Carotene, 91, 161, 162, 165, 185, 

186 
8-Carotene, 167 
y-Carotene, 94, 161, 166 
TT-Carotene, 176 
Carotene biogenesis, 90-94 
Carotenes, 90, 93, 107 
Carotenoids, 90, 94 
Carviolacin, 559 
Carviolin, 558 
Carzinophilin, 1127 

A, 1128 
Catenarin, 528, 541, 542, 546, 587 

6-methyl ether, 560 
Catenulin, 61 
Cathomycin, 885 
Cell tissues, 22 

walls, 22 
CDP-Choline, 1016 
Celesticetin, 120, 923 

I, 120, 258 
Celiomycin, 727 
Cellocidin, 5 
Cellulose, 512 
Cephalin, 1016 
Cephalins, 56, 135 
Cephalosporin C, 421. 367, 911 

N, 312, 367, 42i, 724, 905 

P, 368, 368 

Po, 369 

P3, 370 

P4, 371 
Cercosporin, 589 
Cerevioccidin, 1129 
Cerevlsterol, 344 



Cerinic acid, 124 
Cerotic acid, 124 
Cetyl alcohol, 47 
Chaetoalbin, 592 
Chaetochrysin, 590 
Chaetoflavin, 591 
Chaetomidin, 501 
Chanoclavine, 47J, 954 
Chartreusin, 439 
Chartreusin-like antibiotic, 440 
Chetomin, 941 
Chitin, 512 
Chitosamine, 33 
Chlamydosporin A, 1130 

B, 1131 
Chloramphenicol, 284, 342, 343, 

626 
Chlorine-containing peptide 

CssHaeO.N.Clo, 751 
Chloroatranorin, 459, 489 
7-Chloro-5a( 1 la)-dehydrotetra- 
cycline, 607 
7-Chloro-6-demethyltetracycline, 

602 
8-Chlorolevulinic acid, 143, 144 
Chloromycetin, 626 
Chlororaphine, 999 
Chlortetracycline, 608, 613 
Cholesterol, J 54 
Choline, 135, 311, 466, 554 
Choline phosphate, 56 
Chohne sulfate, 686 
Chromin, 122 
Chromomycin A3, 1132 
Chrysergonic acid, 535, 1133, 1152 
Chrysocetraric acid, 632 
Chrysomycin, 1134 
Chrysophanic acid, 539 
Chrysophanol, 538, 539, 592 
Cinerubin A, 617 

B, 617 
Cinerubins, 276, 606, 617 
Cinnabarin, 335, 502, 1001 
trans-Cinnamic acid, 620 

amide, 621 
Cinnamycin, 420, 816, 820, 821 
CircuHn A, 776 

B, 777 
Circulins, 776 



Pfizer Handbook of Microbial Metabolites 



722 



Citreorosein, 545 

Citric acid, 47, 48, 83, 95, 233, 466 
cycle, 46-49, 92, 93, 307, 309, 
445, 447, 561 
Citrinin, 411, 872 
Citromycetin, 185, 190, 410, 411, 

873 
Citrovorum factor, 1059 
Citrulline, 303, 308, 423 
Cladinose, 20, 278, 279 
Clavacin, 867 
Clavatin, 867 
Clavatol, 405 
Clavicepsin, 48, 466 
Claviformin, 867 
Clavine alkaloids, 470 
Clavorubin, 535 
Clavoxanthin, 553 
Cleavage enzyme, 53 
Clitocybin, 1135 
Cobalt, 445, 446 
Cobamic acid, 442 
Cobamide, 442, 444 
Cobamide coenzyme, 446 
Cobamide-containing polypeptides, 

444 
Cobamine cyanide, 931 
Cobinic acid, 442 
Cobyrinic acid, 441 

a,b,c,d,e,g-hexaamide, 442 
Cobyrinic acid pentamide, 442 
Cocarboxylase, 904 
Coccellic acid, 464 
Coelicolorin, 1136 _ 
Coenzyme A, 16, 47, 52, 53, 56, 
310, 527, 535, 556, 1046 
biosynthesis, 535-537 
Coenzyme 111 (nicotinamide ribose 
5'-diphosphate), 974 
Q^„ 237, 238, 512 
Q-, 237, 238, 513 
Q.^, 237, 238, 514 
Qg, 237, 238, 515 
Qio, 237, 238 
Coenzymes Q, 236-239, 247, 449, 

512 
Coliformin, 841 
Colimycin, 825 



Colistin, 771 
oi-Collatolic acid, 488 
Collinomycin, 1137 
Comenic acid, 406, 863 
Comirin, 824 
Compound A, 551, 1052 

CsHi40, 46 

C9H10O7N2 from Fusarium lyco- 
persici, 715 

CiiH.-.OgNo, 1138 

Ci.Ho^Oo, 46 

D, 393 

I, 823 

T, 376 
Condensing enzyme, 46, 93 
Congocidine, 918 
Coprinin, 493 
Coproporphyrin, 396, 437 

I, 927 

111, 438, 928 
Coproporphyrinogen, 438 
Cord factor, 52, 55, 139 
Cordycepic acid, 300 
Cordycepin, 2 J, 1032 
Cordycepose, 21 
Corphyrin, 445 
Corrin ring, 440 
Corticrocin, 219 
Cortisalin, 223 
Corynine, 137, 55 
Corynomycolenic acid, 131 
Corynomycolic acid, 54, 55, 121 

132 
Costaclavine, 952 
Cosynthetic factor-1, 1139 
Coupled phosphorylation, 449 
Cozymase 11, 904 
2, 6-Cresotic acid, 389 
Croceomycin, 1140 
Crocetin, J 07 
Crotonic acid, 160 
Cryptosterol, 352 
Cryptoxanthin, 171 
Cryptoxanthol, 171 
Crystallinic acid, 885 
Culmorin, 889 
Curvularin, 425 
Cyanocobalamin, 931 
Cyanomycin, 1141 



723 

Cycloheximide, 304, 307, 308, 

309, 310. 1228 
diastereoisomer, .'i09 
Cycloheximides, 144 
Cyclohexylamine salt, 1014 
Cyclopaldic acid, 409 
Cyclopenin, 493, 977, 981 
Cyclopolic acid, 411 
Cycloserine, 343, 345, 418, 422, 

488, 671, 894 
Cynodontin, 534, 544 
Cystathionine, 311, 420 
Cysteic acid, 300, 822 
Cysteine, 305, 310, 311, 340-342, 

422, 434, 447, 536, 718, 724, 

756, 757 
L-Cysteine, 333, 419, 420 
Cysteine-S-sulfonate,