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 I I I E I! Dl II [D 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 / 0 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- 0 II -C- 0 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 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° (-° (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.) [— 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 0 lo] *c \/\ * C CH— COOH *C COOH c -CO,> 0 *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 0 ±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°, [ 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, [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 0 0 CH3 1 II CH CH3 CH3 0 0 1 0 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 0 0 CH3 CH3 1 CH3 CH3 ^^ CH2 .^ OH 0 \ ^o^CVx ° 0 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). 0 OH / CH- -CH2— / NH \ 0 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 0 II CH- -CH 0 \ / Y NH y^ \^ v A HO CHs 0 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°, [■ 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 0 II 0 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— ^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 0 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^=( >=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 ) [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 -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 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 ® 0 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 0 +++ +++ + + + + + + + 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 .. j: o +++++++++++++ 1 + + + + + + ;; ^ o. 1 ■^ a 8 + + + + + + + E E "^ 3 3 O +++ + ++ + + + + + + + + ^• o .i.s -a u fe "o ^1 1^ " u 4> «> J° 4> u .= c c 0 a, c 0) 4) 4> £ c « ° a .S «> E :2 41 ■■= U C 4) c 0 5) .£ X 3- 0 t = ■£ 'E .E (u c c «> c c C c "0 £ •- i! 5 p - ° <0 acQ.?- 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 0 © "^\ 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= 0 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 © 0 (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). 0 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 0 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, 0 ^ 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 0 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 0 \ II — c — CH3 ^ / CH3 / 0 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\ 0 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 0 CH / ° L-Pro N / ^N -Q- CH2 / 11 / CH3 /. A 0 ^"^^^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 / 0 D-Hiv CH /^"^ _/ CH D-Val y^r. \ ^NH 1 0 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 Polypeptides and 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 Polypeptides 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 Polypeptides 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 0 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, 0 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- 0 0 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), [— 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 / 0 \ 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 0 0 / \/ 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 0 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'^ 0 // ~CH — / -NH^ V \„, 0 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 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\ 0 0 > I \ ^ ti II ^ Q c I I o^VSAch / \ / \ / \ CH3 CHs ,0" 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). 0 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 — 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 0 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 0 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 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 0 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 / ^^-^\ 0 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), [ [ 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 ^ 0 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.. _ 0 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 \ 0 CH- -CH2 T OH / CH3 \ .N^ < / -N^'\ HO i 0 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 0 T 0— P— 0— CH OH ^ Adenosine-3'-phospho- 5'-phosphosulfate 1 1 0 OH 1 HO — P— OH i 0 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 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- 0 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 0 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°, [ 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.), [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- 0 0 CH3 OH 0 -P— 0— P— 0— CH2-C CH— C— NH OH C > \ P(OH); i 0 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 0 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 0 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 0 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— 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-,> 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;-" —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). 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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 o ^ g CO 9 CD 1-^ I— I OO _ CO 'Vi CO o -Si O ^ o ^ 1-4 ^se P5 coi2 0) u o oq LO ,-1 CO eo CO ^ G CM JS •'1 CD H-i ;-; jr in « CT5 CD U rS « C3 © -c 00 oi « O rt 1/3 *J cq S o O O I' rr-l O 03 oq H C W cq Mo -S H I-H c^ c^J C/5 g 5 t5C/3 c- S CD O i-H >H o 'O rt ■" o J2 b O o g-g < o S ^^w'^is rt JH ffi^ w fl CI 2 ""Si t^«K^ W tf K fo ^ IIh- W d H ? ^.^ ^ ? W ^OeOt-OOOJHrH 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- 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^ — ^ !■ CN . — — CN >0 -* — — — 00 o lo — — — CN .— o ^— CN CO •— — 00 CO •— ^ o ^ ^ crT lo "O cn" ct" o O o :5 ^ •^ :2 rO CM lO — CN rv a) £ II U E o .= ■o 12 o o u — .- y o — 3 U 5 5 5 o o o '- 00 o nT - o , - ■<«■ . CN . ^ CO "o >o K CM >o -o ^ ~ — CO -o VO ■O — 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) 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 — -o r^v lO Is. — lO lO ^ "O <0 CN ^ lO — X o a 0) s CS 0 o> -Q c CN *u .o « o 0 > o> rs E CO < CN CN CN CN "o CN CN cs CN « CN ^ CN CN r— CN CN -^ CS o 0 CN CN 3 0 to -* -«t "* O o- o o CO ■* k' Is." -* rC -* - 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 o" 00 -a .£ -a _c ID 01 in Q u < a a 3 C 0 J3 a 2 '^ >. jj IT) S a c a o- c e- E 0 c E "^ o > c c D X c C c Ic 'c c c a Is c 2 u to Z c > ^3 2 i c 'c c C "e > j: 1- c N > 1- .. c 1- C a c < < c c -ri c Li .c X CL . '- CO CM lO •— O >- CN . >o" cn' O — — O- — CN COO ......... .— no>ootN-vO — -t^ooo — ■* coo — — CN-^OOOOOOOOO — .-•t — — — — — — — — CNfN CO >? ^ -= i o c 8 O CO CO . 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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. 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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. 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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. 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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 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-*■» 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 . 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 '^ 0 £ i J! 1 c O < < X .1 ^ ^ § - JC 4, J, ii (L 1 1 m ° - -- 1: £2 •c 2 o 1 ^ o < 0 *« •- « :i t ^- "I < "^^ o S S 5; llllh 1 - O < U < < Absorption X max. E.1% cm. o- CO >W CM CO o" CO <) 0 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| = ? 0 o I -E > O <^ 0) "S D ». -C 1= 8- m X o 1 ^ = "^ CO 3 8 -E 0) > o ^ 0) % D VI S- Uj X 0 i! ^ = 3 O w «> 0) 3 5 S 2 g • ui "E ? °' a i 3 £ ^ i s .. 1 § -s ;-. -5 S g 5 0 :i o „- "o -C a 0 .i: . 3 g . . d. E c a. i 0) J) £. 0 ■a o 4) C 1 D 0) o « 11. < E 0 u. CO « c "i o X 0 « u. 0) o Q. 0 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„ 0 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, 310 Cysteinylvaline, 421 Cystine, 303, 305, 420, 434, 722, 840, 848 L-Cvstinylvaline, 420 Cytidine, 509, 1010 Cytidine-5'-diphosphatecholine, 56. 512, 1016 Cytidinediphosphateethanol- amine, 512, 513 Cytidine diphosphate glycerol, 5i3, 5i4, 1015 Cytidine diphosphate ribitol. 5i3, 5J4, 1017 Cytidine nucleotides, 512, 513 Cytidine phosphate, 56 Cytidine-2'-phosphate, 1012 Cytidine-3'-phosphate, 1013 Cytidine-5'-monophosphate, 5i0 Cytidine-5'-triphosphate, 5J5 Cytidylic acid, 509, 1012, 1013 Cytidylic deaminase, 515 Cytochrome, 436, 447, 562, 564 aa, 449, 562 fla, 562 b, 449, 562 c, 447-449, 562 C4, 448 Cn, 448 Cytosine, 508, 509, 552, 1007 Subject Index Datemycin, 1142 Deca-tra7zs-2-irfln.s-8-diene-4,6- diyne-l,10-dioic acid, 199 Deca-cis-2-frflns-8-diene-4,6-diyn- l-ol, 204 Deca-fra7is-2-trans-8-diene-4,6- diynyl dec3i-trans-2-trans-8- diene-4,6-diynoate, 221 Decarboxylation, 309, 317, 422, 437, 447, 483, 492, 493, 502 2-Decene-l,10-dioic acid, 100 f rar2s-Dec-2-ene-4 ,6 ,8-triyn- 1 -al , 197 tra7zs-Dec-2-ene-4,6,8-triyn-l,10- diol, 201 7-Dechlorochlortetracycline, 273 Dechlorogeodin, 191 Dechlorogriseofulvin, 186, 432 Dechloronornidulin, 457 trarzs-a,/8-Dehydroacyl coenzyme A, 53 Dehydroaltenusin, 151, 418 Dehydrocarolic acid, 145 7-Dehydroergosterol, i54 14-Dehydroergosterol, 334 24(28)-Dehydroergosterol, 335 Dehydrofusaric acid, 479, 972 Dehydrogenase (DPNH), 449, 479-482, 561-564 5-Dehydroquinic acid, 143, 298 5-Dehydroshikimic acid, 143, 296 Dehydrotumulosic acid, 356 Dehydroustic acid, 393, 395, 412 6-Demethyltetracycline, 273, 603 Dendrolasin, 855 9-Deoxorosenonolactone, 329 3-Deoxy-3-amino-D-ribose, 21 6-Deoxy-L-gaIactose, 18 2'-Deoxy-5-methyloluridine-5'- phosphate, 552 Deoxyribonucleic acid (DNA), 345, 508-510 Deoxyribonucleotides, 5i5 Deoxyribose, 18, 445, 515 2-Deoxyribose-l -phosphate, 5i5 2-Deoxystreptamine, 20, 52, 53, 59 5-Deoxyoxy tetracycline, 273 Pfizer Handbook of Microbial Metabolites 724 2'-Deoxyuridine-5'-phosphate, 552 3'-Dephosphocoenzyme A, 537 Depsides, 212, 213, 231, 400, 402 Depsidones, 212, 213, 231, 400, 402 Depsipeptide, 1113 Dermocybin, 562 Desertomycin, 1143 Desosamine, 20, 120, 257, 258, 263, 276-279 Desthiobiotin, 902 Dethiobiotin, 426, 902 Dethiogliotoxin, 461 Detoxication, 232 Deuterium, 480, 481 Dextromycin, 62 Diacetyl, J 5, 19 Diadenosinetetraphosphate, 1045 a,/3-Diaminobutyric acid, 834 a,y-Diaminobutyric acid, 824 L-a,y-Diaminobutyric acid, 771, 776, 777, 779, 780-785 3,4-Diaminoguaiacol, 503 2,6-Diaminohexose, 20 Diaminohexose B, 60 a,e-Diamino-8-bydroxycaproic acid, 697 a,e-Diamino-^-hydroxypimelic acid, 717 2,6-Diaminopimelic acid, 306, 312-314, 343, 344, 703, 719 L,L-Diaminopimelic_acid, 312 meso-Diaminopimelic acid, 312, 343 a,/3-Diaminopropionic acid, 727 d-Diaminosuccinic acid, 305, 488, 670 4,5-Diaminouracil, 516, 557, 1008 Diaporthin, 1144 Diatretyne-1, 191 -2, 192 -3, 198 Diazoacetyl-L-serine, 678 6-Diazo-5-oxoaminohexanoic acid, 689, 725 6-Diazo-5-oxo-L-norleucine ( DON ) , 532, 689, 725 Dibenzofurans, 2 J 2, 214, 400 Dichloroacetic acid, 284 Dichloroproline, 739 Didymic acid, 212, 40 J, 861 Diffractaic acid, 467 D-Digitalose, 439 Digitoxigenin, 398 Diglyceride phosphate, 56 Dihydroagroclavine, 950, 952 Dihydroelymoclavine, 953 5,6-Dihydroergosterol, 341 Dihydro F, 909 Dihydrofuscin, 879 Dihydrogladiohc acid, 394, 410 DihydroUpoic acid, 16 Dihydronicotinic acid, 483 Dihydroorotase, 514 L-Dihydroorotic acid, 5i4 Dihydroorotic dehydrogenase, 514 Dihydrophenazine, 502 Dihydropyrazine, 497 Dihydroshikimic acid, 299 Dihydrostreptomycin, 19, 56 Dihydrostreptose, 19 4,5-Dihydrouracil, 516 Dihydroxy acetone, 16, 483 phosphate, 14 2,6-Dihydroxyacetophenone, 388 2,6-Dihydroxybenzoic acid, 185 3,4-Dihydroxybenzoic acid, 143 2,3-Dihydroxybenzoyl glycine, 396 2 ,6-Dihy droxybutyrophenone , 404 4-(D-er7/t/zro-l',2'-Dihydroxy-3'- phosphopropyl)imidazole, 318 a,/?-Dihydroxyisovaleric acid, 91, 315 3,3'-Dihydroxylycopene, 172 4 ,6-Dihy droxy-3-methoxyphtha- Uc acid, 393, 395, 412 a,^-Dihydroxy-^-methylvaleric acid, 97 725 4 ,9-Dihydroxyperylene-3 , 1 0-quin- one, 523 1,6-Dihydroxyphenazine, 995 3 ,4-Dihydroxyphenylalanine , 301 2,5-Dihvdroxyphenylglyoxylic acid, 384 3.5-Dihydroxyphthalic acid, 181, 185, 233, 386 3,5-Dihydroxy-l,4-pyrone, 72 2,5-Diketogluconic acid, 23, 25, 405, 406 a-Diketones, 422 Diketopiperazines, 346, 496, 497 2,3-Dimethoxy-5-methyl-l ,4-ben- zoquinone, 239 2,5-Dimethoxybenzoquinone, 494 1,8-Dimethoxynapthalene, 627 3-[2-( 3,5-Dimethyl-5-acetoxy-2- oxocy clohexyl ) -2-hy droxy- ethyl] glutarimide, 316 yS,/?-Diinethylacrylyl coenzyme A, 92 •y,y-Dimethylallyl pyrophosphate, 156 Dimethylamine, 291, 640 6-Dimethylaminopurine, 532, 534 trans-2,4-Dimethyl-13-7z-amyl-2- eicosenoic acid, 126 5 ,6-Dimethylbenzimidazole , 442, 444, 446, 529 a-(5,6-Dimethylbenzimidazolyl) cobamide cyanide, 441, 442, 931 Dimethyl deca.-trans-2-trans-8- diene-4 ,6-diyne- 1 , 1 0-dioate, 216 Dimethyl deca-2,4,6-triyne-l,10- dioate, 215 Dimethyl dec-trans-2-ene-4,6- diyne- 1,1 0-dioate, 217 Dimethylhistamine, 653 /3,5-Dimethyllanthionine, 420 L-bimethylleucine, 770 Dimethyl octa.-trans-2-trans-6- dien-4-yne-l,8-dioate, 206 3,5-Dimethyl-6-oxyphthalide, 400, 408 Subject Index Dimethylpyruvlc acid, 8 J, 86 6,7-Dimcthyl-8(D-l'-ribityl) lumazine, 557 Dimethylsulfone, 4 4,4'-Dioxo-/i-carotene, 163 2,6-Dioxy-5-methylpyrimidine, 509 2,6-Dioxypyrimidine, 508 2 ,4-Dioxy-6-pyru vylbenzoic acid, 185, 398 Dipalmitoleyl-a-lecithin, 136 D-l,3-Diphosphoglyceric acid, 14, 480 Diphosphopyridinenucleotide (DPN), 14, 47, 53, 93, 449, 479-481, 510, 511, 514, 527, 561-564, 975 Diphosphopyridine nucleic acid (reduced) (DPNH), 449, 511, 5J4, 561-564 2,6-Dipicohnic acid, 3 J 4, 479, 714, 968 Diplococcin, 1145 Diploicin, 441 Diploschisteric acid, 444 Dlpyrromethanes, 440 Dirhizonlc acid, 467 Disaccharldes, 511 Distamycin A, 1146 Divaricatic acid, 469 5,6,7,8,9,10,10',9',8',7',6',5VDo- decahydrolycopene, 178 Dodecyl 5-oxostearate, 12 Drosophihn A, 378 B, 1247 D-Substance, 1147 Duramycin, 420, 820 E-73, 305, 316 E-129A, 743 Eburicoic acid, 158, 355, 357 3/3-acetate, 357 Echinomycin, 760, 1089 Echinulin, 458, 460, 471, 496, 497, 943 Elaiomycin, 22, 711 Elaiophylin, 1148 Pfizer Handbook of Microbial Metabolites 726 Electron transport, 232, 233, 238, 447-450, 479, 561-564 Elymoclavine, 471,947 Embden-Meyerhof pathway, 13 Emodic acid, 536 Emodin, 188, 189, 234, 542, 562 Endocrocin, 154, 233 Endomycin A, 122, 235 B, 122, 246 Endosubtilysin, 1149 Endothianin, 580 Enniatin-A, 740, 747-750, 758, 767 Enniatin-B, 738, 747-750, 758, 767 Enniatin-C, 741, 747-750, 758, 767 Enolase, 13 Enolhydrase, 53 Enteromycin, 1150 Epanorin, 635 Episterol, 340 Epoxysuccinic acid, 49, 79 d,l-ETdm, 424 Ergobasine, 955, 956 Ergobasinine, 956 Ergochrysin, 1151, 1152 Ergoclinine, 955 Ergocornine, 465, 960, 961 Ergocorninine, 961 Ergocristine, 465, 966, 967 Ergocristinine, 967 Ergofiavine, 466, 1152 Ergokryptine, 465, 962 Ergokryptinine, 963 Ergometrine, 955 Ergonovine, 470, 955 Ergosecalinine, 957 - Ergosine, 958, 959 Ergosinine, 959 Ergostetrine, 955 A'"'-Ergosta-dien-3-one, 338, 346, 348 A^---Ergostadiene-3|8,5a,6;S-trioI, 344 A'--'-Ergostadien-3^-ol, 341 ^7.24(1:8) v-Ergostadien-3^-ol, 340 /^8,24(28) ?-Ergostadien-3^-ol, 342 A8.^^(?)-Ergostadien-3y8-ol, 343 5,7,22,24 ( 28 )-Ergostatetraene-3- /3-0I, 335 A'-Ergosten-3/3-ol, 345 Ergosterol, 154, 158, 336, 355, 466 biogenesis, i 55-158 peroxide, 339 Ergosteryl palmitate, 874 Ergot, 43, J 54, 291, 336, 343, 344, 351 alkaloid biosynthesis, 467-472 alkaloids, 291, 346, 458, 465, 472 Ergotamine, 470, 964, 965 Ergotaminine, 965 Ergothioneine, 319, 466, 708 Ergotocine, 955 Ergotrate, 955 Ergotoxine, 465, 470 Ergoxanthin, 1152 Erythrin, 468 Erythritol, 20, 468 weso-Erythritol, 20, 573 Erythrocin, 279 Erythroglaucin, 560 Erythromycin, 20, 119-121, 258, 279 B, 20, 119, 278 C, 20, 119,277 Erythronolide, J 20, 279 Erythropterin, 555, 1050 Erythrose, 398, 436 -4-phosphate, 17, 142 Erythroskyrin, 587 Escobedin, 160 Esperin, 763 Estin, 1153 Etamycin, 123, 382, 752, 769, 770 Ethanol, 14, 15, 18, 19, 466, 480 Ethanolamine, 135, 301 l-Ethoxy-l,2-ethylenedicarboxam- ide, 8 Ethyl acetate, 6 Ethylamine, 466, 639 Ethylcarlosic acid, 153 Ethylene, 3 f-tra Tis-Ethylene oxide-a,yS-dicar- boxylic acid, 79 Ethyl hydrogen 2,6-dipicolinate, 971 727 Subject Index Etiocobalamine, 442 Etrusconivcin, J 22, 228 Eulicin, 712, 1183 Eumycetin, 1154 Eumycin, 836 Eurocidin, i22, 242 Evernic acid. 446, 454 Exfoliatin, 1156 Expansine, 867 Factor A, 442 ribose phosphate, 442 Factor B, 932 C, 442 D, 442 E, 442 F, 442 G, 442 H, 442 I, 442 J, 442 K, 442 L, 442 M, 442 Via, 442 Vib, 442 Fairodin, 1157 Fallacinal, 548 Fallacinol, 557 Farcinicin, 914 Farinacic acid, 485 Farnesyl pyrophosphate, 156, 157 Fatty acid biogenesis, 52-54 catabolism, 53 ^-oxidation, 52 Fatty acids, 17, 22, 49, 50, 53, 93, 492 branched chain, 51 of microorganism fat, 50 of Mycobacterium tuberculosis, 51 Fatty alcohols, 23 Fecosterol, 342 Fermentation "lactobacillus casei" factor, 1061 Fermicidin, 1158 Fermizin, 1159 Fervenulin, 1160 Filipin, 119, 120, 122, 238 Flavacid, J 22, 244 Flavacol, 496, 497, 986 Flavensomycin, 1161 Flavicidin, 910 Flavicin, 910 Flavin, 47, 92, 449 Flavine-adenine dinucleotide (FAD), 479, 527, 560-564, 1060 Flavine biosynthesis, 557-560 enzymes, 56J Flavines, 548 Flaviolin, 516 Flavipin, 394 Flavofungin, 122, 225, 1143 Flavoglaucin, 436 Flavomycin, 1219 Flavoproteins, 561, 564 Flavoskyrin, 550 Flavucidin, 1162 Fluoride, 13 Folacin, 1058 Folic acid, 307, 346, 444, 531, 548, 549, 552, 554, 556, 1058 Folimycin, 1163 Folinic acid-SF, 1059 Fomecin A, 1164 Formaldehyde, 552 5-Formamido-4-imidazolecar- boxamide ribotide, 55i Formate, 445, 516, 550, 552, 554 Formate (Ci4-labeled), 120, 159, 182, 236, 411 Formic acid, 1 7, 46, 67, 72, 466, 552, 558 Formylglycinamide ribotide, 530, 551 N-Formylkynurenine, 482 N^"-Formylpteroic acid, 1055 6-Formylsalicylic acid, J 86, J 87 N'-Formyltetrahydrofolic acid, 1059 N^--Formyltetrahydrofolic acid, 549, 550 Forocidins, 289 Forocidin A, 289 B, 289 C, 289 Foromacidins, 119, 286-289 Pfizer Handbook of Microbial Metabolites 728 Fraction A (mitomycin), 1209 B (mitomycin), 1210 C (mitomycin), 1211 R (mitomycin), 1213 W-1 (mitomycin), 1206 W-2 (mitomycin), 1207 W-3 (mitomycin), 1208 Y (mitomycin), 1212 Fradicin, 122, 243 Fradiomycins, 60 Framycetin, 63 Frangula-emodin, 542 Frequentic acid, 873 Frequentin, 302 FriedeUn, 363 epi-Friedelinol, 364 Fructigenin, 749 Fructose, 19, 407 Fructose-1, 6-diphosphate, 14,17 Fructose-6-phosphate, 14, 17, 18 N-Fructosylanthranilic acid, 318, 459 D-Fucose, 18, 22, 439, 528 Fulvic acid, J 85, 186, 190, 410, 411, 875 Fulvicin, 430 Fumagillin, 107, 318 Fumarase, 46 Fumaric acid, 47-49, 78, 483, 531, 533 Fumaromono-D,L-alanide, 712 Fumarprotocetraric acid, 470 D,L-Fumarylyl alanine, 712 Fumidil, 318 Fumigacin, 367 Fumigatin, 495 Fumaric acid, 78, 309 Fumigatin hydroquinone, 496 Fungal cerebrins, 133 Fungichromatin, 122, 239 Fungichromin, 120, 122, 239 Fungicidin, J 22, 230, 237, 304, 305, 1095 Fungisporin, 778 Fungistatin, 839 Fungisterol, 345, 346, 348 Fungocin, 836 Funiculosin, 540 Furans, 398 Furan-3-carboxyIlc acid, 851 Furfural, 466 6-Furfurylaminopurine, 1030 Furoic acid, 417 Fusaric acid, 479, 973 Fusarium wilt toxin, 715 Fusarubin, 190, 410, 520, 521 Fusarubinogen, 521 Fuscin, 878 Fuscomycin, 1165 Galactonic acid, 32 3-;8-D-Galactopyranosido-D-arabi- tol, 39 Galactose, 22, 471 Galacturonic acid, 22 Gallic acid, 187, 382 Gangaleoidin, 449 G-Compound, 557-560, 1053 Geamine, 732, 734, 735, 773 Gentian violet, 343 Gentisaldehyde, J 87 Gentisic acid, 186, 187, 381 Gentisyl alcohol, 186, 187, 383 Gentisylquinone, 491 Geodin, 191, 213, 424 d-Geodin, 426 Geodin-like antibiotic, 429 Geodoxin, 427 Geomycin, 729, 734, 735, 773 Gibberellenic acid, 321, 322 Gibberellic acid, 323 biosynthesis, 159 GibberelUns, 321, 479 Gibberellin A, 321, 325 Ao, 321, 326 A3, 321, 323 A4, 324 X, 321, 323 Gigantic acid, 909 Glabratic acid, 443 Gladiohc acid, 408 Glauconic acid I, 317 11, 317 Glauconic acids, 144, 317 GUorosein, 499 Gliotoxin, 145, 346, 458, 460, 461, 496-498, 938 acetate, 939 729 Gliotoxin biosynthesis, 460, 461 Glomellifeiic acid, 181 D-Gluconic acid, J 8, M, 77, 405 Gluconolactone, 405 5-0-a-D-Glucopyranosyl-D-fructo- pyranosc, 41 2-0-a-D-Glucopyranosyl-D-glucose, 42 D-Glucosamine, 18, 20, 23, 33, 59, 344 Glucosamine isomer, 732 6-Glucosamine, 19, 52 Glucose, 14, 17, 18 l-C'*-Glucose, 182, 555 Glucose-6-phosphate, 14, 17, 480, 511, 524 Glucosides, 5i2 a-D-Glucosido-a-D-glucoside, 43 Glucosone, 18, 24, 407 D-Glucuronic acid, 18, 28, 82, 109 Glucuronides, 511, 512 ^rfl7is-Glutaconic acid, 84 Glutamate, 319, 435, 480, 515, 530, 533 Glutamate-aspartate aminophe- rase, 487 Glutamic acid, 274, 290, 300, 301, 303, 304, 306-309, 312, 333, 340-342, 344, 436, 445, 501, 516, 530, 533, 549, 730, 768, 773, 813, 815, 819-821, 828, 829, 831, 836-838, 841, 848, 849, 1062, 1078 D-Glutamic acid, 343, 814 L-Glutamic acid, 92, 333, 680, 718, 722, 730, 765, 766 Glutamic acid semialdehyde, 307 Glutamine, 300, 303, 308, 318, 515, 530, 532 L-Glutamine, 681, 791, 792 L-y-Glutamylcysteine, 333 Glutamylcysteinylglycine, 718 Glutaric acid, 88 Glutathione, 307, 310, 311, 332, 333, 340, 420, 480, 718, 722 L-Glutathione, 333 Glutathione-cysteine disulfide, 722 Glutinosin, 1166 Subject Index D-Glyceraldehyde-3-phosphate, 1 4, 17, 18, 480 L-(-)-Glyceric acid, 46, 76, 483 Glycerin, 17 Glycerol, 17, 55, 483, 501 Glycine, 300, 301, 303, 305, 309, 310, 333, 340-342, 419, 434- 437, 444, 516, 530, 550, 552, 554, 558, 663, 718, 722, 752, 754, 759, 760, 766, 768, 769, 773, 790, 815-818, 820, 822, 824, 826, 828, 829, 831, 834, 840, 841, 846, 848, 1079, 1222 Glycine-2-C^\ 435 Glycinamide ribotide, 530, 55J Glycogen, 5J2 Glycolic acid, 72 Glycolipide from Pseudomonas aeruginosa, 130 Glycolysis route, 13, 14, 561 Glycylcystine, 435 Glycylglycine, 436 Glyoxylate, 3i0 Glyoxylic acid, 48, 49, 87, 3 JO, 550 cycle, 48 Gramicidin, 1145 Gramicidins, 786 Gramicidin C, 788 D, 789 J:, 787 J2, 786 S, 339, 340, 788 Dubos, 789 Gram-negative bacteria, 3J4 Granatacin, 576 Granegillin, 988 Grasseriomycin, 736 Grifolin, 46 Grisamine, 1167 Grisein, 765, 766 Griseoflavin, 1168 Griseofulvin, 186, 188, 189, 213, 411, 430, 431 Griseolutein A, 502, 1004 B, 1005 Griseomycin, 265 Griseoviridin, 1169 Pfizer Handbook of Microbial Metabolites 730 Grisovin, 430 Grizein, 843 Guanidine, 2 Guanine, 22, 442, 508, 509, 529, 559, 1027, 1044 Guanine-S-C^*, 558 Guanosine, 509, 1034 diphosphate, 527, 533 diphosphate factor B, 442, 528, 529 diphosphate fucose, 528 diphosphate mannose, 511, 527, 528 nucleotide, 527 Guanosine-3'-phosphate, 1039 Guanosine-5^-monophosphate (GMP), 5iO Guanosine-5^-triphosphate (GTP), 511, 527, 533 Guanyhc acid, 533, 1039 L-Gulonolactone, 82 D-Gulosamine, 21, 731, 729 Gyrophoric acid, 475, 476 Haematommic acid, 2i3 Haemocorin, 573 Helenine, 1170 Hehomycin, 1171 Hehxins, 247 Helixin A, J 22, 235 B, J 22, 246 Helminthosporin, J 89, 541 Helvolic acid, 367, 368 Hematin, 925 Heme, 436 proteins, 447, 561' Hemin-hke substance, 46 Hemipyocyanine, 994 Hemoglobin, 436 2-n-Heptyl-4-oxyquinoUne, 978 2-n-Heptyl-3-oxy-4-quinolone, 492, 979 2-n-Heptyl-4-oxyquinoHne N-oxide, 980 Hercynine, 319, 707 Herquein, 572 Herqueinone, 571, 573 Heteroxan thine, 1028 Hexacosanoic acid, 124 9-Hexadecenoic acid, 50, 106 Hexokinase, 13, 524 Hexose phosphate, 18 rz-Hexylamine, 466, 647 Hiascic acid, 476 Hiochic acid, 96 Hirsutic acid, 1172 Histamine, 466, 651 Histidine, 300, 301, 303, 305, 318, 319, 341, 342, 448, 466, 816- 819, 849 L-Histidine, 688, 814 Histidine betaine, 707 biosynthesis, 3i8, 551 L-Histidinol, 3 J 9, 691 phosphate, 319 Holomycin, 434, 435, 913 Homocysteine, 311, 419, 420, 553 Homarine, 701 Homogentisic acid, 143 Homomycin, 21, 58 amino sugar moiety, 21, 58 Homoprotocatechuic acid, 384, 387, 391 Homosekikaic acid, 478 Homoserine, 311, 312, 315 deaminase-cystathionase, 487 isomerase, 485 Hyaluronic acid, 5i2 Hydrogen transport, 232, 447— 450, 479-481, 561-564 yS-Hydronapthazin, 521 a-Hydroxy acids, 564 L-^-Hydroxyacyl coenzyme A, 53 ^-Hydroxyacyldehydrogenase, 53 3-Hydroxyanthranilic acid, 482, 483 5-HydroxyanthraniHc acid, 460 Hydroxyaspartic acid, 488 Hydroxyaspergillic acid, 989 5-Hydroxybenzimidazole, 442 p-Hydroxybenzoic acid, J 43, 186, 187, 379 3-Hydroxy-y-carotene, 170 (,j-Hydroxycatenarin, 549 10-Hydroxydec-trans-2-ene-4,6-di- ynoic acid, 205 731 Subject Index trans-10-Hydroxydec-2-ene-4,6,8- triynoic acid, 198 (,)-Hydroxycniodin, 515 5-Hydroxyindole, 470, 471 D(-)a-Hydroxyisovaleric acid, 338, 339, 738, 740, 747-750, 758, 767 a-Hydroxyketones, 422 3-Hydroxykynurenine, 482 3/^-Hydroxylanosta-8,24-diene-21- oic acid, -MO 21-Hydroxylanosta-7,9( 11 )24-tri- ene-3-one, 346 /^-Hydroxyleucine, 726 2- ( 6-Hydroxy-2-methoxy-3 ,4-meth- ylenedioxyphenyl ) -benzof u- ran, 858 1-Hydroxydimethoxymethylxan- thone, 890 a-Hydroxymethyl-a'- ( N-acetylami- nomethylene ) succinic acid, 485 6-Hydroxy-2-methylaminopurine, 532 6-Hydroxy-2-methylbenzoic acid, 389 5-Hydroxy-2-methylchromone, 868 5-Hydroxymethylcytosine, 509, 5J5, 552 5-Hydroxymethylfuran-2-carbox- ylic acid, 852 5-Hydroxymethylfurfural, 398 ;8-Hydroxy-/?-methylglutaryl co- enzyme, A (HMG-Co A), 92, 93, 155 cleavage enzyme, 93 8-Hydroxy-3-methylisocoumarin, 397 /3-Hydroxy-^-methyl-8-valerolac- tone, 96 D-^-Hydroxymyristic acid, 104 Hydroxymycin, 64 Hydroxy-P-481, 180 1-Hydroxyphenazine, 994 p-Hydroxyphenylacetic acid, 390 p-Hydroxyphenyllactic acid, 316 p-Hydroxyphenylpyruvic acid, 87, 235, 316 3-Hydroxyphthalic acid, 186, 187, 385 3-Hydroxypicolinic acid, 770 Hydroxyproline, 436. 756, 757, 769, 805, 807, 820 D-a-Hydroxyproline, 770 Hydroxypyruvic acid, 87 Hydroxyspirilloxanthin, 181 Hydroxystreptomycin, 19, 55 Hydroxystreptose, 19 3-Hydroxy-5-toluic acid, ,389 5-Hydroxytryptamine, 935 5-Hydroxy tryptophan, 470, 471 ^-Hydroxyvaline, 419 Hygromycin A, 21, 57, 185 B, 21, 45 Hygroscopin A, 1173 B, 1174 Hygrostatin, 1112, 1175 Hyposterol, 332 Hypothamnolic acid, 463 Hypoxanthine, 442, 532, 1023 Illudin M, 1176 S, 1177 Ilotycin, 279 Imbricaric acid, 473 Imidazole, 418 Imidazoleacetol, 687 phosphate, 319 Imidazoleglycerol, 690 phosphate, 3J8, 319 4-Imidazoly acetic acid, 677 Imoticidin, 1178 Inactone, 307 Incrassatic acid, 861 Indigo, 458, 940 Indigoidine, 1179 Indole, 306, 317, 318, 471, 933 ^-Indoleacetic acid, 417 Indole-3-acetic acid, 934 Indole biosynthesis, 3 J 7, 459 synthetase, 488 Indoles, 458, 496 3-Indolylacetol, 460 Indolyl-3-gIycerol phosphate, 3 J 7, 459 Inosine, 529 nucleotide, 527 Pfizer Handbook of Microbial Metabolites 732 Inosme-5'-phosphate, 510, 1035 Inosinic acid, 532, 533, 1035 meso-Inositol, 30 Inositols, 19 lodinin, 996 lodoacetate, 13 Ipomeamarone, 855 Ipomeanine, 853 Iridoskyrin, 540, 583, 587 Iron, 765, 766 Islandicin, 540, 587 Islanditoxin, 739 Isoamylamine, 291, 652 Isobutyl acetate, 9 Isobutylamine, 648 Isocitric acid, 47, 48 allo-Isocitric acid, 93 Isocitric dehydrogenase, 46 Isocitritase, 48 Isokojic acid, 404, 407, 866 Isoleucine, 80, 97, 300-302, 304, 314, 315, 342, 497, 815-818, 824, 831, 839, 840 L-Isoleucine, 693, 776, 777, 814 D-a^o-Isoleucine, 336, 793, 795- 801, 804 Isoleucine biosynthesis, 315 Isolysergic acid, 466 Isooosporein, 502 Isopenniclavine, 949 Isopentenyl pyrophosphate, i55, J 56 Isoprene, 9, 470, 471 Isopropanol, 18 Isopropylamine, 645 ' Isopyridoxal, 484 Isorhodomycin A, 276, 597, 1180 B, 598 e-Isorhodomycinone, 276, 601 Isorhodomycinones, 276 Isosetoclavine, 946 Isovaleryl coenzyme A, 92 mycarose, 283 Isoxazole, 418 Itaconic acid, 49, 83 Itaconitin, 1181 Itatartaric acid, 89 Javanicin, 235, 520 Junipal, 427, 895 Kanamycin, 19, 20, 52 B, 53 Kanosamine, 20, 53 Keto-acids, 87, 422 /?-Ketoacyl coenzyme A, 53 a-Ketoacyldehydrogenase, 92 a-Ketoadipic acid, 312 ^-Ketoadipic acid, i43, 144 a-Ketobutyric acid, 315 a-Ketocaproic acid, 87 2-Keto-3-deoxy-D-araboheptonic acid, J 42 5-Keto-6-deoxyarabohexose, 21 2-Keto-3-deoxy-6-phosphogluconic acid, 18 L-2-Ketofucopyranose, 21 2-Ketogalactonic acid, 27 2-Keto-D-gluconic acid, 25, 72, 405 5-Keto-D-gluconic acid, 26, 72, 405 a-Ketoglutaraldehyde, 550 a-Ketoglutarate, 319, 480 a-Ketoglutaric acid, 47, 48, 85, 87, 92, 284, 307, 309, 312, 437, 527 2-Keto-L-gulonolactone, 82 a-Keto-^-hydroxyisovaleric acid, 3J5 a-Ketoisocaproic acid, 87, 92, 316 a-Ketoisovaleric acid, 87, 3 J 5, 316, 333 5-Ketostearic acid, 112 Kinetin, 534, 1030 Kojibiose, 42 Kojic acid, 404, 405, 407, 865, 866 Krebs cycle, 46, 47 Kynurenine, 336, 337, 482, 493 Lactams, 79 Lactarazulene, 320 Lactarinic acid, 112 Lactaroviolin, 319 Lactic acid, 17, 46, 80, 466, 717 d-Lactic acid, 15, 75 L-Lactic acid, J 5 L( + )-Lactic acid, 75, 338, 339 Lactobacillic acid, 51, 114 733 Subject Index Lactobionic acid, 44 Lactones, 79 Lactonic acid. 1239 Lagosin. i 79. i 20, 229 Lankaniycin. 119, 120 A^'-*-Lanostadien-3-ol, 352 A'-^'^'-'^-Lanastatriene-S/^, 21-diol, 348 Lanosterol, 157-159, 352 Lanthionine, 420, 815-821 Laricic acid, 120 Lateritiin-I, 740, 758, 767 Lateritiin-II, 747, 758. 767 Laterosporin, 1182 Latumcidin, 1183 Laurie acid, 104 Lavendulin, 774 Lecanoral, 488 Lecanoric acid, 443, 444, 468 Lecanorolic acid, 488 Lecithin. 1016 a-Lecithin. 136 /^-Lecithin, 136 Lecithin biosynthesis, 513 Lecithins, 55, 135 Lenamycin, 1184 Lenzitin, 1185 Leprapic acid, 633 Leprapinic acid, 633 Leprotene, 188 Leprotin, 188 Leucine, 91, 92, 300, 301, 304, 309, 314-316, 339, 341, 342, 466, 497, 759, 768, 784, 813, 815-818, 824, 828, 829, 831, 837, 838, 841, 849, 1078 D-Leucine. 771, 776, 777, 780, 785-787, 790, 825 L-Leucine, 339, 501,692, 726, 771, 781, 782, 788, 791, 792, 814 Leucine biosynthesis, 314-316 Leucomelone, 236, 506 Leucomycin, 275 Leucopterin, 554 Leucotylin, 366 Leucovorin, 1059 Leucrose, 41 a-Leucyl-L-leucine anhydride, 1195 a-Leucyl-a-proline anhydride, 1194 Levomycetin, 626 Levomycin, 753 Lichen acids, 284 Licheniformin A, 844 B, 845 C, 846 Z-Lichesterinic acid, 80, 156, 159 Lichexanthone, 891 Lignoceric acid, 50, 122 Limocrocin, 224 Linoleic acid, 50, 51 Linolenic acid, 50 Lipoic acid, 1 5- J 7, 47, 99 Lipoproteins, 50 Liposaccharides, 50, 52 Litmocidin, 1186, 1305 Lobaric acid, 480 Lomycin, 265 Longisporin, 1187 Lusomycin, 161 Lustericin, 1188 Lutein, 174 Luteol, 174 Luteoleersin, 578 Luteomycin, 577 Luteoskyrin, 587, 588 Lycomarasmine, 715 Lycopene, 94, 161, 168 Lycopersene, 94 Lycopersin, 1189 Lycophyll, 172 Lysergic acid, 466-471 Lysine, 300, 301, 303, 305, 307, 312, 313, 341-344, 426, 815- 818, 824, 831, 839, 841, 844, 845, 848 ^-Lysine, 727, 729, 731, 732, 734, 735, 773, 790 L-Lysine, 306, 343, 695, 814 Lysine biosynthesis, 312, 313, 314 Lysozyme, 332, 343, 344 D-Lyxuronic acid, 21 M5-18903, 1064 Macrocyclic lactones, 118, 122 Macrolide antibiotics, 118, 190 Macrosporin, 556 Magnamycin, 119, 283 Pfizer Handbook of Microbial Metabolites 734 Malate synthetase, 48 L-Malic acid, 47, 48, 81, 483 Malic dehydrogenase, 46 Malonic acid, 71 Malonyl coenzyme A, 54, 155, 424, 446 Maltobionic acid, 44 Maltose, 44 Malucidin, 1190 Mannan, 528 D-Mannitol, 19, 35, 43, 46, 71, 329, 407, 466, 874 D-Mannonic acid, 32 D-Mannopyranosyl-l-Tneso- erythritol, 38 Mannose, 22 Mannosidostreptomycin, 65 Marasin, 200 Marasmic acid, 1191 Marcescin, 922 Marcomycin, 1192 Matamycin, 822 trans, trans-Matricaria acid, 202 ester, 211 trans,trans-Matricarianol, 203 Mycocidin, 367 Mediocidin, 245, J 22 Megacidin, 1193 Melanomycin, 849 Melanosporin, 1196 Mellein, 399 Mesaconic acid, 445 Mesenterin, 1197 Mesoinositol monophosphate, 34 MetaboUte Co4H5o02,_1198 Metabohte A, 628 B, 628 from Curvularia lunata, 1200 of Coprinus comatis, 1199 of Eremothecium ashbyii, 1008 of Hydniim aurantiaciim, 511 Metal chelates, 436 Metamycin, 1201 Methionine, J 89, 291, 300-304, 310, 311, 337, 420, 444, 445, 461, 516, 525, 552-554, 798, 816-818 L-Methionine, 684 Methionine (C^Mabeled-CHg), 120, 159, 274, 411 6-Methoxybenzoxazolidone, 896 8-Methoxy-l-naphthol, 613, 627 p-Methoxyphenylalanine, 535 p-Methoxytetrachlorophenol, 378 4-Methoxytoluquinone, 493 2-MethyIadenine, 442, 534 S-Methyl-S-adenosylmethionine , 292 Methylamine, 291, 466, 638 Methylaminoethanol, 646 6-Methylaminopurine, 532 Methyl anisate, 622 a-[L],^-Methylaspartic acid, 445, 516, 834 C-Methylation, 236 O-Methylation, 236 5-MethylbenzimidazoIe, 442 2-Methyl-2-butene, 7 a-Methylbutyric acid, 90 Methyl trans-cinnamate, 623 Methyl p-coumarate, 624 /3-Methylcrotonyl coenzyme A, 424 S-Methyl-L-cysteine, 305, 676 5-Methylcytosine, 509 Methyl 10-(deca-trarzs-2,trans-8- diene-4 , 6-diyn- 1 - oyloxy ) -dec- trans-2-ene-4,6-diynoate, 222 Methyl-2,4-dideoxy-2-aminotetro- side, 22 N'^,N'^'-Methylenetetrahydrofolic acid, 549 N-Methyl-L-glucosamine, 19, 54 /?-Methylglutaconase, 92 trans-^-Methylglutaconic acid, 81, 94 ^-Methylglutaconyl carboxylase, 92 coenzyme A, 92, 424 1-Methylguanine, 532 2-Methyl-2-heptene-6-one, 10 Methyl 1 0-hydroxydec-trans-2-ene- 4,6-diynoate, 212 Methyl tra?is-10-hydroxydec-2-ene- 4,6,8-triyn-l-oate, 210 4-Methyl-5-(2-hydroxyethyl)-thia- zole, 422 2-Methylhypoxanthine, 442 735 Subject Index N-Methvlisoleucine, 337, 796, 797 /J-Methvllanthionine, 305, 704, 8 15-82 1 N-Methylleucine, 741 Methylmalonyl coenzyme A, 447 2-Methylniercaptoadenlne. 442 Methyl p-methoxycinnamate, 284, 625 Methyl 2-methoxypulvinate, 633 2-Methyl-l,4-naphthoquinone, 239 6-Methyl-l,4-naphthoquinone, 517 1-10-Methyloctadecanoic acid, 115 ( + )-6-Methyloctanoic acid, 338, 771, 776, 777, 780, 781, 783- 785 S-Methylol-S-adenosylhomocy- steine, 553 5-Methyl-2-oxo-4-imidazoUdine- caproic acid, 902 3-Methyl-3-oxyglutaryl coenzyme A, 155 6-Methyl-7-oxy-8-(D-Li-ribityl)- lumazine, 557 y-Methylproline, 726 6-MethylsalicyIic acid, 185-189, 233, 236, 389 y-Methyltetronic acid, 80, 140 4-Methylthiazole, 422 Des-N-methylthiolutin, 913 3-Methyluracil, 765 5-Methyluracil, 515 N-Methyl-D-valine, 337 N-Methyl-L-vaHne, 738, 740, 747- 750. 793-812 Methymycin, 20. 119, 121, 261 Mevaldic acid, 93, 155 2-C^*-Mevalonic acid, 159, 160 Mevalonic acid lactone, 81, 91, 93, 96, 119, 154-156, 189, 190, 239, 398 Mevalonic acid dipyrophosphate, 156 5-monophosphate, 156 3-phosphate 5-pyrophosphate, 156 5-pyrophosphate, 156 Miamycin, 120, 292 Microcin A, 1202 B, 1203 Micrococcin, 761 Micrococcins, 761 Micrococcin-P, 762 Micromonosporin, 1203 Microphyllic acid, 212, 489 Mikamycin, 770 A, 1204 B, 1205 Mineoluteic acid, 49, 105 Mitochondria, 236, 238 Mitomycin C, 1214 Mitomycins, 1206-1213 Mitoquinone, 247, 511 Moldin, 1215 MoUisin, 235, 519 Monascin, 879, 882 Monascorubrin, 879, 884 Monilin, 1104, 1105, 1216 Monoacetylprotocetraric acid, 465 Mucopeptides, 343 Mucopolysaccharides, 22 Muramic acid, 343-345 Musarin, 1112, 1217 Muscaridine, 659 Muscarine, 291, 658 Muscarufin, 508 Muscle adenylic acid, 1038 Mushroom poisons, 458 Mutomycin, 1218 Myacins, 60 Mycaminose, 21, 290 Mycarose, 21, 290 Mycelianamide, 186, 411, 497, 998 MyceUn, 1219 IMO, 1220 Mycifradin, 60 Mycobacidin, 899 Mycobacillin, 813 Mycobactin, 185, 772 Mycoceranic acid, 124, 129 phthioceryl ester, 129 Mycocerosic acid, 129 Mycochrysone, 525 Mycoin, 867 Mycolic acid, 51, 55, 138 Mycolipenic acid, 125 Mycolutein, 634 Mycomycin, 218 Pfizer Handbook of Microbial Metabolites 736 Mycophenolic acid, 185, 186, 188, 189, 433 Mycorhodin, 1221 Mycosamine, 20 Mycose, 43 Mycospocidin, 1222 Mycostatin, 230 Mycosubtilin, 842 Mycothricin, 1223 A, 734 B, 735 Mycoticin, 1224 Mycoxanthin, 189 Myoinositol, 82 Myprozine, 226 Myristic acid, 50, 103, 104 triglyceride, 103 Nalgiolaxin, 566 Nalgiovensin, 567 Naphthoquinone from Mycobac- terium phlei, 530 Naphthoquinones, J 85, 235, 248 Naramycin A, 308 B,310 Narbomycin, 20, 119, 121, 274 Natural penicillins, 905 Neamine, 60, 61, 63 Nebularine, 1031 Necrosamine, 662 Nemotin, 208, 209 Nemotinic acid, 108, 209 xyloside, 108, 109, 220 Nemoxynic acid, 478 Neobiosamine C, 60_ NeohydroxyaspergiUic acid, 990 Neoinosamine-2, 21 Neomethymycin, 20, 119, 262 Neomins, 60 Neomycin, 20, 63, 64 Neomycins, 60 Neomycin A, 60 B, 60, 62 C, 60 Neophromin, 561 Neosamine C, 20, 60 Neospiramycins, 289 Nephromopsic acid, 80, 81, 159 Nephrosteranic acid, 154, 155 Nephrosterinic acid, 154 Netropsin, 346, 435, 918 Neuraminic acid, 344 Neuraminopeptides, 344 Neurosporaxanthin, 187 Neurosporene, 94, 175 Neutral nitrogen-containing com- pound, 877 Ngaione, 855 Nicotinamide, 480, 563 ribose 5'-diphosphate, 974 Nicotine, 435 Nicotinic acid, 479, 483, 974, 1054 biosynthesis, 482, 483 Nidulin, 212, 466 Nigericin, 1225 Nisins, 420, 816 Nisin A, 816 B, 817 C, 818 D, 819 Nitrogen-containing compound, 876 2-Nitroimidazole, 893 p-Nitrophenylserinol, 284 ^-Nitropropionic acid, 73, 310 Nitrosporin, 257 Nivemycins, 60 Nocardamin, 713 Nocardianin, 1226 Nocardorubin, 1227 Noformicin, 730, 915 Nonactin, 1228 ( — )-Nona-3,4-diene-6,8-diyne- l-ol, 200 trans-Non-2-ene-4,6,8-triyn-l-al, 193 f rans-Non-2-ene-4 ,6 , 8-triyn- 1 -ol, 194 (2d,3d)-Nona-4,6,8-triyn-l,2,3- triol, 195 2-(n-A'-Nonenyl)-4-oxyquinoline, 982 2-7i-Nonyl-4-oxyquinoline, 983 N-oxide, 984 Nordin, 1153, 1229 Norherqueinone, 571, 573 Nornidulin, 456 737 Norstictic acid, 447 Norvaline, 755 Notatin, 850 Novaose. 21 Novobiocin, 21, 343, 885 NTCC 7197, 1116 Nucleic acids, 508, 524 Nucleocidin, 1043 Nucleoproteins, 508, 5J0 Nucleosides, 509 Nucleotides, 509 Nudic acid A, 1230 B, 192 Nybomvcin, 1231 Nystatin, 20, 122, 230 Obtusatic acid, 454 Ochracin, 399 Ochrolechaic acid, 442 Octacosan, 11 d-2-Octadecanol, 50 d-3-Octadecanol, 51 7.8,ll,12,12Ml',8',7'-Octahydro- lycopene, 177 Octapyrrole, 440 Odyssic acid, 108, 209, 214 Odyssin, 209, 213 Oleandomycin, 20, 119, 121, 276 Oleandrin, 118, 119 Oleandrose. 20, 119, 276 Oleic acid, 50, 51 Oligomycin A, 1232 B, 1233 C, 12.34 Olivetoric acid, 212, 486 Ommatins, 1001 Ommochromes, 335 One-electron transfer, 446 Oosporein, 501 Ophiobalin, 1235 Oregonensin, 1236 Orientomycin, 894 Ornithine, 300, 307, 308, 312, 423, 436, 820, 1078 D-Ornithine, 786, 787, 814 L-Omithine, 339, 666, 685, 786- 788, 791, 792 Orosomycin, 1295 Subject Index Orotic acid, 52 5 riboside, 1014 Orotidine, 1014 Orotidine-5'-phosphate, 515 Orotidylic decarboxylase, 515 pyrophosphorylase, 515 Orsellinic acid, 81, 186, 190, 213, 233, 392, 401 4-methyl ether, 81 Orygmaeic acid, 504 Oryzacidin, 1237 Oryzasizine, 1237 Ostreogrycin A, 743, 770 Oxalic acid, 49, 68, 466 Oxaloacetic acid, 47, 48, 80, 81, 87, 308, 309, 423, 483, 527 Oxalosuccinic acid, 47 decarboxylase, 46 Oxamycin, 422, 894 Oxidase (cytochrome a^), 449 Oxidative phosphorylation, 524 L-4-OxopipecoHc acid, 755 Oxoproline, 802, 803 2-Oxy-6-aminopyrimidine, 508 3-Oxyanthranilic acid, 502, 1054 3-Oxy-^-carotene, 171 4-Oxy-/3-carotene, 171 Oxychlororaphine, 501, 998 (-)-3-Oxydecanoic acid, 338, 723, 759 2-Oxy-5-hydroxymethyl-6-amino- pyrimidine, 509 Oxyjavanicin, 521 i-3-Oxykynurenine, 337, 1054 3 a-Oxylanost a-8 , 24-diene-2 1 -oic acid, 350 methyl ester-acetate, 349 S-Oxy-L-lysine, 697 2-Oxy-5-methyl-6-aminopyrimi- dine, 509 3-Oxy-4-methyl-anthranilic acid, 336, 337 3-Oxypalmitate, 80 4-Oxyquinolines, 492, 493 Oxytetracychne, 273-275, 306, 610, 670, 1302 Oxytetracycline-X, 274, 275 Pfizer Handbook of Microbial Metabolites 738 P-481, 179 PA 94, 894 105, 276 106, 1076 107, 1076 108, 280 114A, 743, 754, 770 114B, 744, 755, 770 114B-3, 745, 770 121, 614 128, 1238 132, 1239 133A, 260 133B, 264 147, 141 148, 281 150, 122, 250 153, 122, 240 166, 122, 227 Pachybasin, 538 Palitantin, 144, 302, 303 cis-Palmitenone, 13 Palmitic acid, 50, 51, 54, 55, 104, 108, 121, 124 Palmitoleic acid, 106 Palmitone, 14 Panmycin, 613 Pannaric acid, 859 Pannarin, 453 Pantetheine, 334, 536, 720 Pantetheine-4'-phosphate, 536 Pantethine, 721 Pantoic acid, 333, 535 Pantothenic acid, 333, 334, 536, 537 d-Pantothenic acid, 716 Pantothenic acid 4'-phosphate, 536 ■Pantothenylcysteine, 334, 536 Paraconic acid, 485 Parellic acid, 442, 450 Parietin, 2J2, 555 Parietinic acid, 554 ParmeUn, 460 Paromamine, 59 Paromobiosamine, 59 Paromomycin, 20, 59, 61 Paromose, 20, 59 Patulin, 186, 188, 867 biosynthesis, 82 Penatin, 850 ^-Penetrin, 417 Penicidin, 867 Penicillamine, 419, 420, 911 Penicilhc acid, 81, 144 biosynthesis, 81 Penicillin, 3ii, 337, 343, 345, 346, 716, 970 B,850 F, 910 G, 906 K, 907 X, 908 Penicillins (biosynthesis), 418- 421,435 Penicilliopsin, 585 Penitrinic acid, 423 Penniclavine, 948, 949 Pentacosanoic acid, 123 Pentaenes, 119 Pentamycin, i22, 241 Pentose, 405 phosphate, 18 oxidative cycle, 13, 17, 18 Peptolide, 1113 Perlatolic acid, 482 Peroxidase, 436 Perylenequinones, 235 Phagolessin A 58, 1240 Phalamycin, 1241 Phalofacin, 1242 Phalloidin, 458, 756 Phalloin, 757 Phenanthrenequinone, 233, 234 Phenazine, 502, 997 Phenazine-1-carboxamide, 999 Phenazine- 1-carboxylic acid, 997 l,6-Phenazinediol-5,10-dioxide, 996 Phenazines, 501 1-Phenazinol, 994, 1000 Phenol coupling, 191, 213, 214, 234, 400-402, 502 Phenohc substances, 185, 212, 213, 236, 502 Phenoxazones, 50J, 502 Phenylacetic acid, 417 739 Phenylalanine, 143, 182, 300, 301, 303, 305, 306, 316, 342, 461, 470, 497, 815. 820, 821, 849 D-Phenvlalanine, 339. 778, 779, 781-783, 786-788, 791, 792, 814 L-Phenylalanine. 705, 755, 778, 787, 791 Phenylalanine biosynthesis, 316 /:J-Phenyl-/^-alanine, 739 ytf-Phenyl-^-aminopropionic acid, 751 /3-Phenylethylamine, 291, 466, 656 Phenylpyruvic acid, 143, 235, 284, '316. 493 Phleomycin, 1243 Phloroglucinol, 188 Phoenicin, 500 Phomazarin, 569 Phosphate, 18, 56, 450, 480, 524, 530, 531, 533, 560, 562-564 Phosphatides, 51, 52 Phosphoenolpyruvic acid, 14, 142, 527 6-Phosphogluconic acid, 17, 18 D-2-Phosphoglyceric acid, 14, 77 D-3-Phosphoglyceric acid, 14, 310 Phosphoglyceromutase, 13 Phosphohexoisomerase, 13 Phosphohexokinase, 13 2-Phospho-4-hydroxy-4-car- boxyadipic acid, 98 3-Phosphohydroxypyruvic acid, 310 Phospholipide biosynthesis, 52 Phosphohpides, 50 Phosphoribose pyrophosphokinase, 524 5-Phosphoribosyl-l-pyrophos- phate, 317, 515 Phosphoric acid, 14, 17, 333 Phosphorylase, 485, 487 Phosphoserine, 310 Phosphotidylethanolamine, 513 Photosynthesis, 436, 564 Phthienoic acid-Coy, 126 Phthiocerol, 66 Phthiocerol dimycoceranate, 66 Phthiocol, 518 Subject Index Phthioic acid, 51, 124 Phthiomycin, 728 Physarosterol, 353 Physcion, 2] 2, 555, 560, 573 anthranols, 563, 564 Physetohc acid, 106 Physodalic acid, 465 Physodic acid, 485 Phytoene, 94, 177 Phytofluene, 178 Phytomonic acid, 114 Phytonivein, 1244 Picoline, 752 Picrocin, 20, 263 Picrolichenic acid, 437 Picromycin, 20, 119, 121, 263 Picrorocelhn, 497, 992 Pigment I, 417 II, 417 A, 1002 Pigment B (fois-anthraquinone), 581 (phenazine), 1003 C, 582 R, 182 Y, 183 Pimaricin, 20, 119, 121, 122, 226 Pimelic acid, 426 Pinastric acid, 632 PinicoUc acid A, 347 Pipecohc acid, 338 D-a-Pipecohc acid, 314, 834 Piricularin, 1245 Pleocidin, 733, 734, 735 Pleomycin, 1246 Pleuromutihn, 1247 Pleurotin, 1248 Phcacetin, 1020 Pluramycin A, 1249 B, 1250 Poin, 1251 Polyacetylenes, 109 Polycycline, 613 Polyene macrolides, i20 Polyenes, J 07 cts-Polyisoprene, 9 Polymyxin, 671, 780 A, 780 Bi, 781 Pfizer Handbook of Microbial Metabolites 740 Polymyxin B.,, 782 C, 783 D, 784 E, 785 Polypeptide antibiotics, 332 biosynthesis, 332, 345, 346 Polypeptides, 299, 332, 508, 511 (intracellular), 332, 340-342 Polypeptin, 779 Polyporenic acid A, 359 C, 354 Polyporic acid, 235, 504 Polysaccharide, 922 Polysaccharides, 22, 511 Polystictin, 1001 Porphobilinogen, 437-440 deaminase, 439, 440 Porphyrilic acid, 857 Porphyrin byosynthesis, 436-440 enzymes, 447-450, 561 Porphyrinogens, 440 Post-oxidation, 236 Prephenic acid, 143, 301, 316, 493 Primycin, 1252 Proactinomycins, 266 Proactinomycin A, 266 B, 267 C, 268 Prodigiosin, 435, 436, 919 precursor, 435, 436, 920 Prodigiosin-like pigment, 436, 924 Porphyrins, 310, 434, 444, 447, 448 Proline, 300, 301, 303, 304, 307, 342, 435, 436, 796, 797, 799, 801, 803-807, 813, 815-818, 820, 821, 831, 837, 839, 844, 845, 849, 1078 L-Proline, 336, 339, 679, 754, 755, 766, 786-788, 791-795, 812, 834 L-ProUne (C^Mabeled), 435 1,2,3-Propanetriol, 17 Propionate, i20, 447 Propionic acid, 46, 74, 447 acid-l-C^*-H3, 120 Propionyl coenzyme A, 424, 447 2-Propionylthiazole-4-carboxylic acid, 762 Propiopyrro thine, 916 zso-Propylamine, 466 7i-Propylamine, 466, 644 Propynoic acid, 108 Protein biosynthesis, 332, 343, 345, 346, 534 Protoactinorhodin, 527 Protocarbomycin, 121 Protocatechuic acid, 380, 382 Protocetraric acid, 119, 213, 451 Protocidin, 122, 232 Protoleucomelone, 510 d-Protolichesterinic acid, 157 Z-ProtoHchesterinic acid, 157, 159 Z-afio-Protolichesterinic acid, 158 Protomycin, 1314 Porphyria, 438 Protoporphyrin, 437, 926 IX, 438, 447 Protoporphyrinogen, 438 Psalhotin, 1253 Pseudoneamine, 64 Pseudopsoromic acid, 455 Psicofuranine, 1042 Psilocin, 936 Psilocybin, 458, 936, 937 Psoromic acid, 450 Pteridine, 554, 558, 1063 biosynthesis, 555-558 pigment, 1063 Pteridines, 548 Pterin-hke substance, 1049 Pteroproteins, 549 Pteroylglutamic acid, 554, 1058 Pteroyl-y-glutamyl-y-glutamyl- glutamic acid, 1061 Pteroylhexaglutamylglutamic acid, 1062 Puberulic acid, J 82, 373 Puberulonic acid, i82, 183, 375 Pulcherrimin, 991 Pulcherriminic acid, 496, 497, 991 Pulvic anhydride, 2i2, 629 Pulvilloric acid, 1254 Pulvinic acid, 235, 236 Pumilin, 1255 741 Purine biosynthesis, 424, 530-533, 558 nucleoside, 559, 560 nucleotides, 529, 559 Purines, 308, 310, 318, 422, 508, 524-538, 557-559, 564 Puromycin, 21, 534, 535, 1047 Purpurogenone, J 90, 411, 874 Putrescine, 291, 292, 466, (>50 Pyo compounds, 492 Pyocyanine, 50i, 1000 Pyolipic acid, 107 Pyoluteorin. J 85, 435, 917 Pyrans, 404-407 Pyrazines. 496 PyTidomycin, 752, 770 Pyridoxal-5-phosphate, 92, 310, 312, 437, 484, 487, 969 Pyridoxamine, 484 phosphate, 484 5-Pyridoxic acid, 484 Pyridoxine, 310, 479, 484-487, 970 phosphate, 484 Pyridoxol, 484 Pyrimidine biosynthesis, 514 nucleotides, 509, 510 Pyrimidines, 309, 508 Pyrocalciferol, 337 Pyroclavine, 951 Pyrogallol, J 86, 187, 377 Pyrogens, 52 Pyrones, 404, 405, 407 Pyrophosphate, 53, 333, 511, 515, 524, 526, 533, 560 Pyrroles, 434, 458 gi.pyrroline-5-carboxylic acid, 307, 435 Pyrromycin, 606 7/-Pyrromycin, 615 e-Pyrromycinone, 606, 616, 617 ^-Pyrromycinone, 605 r/-Pyrromycinone, 604 Pyrromycinones, 275, 276 Pyruvate, J5-i7, 80, 81, 309, 315, 560 Pyruvic acid, 14, 18, 46-48, 70, 80, 87, 309, 312, 313, 315, 316, 423, 559 Subject Index Q,.7,, 247. 511 Quadrilineatin, 401 Quinhydrones, 232 Quinic acid, 143 Quinocyclines, 275, 276, 614 Quinolines, 492 Quinones, 23 J, 449 Quinonoid compounds, 4J0 Racemomycin A, 790, 1256 B, 790, 1257 C, 790, 1258 Ractinomycin A, 1259 B, 1260 Radicalisin, 586 Radicinin, 413, 871 Raisnomycin, 1261 Ramalic acid, 454 Ramalinic acid, 451 Ramalinolic acid, 474, 471 Rammacin, 1262 Ramycin, 1263 Rangiformic acid, 49, 117 Raromycin, 1264 Roseomycin, 1265 Ravenelin, 886 Resistomycin, 575 Resorcinol, i88 Respiratory chain, 447-450, 561— 564 Reticulin, 55 Rhamnose, 723 Rhizobacidin, 1266 Rhizocarpic acid, 636 Rhizoic acid, 464 Rhizopin, 934 Rhizopterin, 1055 Rhodocidin, 1267 Rhodocladonic acid, 212, 565 Rhodomycetin, 529, 1271, 1305 Rhodomycin, 22, 275, 276 A, 276, 596, 1180 B, 276, 598 /3-Rhodomycinone, 276, 599 y-Rhodomycinone, 276 e-Rhodomycinone, 276, 600 Rhodomycinones, 275, 276 Rhodophyscin, 595 Rhodopin, 169 Pfizer Handbook of Microbial Metabolites 742 Rhodopurpurene, 168 Rhodosamine, 22, 276, 615 Rhodovibrin, 180 Rhodoviolascin, 184 8-Ribityl-6 , 7-dimethyllumazine , 1053 8-Ribityl-6-methyl-7-oxylumazine, 1052 Riboflavin, 516, 529, 555, 1056 biosynthesis, 557-560 Riboflavin-5'-phosphate, 560, 1057 9- ( ^-D-Ribof uranosyl ) purine , 1031 Ribonucleic acid (RNA), 345, 508-510, 526, 532, 534 Ribonucleoprotein , 1170 D-Ribose, 18, 59, 60, 317, 318, 483, 560 Ribose-5-phosphate, 17, 318, 458, 524, 530 Ribose-5-phosphate-l-pyrophos- phate, 524, 530 5-Ribosyluracil, 509 Ribulose-5-phosphate, 17 Rifomycin B, 593 Rimocidin, 122, 231, 1095 Ristocetin A, 1268 B, 1269 Roccellic acid, 49, 50, 110 Rosenonolactone, 160, 328, 330 biosynthesis, 159, 160 Roseonine, 731, 732, 734, 735, 790 Roseopurpurin, 558 Roseothricin A, 21, 729, 732 Rosololactone, i59, J 60, 330 Rotaventin, 1270 Rotiorin, 879, 883 " Rubidin, 1305 Rubiginic acid, 72, 406, 864 Rubiginol, 72, 406, 862 Rubixanthin, 170 Rubrofu sarin, 887 nor-Rubrofusarin, 890 Rubroglaucin, 560 Rubromycin, 1271 Rubropunctatin, 880 Rubroskyrin, 587 Rugulosin, 580, 586 Ruticin, 1272 Rutilantinone, 276, 606 SA, 247, 511 Saccharic acid, 29 Sacromycin, 1022 Salazinic acid, 448 Salmotin, 905 Sambucinin, 750 Sarcidin, 1273 Sarcinaxanthin, 186 Sarcinene, 186 Sarcolactic acid, 75 Sarcosine, 310, 337, 664, 770, 793-812 Sarkomycin, 294 Saxatilic acid, 448 Sclererythrin, 1152 Sclerocristallin, 1152 Sclerotiorin, 411, 881, 883 Scleroxanthin, 1152 Scopularic acid, 455 Secaclavine, 954 Secalonic acid, 1152, 1274 Sedoheptulose-1 ,7-diphosphate, 142 Sedoheptulose-7-phosphate, 1 7 Sekikaic acid, 454, 471 Seligocidin, 1275 Senecioyl coenzyme A, 92 Serine, 300, 301, 303, 304, 309- 311, 341, 342, 419, 435, 461, 471, 497, 516, 552, 739, 759, 773, 813, 816, 822, 824, 826, 836, 839, 841 D-Serine, 422, 671, 766, 784 L-Serine, 317, 459, 667, 723, 727, 734, 735, 751 Serotonin, 458, 935 Serratamic acid, 723 Setoclavine, 945, 946 Shikimic acid, 143, 297, 3J7 biosynthetic route, J 42, 143, 181, 188, 236, 284, 316, 317, 458, 493 5-phosphate, 3i6 Sinanomycin, 918 Sirenin, 1276 Sistomycosin, 122, 234 Skyrin, 234, 580, 582, 587 SLR Factor, 1055 Soframycin, 63 743 Solanorubin, 168 Solorinic acid, 574 Sorbicillin, 107, 117, 423 Spurassol, -10() Spermidine, 291, 292, G55 Spermine, 291, 292, 660 Sphaerophoric acid, 462 Sphaerophorin, 472 Spheroidenone, 182 Spheromycin, 885 Spiculisporic acid, 49, 50, 105, 109 Spinulosin, 145, 497 Spiramycins. 21, 119, 286-288 Spiramycin I, 286 II, 287 III, 288 Spirilloxanthin, 184 Sporidesmin, 1277 Squalene, 154, 157-159, 351 Squamatic acid, 461, 462, 861 Stachydrine, 702 Staphylomycin M, 742, 754, 770 M.., 755, 770 S, 755, 770 Stearic acid, 50, 51, 113 Stearyl alcohol, 49 Stereocaulic acid, 480 Sterigmatocystin, 892 Steroid glycoside, 118 Steroids, 154, 157 Sterol esters, 50 Sterols, 46, 93, 158, 160 Stictaic acid, 455 Stictic acid, 455 Stictaurin, 2J2 Stipitatic acid, 182, 372 Stipitatonic acid, 182, 183, 374 Strepogenins, 333 Strepsilin, 856 Streptidine, 19, 54 Streptimidone, 315 Streptobiosamine, 54 Streptocardin, 1278 Streptogramin, 746, 770, 832 Streptolidine, 729, 731 Streptolin A, 729 Streptolins, 729 Streptolydigin, 1279 Streptomycete antibiotics, 19 Subject Index Streptomycin, 19, 54, 63-65 B, 65 Streptomycins, 19 Streptonivicin, 885 Streptose, 19, 54 Streptothricin, 21, 729, 731, 732- 737, 790 Streptothricin BI, 60 BII, 60 Streptothricins, 60 Streptovaricin A, 1280 B, 1281 C, 1282 Streptovitacin A, 311 B, 312 Co, 313 D, 314 Streptozotacin, 1285 Strophanthin, 118 Substance 1404, 1286 Subtihn, 420, 815, 816 Succinate, 17, 447, 483 Succinic acid, 47, 48, 80, 313, 466, 550 Succinic dehydrogenase, 46, 449, 561 Succinyl coenzyme A, 47, 424, 437, 446, 447, 527 transferase, 93 N-Succinyl-L-diaminopimelic acid , 719 N-Succinyl-L-glutamic acid, 714 Sucrose, 24 Sugar nucleotides, 22 Sugars from streptomycete anti- biotics, 19 Sulcatic acid, 450 Sulfocidin, 1288 Sulfactin, 1287 Sulfanilamide, 531 Sulfate, 310, 524, 525 Sulfokinase, 525 Sulfur, 310, 427, 461, 498 Sulfur bacteria, 930 Sulochrin, J 9 J, 428 Sumiki's acid, 398, 852 Suprasterol, 154 Synnematin B, 312, 421, 724, 905 Pfizer Handbook of Microbial Metabolites 744 T 1384, 918 Tabtoxinin, 717 Taitomycin, 1291 D-Talose, 21, 45 Taraxerene, 362 Tardin, 1292 L( + )-Tartaric acid, 82 Tartronic acid, 72, 80 Taurine, 301 Telomycin, 769, 770 Teloschistin, 557 Tennecetin, 122, 236 Tenuazonic acid, 80, 151 Tenuiorin, 484 Teropterin, 1061 Terpene biosynthetic route, 81, 159, 160, 239 Terphenylquinones, 235, 236 Terramycin, 274, 610 Terramycin-X, 274, 612 Terrecin, 1293 Terreic acid, 492 Terrein, 295 Terrestric acid, 152 Tertiomycin A, 284 B, 285 Tetracosanoic acid, 122 Tetracycline, J 85, 190, 273, 613, 1139 biosynthesis, 273-275 Tetracyclines, 273 Tetracyn, 613 2,3,4,6-Tetradeoxy-4-dimethyla- minohexopyranose, 21 Tetrahydrofolic acid, 310, 311, 333, 515, 530 6,7,6',7'-Tetrahydrolycopene, 175 Tetrahydronicotinic acid, 483 Tetrahydroxybehenic acid, 121 Tetrahydroxybenzoquinone, 490 4,5,4',5'-Tetrahydroxy-l,l'-dina- phthyl, 524 l,4,7,8-Tetrahydroxy-2-methyl-an- thraquinone, 551 3,4,3',4'-Tetraoxo-y3-carotene, 162 Tetraphenylhydrazine, 502 Tetrapyrrole, 440 Tetronic acid, 79 acids, 79, 398 Tetrose phosphate, 18 Thamnolic acid, 121 Thelephoric acid, 507, 511 Thermophillin, 568 Thiactin, 1294 Thiamine, 418, 422, 423, 560, 903 Thiamine diphosphate, 904 Thiamine pyrophosphate, 15, 16, 47, 92, 315 Thiazole, 418 /?-(2-Thiazole)-/:?-alanine, 760 Thiazolidine-4-carboxylic acid, 422 Thioaurin, 1294, 1295 6,8-Thioctic acid, 99 Thiolutin, 434, 914 Thiomycin, 1296 Thiophene, 418 Thiostrepton, 831 Thiosulfate, 310 Thiourea, 1 D-Threitol, 20 Threonine, 300-302, 303, 304, 305, 310-312, 315, 341, 342, 444, 752, 755-757, 759, 762, 769, 773, 831, 836, 839, 840, 1079 L-Threonine, 675, 771, 776, 777, 780-785, 793-812, 825 Threonine synthetase, 485 Thymidine diphosphate rhamnose, 1019 Thymidine-5'-phosphate, 552 Thymine, 422, 445, 509, 515, 516, 529, 552 Tiglic acid, 417 Tobacco mosaic virus, 5iO a-Tocopherol, 438 D-L-Tocopherol, 239 Torularhodin, 161, 185 Torulene, 161, 185 Totomycin, 1297 Toxin of Helminthosporium victo- riae, 768 Toxin of tobacco wild-fire disease, 717 Toxoflavin, 1029 Toyokamycin, 1104, 1105 745 Toyocamycin, 1298 Trametenolic acid, -MO Transaminase, 92, 485 Transamination, 290, 291, 485, 486, 493, 550 Transhydrogenase (TPN-DPN), 449 Transmethylation, 311 Transpropionation, 447 Trehalosamine, 20, 40 Trehalose, 43 Trehalose phosphate, 511 Tricarboxylic acid cycle, 46, 47 Trichomycin, J 22, 251 Trichothecin, 160, 327 biosynthesis, 159, 160 Triglycerides, 50 Trigonelline, 700 2 ,4 ,5-Trihydroxyphenylglyoxylic acid, 387 Trimethylamine, 291, 466, 643 ( + )-2,4L,6L-Trimethyltetracos-2- enoic acid, 125 Triose, 405 Triose phosphate, 14, 18, 317, 458, 459 Triosephosphate dehydrogenase, 13 isomerase, 13 Triphosphopyridinenucleolide (TPN, Codehydrase II), 17, 47, 54, 93, 449, 479, 480, 510, 550, 976 Triphosphopyridine nucleotide (re- duced) (TPNN), 315, 449, 528, 553 Triseclavine, 946 Triterpenes, 93, 154, 157 Tritisporin, 549 Tropolone acids, 181 biosynthesis, 181-183 Tryptophan, 143, 299, 301, 302, 305, 306, 317-319, 336, 337, 342, 458, 470, 482, 483, 493, 756, 757, 789, 815, 831, 839 ^-C'*-Tryptophan, 470, 471 L-Tryptophan, 459, 460, 471, 709, 790, 792 Tryptophanase, 306 Subject Index Tryptophan biosynthesis, 317, 318 synthetase, 485 Tubercidin, 1299 Tuberculin, 342 Tuberculostearic acid, 115, J22, 124 Tubermycin A, 997 Tumulosic acid, 356, 358 Tylosin, J J 9, 290 Tyramine, 466, 657 Tyrocidine, 1145 A, 791 B, 792 Tyrocidines, 786 Tyrosine, J 43, J 82, 305, 316, 341, 342, 466, 497, 813, 824, 826, 836-839 D-Tyrosine, 1078 L-Tyrosine, 706, 791, 792 ?neta-Tyrosine, 461 Tyrosine biosynthesis, 316 Tyrothricin, 786, 787, 789, 791, 792 Ubiquinone, 247, 511 UDPG, 1018 Umbilicaric acid, 479 Umbilicin, 39 Unclassified compound, 1300 Undec-3,5,6-triene-8,10-diynoic acid, 207 10-Undecenoic acid, 101, 102 2-n-Undecyl-4-oxyquinoline N- Oxide, 985 10-Undecylenic acid, 102 10-Undecynoic acid, 101 Ungulinic acid, 119 Unnamed antibiotic, 1301 Unsaturated Co acids, 50 Uracil, 22, 508, 529, 552, 1006, 1044 Urea, 308 cycle, 308 Ureidosuccinic acid, 5J4 Uric acid, 1025 Uridine, 1009 diphosphate, 510-512 Uridinediphosphateacetylglucosa- mine, 5J0, 5J2, 1018 Pfizer Handbook of Microbial Metabolites 746 Uridinediphosphate-L-arabinose, 511 Uridinediphosphategalactose, 510, 511 Uridinediphosphateglucose, 520- 512, 1018 Uridinediphosphateglucuronic acid, 511 Uridinediphosphate-D-xylose, 511 Uridine nucleotides, 343, 510-512 Uridine-3'-phosphate, 1011 Uridine-5'-phosphate(UMP), 510, 515 Uridine-5'-pyrophosphate, 343 Uridine-5'-triphosphate, 510-512, 515 Uridylic acid, 1011 Urocanic acid, 305 Uroporpliyrin III, 438, 440, 929 Uroporphyrinogen, 438, 439 III, 440 isomerase, 439, 440 Uroporphyrins, 438 UrsoHc acid, 361 Usnarin, 460 Usnetic acid, 480 Usnic acid, 159, 212, 400, 401, 454 d-Usnic acid, 460, 860 /-Usnic acid, 462, 857, 860 Ustic acid, 393, 395, 412 Ustilagic acids, 127 Ustilic acid A, 127 B, 127 Ustin, 456 II, 457 cis- Vaccenic acid, 51, 111 Valine, 91, 301, 303, 304, 309, 314-316, 340-342, 466, 497, 724, 759, 768, 798, 799, 802- 810, 815-818, 820, 821, 828, 829, 831, 838, 839, 841, 845, 846, 849, 1078 D-Vahne. 337-339, 419, 758, 767, 778, 790, 793-795, 811, 812 L-Valine, 337-339, 419, 682, 758, 760, 778, 786-788, 790-792, 834 D-Valine-1-C^*, 338 L-Valine-1-C^*, 338, 420 Valine biogenetic pathway, 81, 314, 315 Valinomycin, 123, 337-339, 747- 750, 758, 767 Vancomycin, 1302 Variolaric acid, 442 Variotin, 1303 V-Compound, 557, 560, 1052 Vengicide, 1304 Ventosic acid, 121 Verdazulene, 321 Versicolorin, 543 Vertimycin C, 1305 Vicanicin, 445 Victoxinine, 768 Vinacetin, 594 Vinactane, 727 Vinactin A, 727 Viocin, 727 Violacein, 458, 942 Violacetin, 1306 Violarin, 1307 Viomycin, 727, 729, 734, 735 Viridicatic acid, 153 Viridicatin, 493, 977, 981 a-Viridin, 1308 yS-Viridin, 1309 Viridogrisein, 770 Virtosin, 1310 Viruses, 508 Viscosin, 723, 759 Vitamin A, 239 Vitamin B, 1058 conjugate, 346, 1062 Vitamin Bi, 903 diphosphate, 904 Vitamin B.., 1056 Be, 970 Bjo, 311, 434, 436, 440-444, 446, 447, 516, 528, 529, 552, 554, 931 pseudo-Vitamin Bj^, 442, 445 Vitamin Bj., analogues, 442 Vitamin C,"l43 D.., J 54 D3, J 54 E, 438 747 Vitamin C H, 423 K, 237-239, 512 K...A, 5:n K.,,B. 5:{2 K...C. 5.-{:{ K,„ 531 d-Volemitol, '56 Volucrisporin, 503 Vulcamycin, 885 Vulpinic acid, 631 Waksman's actinomycin B, 12 Watermelon wilt toxin, 1244 Wortmannin, 1311 Xanthicin, 1312 Xanthine, 532, 559, 1024 Xanthocillin-X, 284, 434 Xanthocillin-Y, 434 Xanthommatin, 235, 336, 1001 Subject Index Xanthomycin4ike antibiotic, Xanthomycins, 1314 Xanthones, 185 XanthophvU, 174 Xanthopterin, 554, 556, 1018 Xanthothricin, 1315 Xanthylic acid, 532, 533 Xyhndein, 528 L-Xylose, 88, 109 Xylulose-5-phosphate, 17 Yeast adenylic acid, 1037 cerebrin, 134 Zaomycin, 219, 835 Zeaxanthin, 173 Zeaxanthol, 173 Zeorin, i 57, .36.5, 635 Zymonic acid, 80, 142 Zymosterol, 7 59, 331 1313 EMPIRICAL FORMULA INDEX This index lists the known empirical formulas of microbial metabolites as an aid to future characterizations. Boldfaced numbers are entry numbers, while italic numbers are page numbers reflecting occurrence in a chapter or section introduc- tion. The appendixes and addendum are not indexed. NHg, 637 C,H704N, 668 CHoOo, 67 C^HsO.., 6 CH4N0S, 1 C4H^03N.., 669 CH5N, 638 C4HSO4N.., 670, 671 CH5N3, 2 C4HSO4N4, 672 C0H0O4, 68 C4H9O0N, 673, 674 C0H4, 3 C4H9O3N, 675 C0H4O2, 69 C4H9O3NS, 676 C0H5O0N, 663 C4H106, 18 CoHeO, 15 C4H10O0, 19 CoHeOoS, 4 C4H10O4, 20 C.,H,N, 639, 640 C4H11N, 648 C0H7ON, 641 C4H11ON, 649 C3H3O0N3, 893 C4H10N0, 650 C3H4O3, 70 C-,H40-., 1068 C3H4O4, 71 C5H4ON4, 1023 C3H40,„72 C-,H402N4, 1024 C3H,,04N, 73 C;H403, 851 C h'C N^'894 C,H403N4, 1025 C3He03, 16, 75 ^•^JJ^^^' ^f C,U,0„ 76 C5H5N5, 1026 C3H7ON, 642 C5H5ON5, 1027 CgH.OoN, 664, 665, 666 CgHeOoNs, 677 C3H7O3N, 667 C5H6O3, 140 C3H,07P, 77 - C5H6O4, 83, 84 C3H8O3, 17 CgHeOg, 85 C3H9N, 643, 644, 645 C5H7O4N3, 678 C3H9ON, 646 CgHgOo, 417 C4H4O2N,,, 5, 1006, 1184 CHgOoClo, 293 C4H4O3N0, 1184 CgHsOa, 86 C4Hr,ON3, 1007 . C5HSO4, 88 C4H4O4, 78 C,HsOe, 89 C4H4O,, 79 C5H.,N3, 651 C4H6O2N4, 1008 C,H,OoCa/2 • 2H..0, 21 C4H,,04, 80 C.H^O.N, 679 C4H6O5, 81 C,H904N, 680 C4He06, 82 C5H10, 7 749 Empirical Formula Index C-,H,„0.., 90, 417 C,H,„0,N.., 681 C,H,„0^. 91 C-,H„O..N, (i82. 683 C,H,,O..NS, 681 C-,H,..O..N.., 685 C,H,.0-„ 22 C,H,,N, 652 C-,H,.ANS, 686 C.jH^b-,, 863 C.-H^O^;, 490. 864 C,.H-,O..N,, 1048 C,,H„O..N4, 1028, 1029 C^-H.jO^, 141, 786,377 C,;H,.,Oj, 852, 865, 866 C,.,H,;0-„ 142 C,jH,.0,„ 92, 93 C^HsO-.N., • HCl, 687 C,;Hs04, 94 CfiH.O,,, 143 CoHsO,, 95 CgHgOoN,, 688 CC.H9O3N3, 689 (CcHioOo)n, 1148 CeHioO.,, 96 CeH,„0,N,, 8 CfiHioOaNo • HCl, 690 CgHioOe, 24 CeHioO^, 25, 26, 27, 28 CeHi„0«, 29 CeHuON;, • HCl, 691 CgHnO^N, 694 CgHioO.,, 9 (CeHio6oN.),s.io, 773 C6H10O3N4, 731 CgHi-.O^, 97 CgHioOe, 30 C,jH,.0-, 31, 32 CeHigOiN, 692 CgHigOoN, 693 CgHisO-.N, 33 C6H13O9P • 3HoO, 34 CeHi^OoNo, 695 CfiHi^OoN^, 696 C6H14O3N., 697, 737 C6H14O6, 35 CeHi^N, 647 C^H^OXl^, 378 C^H-.O^N, 968 C-H,;O..N..S.„ 913, 1295 C.II.At, 186, 379, 491 C-H,jO,, 186, 380, 381, 492, 867 C,H,;0,„ 382 C;H-O..N, 186, 698, 699, 700, 701 C^H.O.N-,, 1160 C^HsOi, 186, 294, 383 C,HsO-„ 296 C-H.,0,N, 712 C,H,„0,, 297 C-H,„0,;, 298 C-H,, ,.,0.„ 1218 C^HhOuP, 98 C,H,.0-„ 299 C-HioOfi, 300 C7H13N3, 653 C7H13O0N, 702 C7H14O4N0, 703 C7H14O4N0S, 704 C7H14O5N0, 717 C.Hi^O,, 36 C-H17O3N, 654 C-H10N3, 655 CgHga.N, 192 CgHgOoN, 190 C8H5O3N, 191 CgHgOS, 895 CsHgO^, 186 CsHeO,, 186, 372, 384, 385 C.,H«Oe. ^85, 373, 386, 387 CgH^N, 933 CgH.OgN, 896 C8H7O4N5, 1049 CsHsOo, 619 C.HsOoNoSo, 914 C,sH,,03, 186, 388, 389, 390, 493 CsHsO^, J 86, 391, 392, 494, 495 CgHsO,, 497, 1164 CsHgO.N.S, 1091 CSH1063, 295 C,sH,,A, 144 C8HioO,;NP, 969 CsH,iN, 656 CsH„ON, 657 CgHuOaN, 970 Pfizer Handbook of Microbial Metabolites 750 CgHioOsN.S, 897 CsHi,0,N, 1237 C,hHi40, 10, 46 C^Hi40..S.>, 99 CHi^O,, 37 CsHi-,ON-„ 915 CsHieO.N.., 713 C9H4O, 193 CH^Ofi, 374 C9H4O7, 375 Cc,H„0, 194 CgHyO., 620 CgHsOs, 195 C9Hh04, 145 CgH.Os, 393, 394, 412 CoHsO-, 395 CoHcON, 621 CcHcO^N, 971 CgH^O-.N, 396 CgHgO.N-,, 1050 CoH^o6.,N..So, 916 C9H10O,, 622, 853 CoH.oO^, 146 C<,Hj„0„, 147 Cc,HiiO.,N, 705 Cc,H„0,N, 706 C,,HnOHN,-i, 1051 (C.,Hi.O:iN.),„ 1066 Cc»Hio6«N., 1009 Cc,Hi.O;N., 715 CoHijA^N;,, 1010 Cc,Hi30,N, 714 Cc,Hi:A.N,P, 1011 CgHiA-.N^, 898 CoHiAsN.jP, 1012, 1013 CoH^.A.Na, 707 Cc,H,50.N,S, 708 CcjHi.A^NS, 899 Ci,Hi,07N3, 715 C.,H,,Or.N, 716 CgHiA.N, 658 CcH.oOo, 1117 C^H.-O-NCl, 659 C.,H.,,04N,, 737 CioH.iO, 197 CoH.O.s, 198 Ci,H,04, 199 CioHeO,, 516 CioHsO, 200 CioH,0„ 201, 202 Ch,HsO,, 397, 868 CjoHsOfi, 185, 398 CioHh04-CioHi„04, 376 CioH.,ON-„ 1030 Ci„H.,O..N, 934 Ci„H,„d, 203, 204 CioHu.O,, 623 Ck,Hu,0-.N.., 920 CtoHioOg, 205, 399, 400, 624, 869 Ci„Hi„04, 206, 401 C,oH,„0-„ 186, 402 CioH^.Ai, 148, 185, 301, 403 CjoH^At. 186 CioH„O..N, 972 CioH.iOeN, 417 C10H1..ON.., 935 CioH,o03, 404, 405 C,oH,..04, 406, 498, 854 Ci„Hi.,04N.., 1054 Ci„H,o04N4, 1031 CioH,..0,;, 149 Ci„H,..OsN.., 1014 C,oH,,0-N, 407, 973 Ci„H,:AsN-., 1032 CioHi,,04N,, 1033 CioHi^O.-^N.,, 1034 CioHi30,N4P, 1035 CioH,404, 499 CioHi40„ 150 CioHi40,N,P, 1036, 1037, 1038 CioH^40sN,P, 1039 CioHiA^N, 151 CioH,oO,N..S, 900 C,„H,«0„ 417 C,oH,,.04, 100 C,oH,fi04N..S, 901 Ci„H,,O.N„ 717 Ci„Hi,,0i:iN-,P3, 1040 C^oH,,0.iN.HS, 718 CioHiAaN,., 902 Ci(,Ho,)Oc,, 38 Cn,H.,,.N4, 660 C„H,0,sNCl,, 917 CiiHA'., 207, 208, 517 751 Empirical Formula Index CHH^O,. 210. 518 C,..H,,0,N.., 989, 990 C„H,„6.., 211 C,..H.,„0,N... 991 C„H,oO,, 209 C,..H.,,0,..N,P.., 1015 CnH,„0-„ 108 C,..H...,0,„N, 40 C,,H,„0,-,. 109 C,..H.,.,0„, 41,42, 43 C,,H,..0.,N.., 709 C,..H.,..0,.., 44 C,,H,..0;, 212. 025 Ci,H,b..N.., 997 C„Hi..O-„ 410 Ci.,H.,ON.,, 998 C„H,.,0,N..CL, 626 C,,H,„0.„ 218 Cl,H,.,0,;, 411 C,,H,..N..O, 1000 C„Hi.,0,, 412 C,,H,,0,, 1144 CnH,,O.N • H..SO,, 1183 C,.tH,,0,N..S.., 938 CnHi,0,N-„ 1041 C,,H,,0,, 872 C„Hi40„ 152 C,,H,-,0-,N, 1312 CnH,,0,N4, 1299 C,,H,sO,.N„ 1053 C,,H,-,0,. 1292 C,,H.,„.,..0,;N.., 1065 CnH,,0.,N, 1125 C,,H....6,sN,S.., 722 Ci,H,-,0-,N-,. 1042 C,,H.,,0,N.., 1173 C,,H„0,N,;S, 1043 C,,H..,Oi,N,P.„ 1016 Ci,HihO„N..P.., 974 Ci,H..,0-,N, 723 CnH,;0,N. 1301 Ci,H..,.0,N.., 711 CiiH,sO.., 101 Ci4Hio04Cl.., 519 CnHisO^N... 710 Ci4Hu,0-„ 4"l4, 886 CiiHisO.N.,, 719 C,4Hi„0-,N.., 1001 CnH.,„O.N 102 C,4Hi„0,„ 416, 500 CnH.,„0,.N.,. 1138 Ci4Hi„0:, 534, 873 C„H.,..0,N..S, 719 Ci4Hi„0s, 501, 502 C,,H....O,,„ 39 Ci4H„0..N, • 2H..0, 1002 C,..HsbN, 994 Ci4H,..04N4S4, 1295 Ci.H.O.N.., 995 Ci4H,..0-„ 874 Ci.HsO^N.., 996 C,4H,..0,, 628 C,..Hi„0.., 213 C14H1..OH, 186, 875, 1246 C,..H,„0„ 215, 216 C14H1..O,,, 535 Ci..H,„0-„ 413 C14H14O4, 219 Ci..HnO..N.,, 1123 C,4Hi,.0,, 417 Ci..Hi.,0„ 627 Ci4i,H,,0,N3, 1139 C,.,H,..0,, 214 Ci4Ht,0i.>N,, 1285 Ci.,Hi.,0„ 217 Ci4H,sO-„ 1200 Ci.,H,.,0-„ 871 Ci4HisO„N,P, 1044 Ci.,Hi,04N-„ 1298 C14H1CO4N, 1076 Ci-HifiON.., 661, 936 Ci4H.,„0„ 1230 Ci.Hh-.ON.S, 1199 Ci4H.,o04, 302 Ci-.H^jOfi. 153 C14H..0O4N0S, 910 CjoHieO.N^, 1052 C,4H.,oO,N,,, 1121 CioHi.O^N.-P, 937 C,4H..i04N, 1158, 1159 Ci.,HihON4C1..S, 903 CuH.iOoN.S, 905 Ci-.HisO.N^SP.. • HCl, 904 C14H....O4, 303 Ci.,H..uON.„ 986 Ci4H.,..04N..S, 909 CioHooOoN.., 987, 988 Ci4H.,40-„ 1111 Pfizer Handbook of Microbial Metabolites 752 4Ho,06N3S, 724 4Ho50i,N3Po, 1017 4H08O.,, 103 4Ho,03, 104 ^HgO^, 536, 537 .HioOg, 538 ,Hio04, 539 ,Hio05, 540, 541, 542, 551, 856 ^HioOg, 543, 544, 545, 546 ,Hio07, 549 5H10OS, 547 ,HnOoN, 977 -.HioO.N.,, 1141 5H10O4N6, 1055 5H10O5, 415, 550, 887 sHjoOe, 418 ,Hi..O,, 551 5H14N4S..OP,, 1075 5H14O, 3'l9 5H14OP., 419, 520 5H14O,, 521, 1126 -,H,40s, 420, 421, 422 5H15O6N3S, 1003 5H16, 320, 321 gHieOoNo, 304 ,U,,0„ 878 gHieO.NoS.,, 939 5H17O4N, 3"05 gHi^O-.N, 423 ,H,o03No, 1216 5H,o04, 1172 ,Hoi04N, 307 gHoiOgN,, 725 ,H.,..0, 855 ,H.>o03, 855 5H.,.,04, 1177 5H.,304N, 308, 309, 310 5H03O5N, 311, 312, 313, 314 ,H..40.., 46 5H.,40i7N..Po, 1018 gHogOo, 889 .HogOgNo, 1174 gH^gOioN,, 45 -,H3oO.)N.., 1192 eHioOoN.., 940 eHioO,Cl4, 441 eHioOe, 548 6H10O7, 442, 552, 553, 554, 857 eHioO.CL, 424 6H10O5, 555, 556, 858 eHjoOe, 557, 558, 560, 561 gHjoO^, 562, 859 6H14O4, 563, 564 6H14O4N.,, 1231 6H14O.,, 890, 891 fiHi^OgClo, 1153 ,H,40„ 443 6H14OS, 444 6Hi,04N3So, 941 eHi.No, 944 eHisON.,, 945, 946, 947 gHisO,, 220 oHmOoN.., 948, 949 6H18O4N0S, 906 ,H,sO,, 425 gHisO.NoS, 908 eHic03N3S3, 762 cHooN., 950, 951, 952 gHooONo, 953, 954 eH,,o04, 1191 eH_ooO,, 425 gHoiON, 978 eHoiOoN, 979, 980 eHoiOgNgS, 911 6H03O4N, 315 gHogO^N, 314 eHo604NoS, 907 eH.oO^, 105 6H..6O14N0P.,, 1019 gHogOo, 46 6H08O4N4S, 912 6H30O.,, 106 6H30O,, 107 6H30O0, 108 r,H340, 47 ,Hi4No03, 1245 ^HjoOoN., 1300 VH12O7CL, 426 ^HioOgCL, 427 -H10O.J, 5"65 ,Hi403N., 981 7Hi40,Clo, 445 7Hi40«No, 1004 .HieO.No, 997 ,Hi60„No, 1005 7H16O7, 428, 446, 1092 ^HiPeBr, J 86, 431 753 Empirical Formula Index C,,H,-0,.C1. 186, 430 C,„H,40„, 455 C,;H,,0,., 186, 432 C,.,H,,0-,C1,, 456 C,;H.,„0,i. 186, 433 C,.,H„-,0-,Cl.., 457 C,;H..„0,,N^, 1056 C,.,H,,A^ 1308, 1309 C,:H..iO.»N^P, 1057 C,.,H„,0,,, 458 C,;H..-A;N, 316 C,„H,-OsCl, 459 Ci;H..,04, 154 C,.,H,;OsN, 569 Ci:H.,sO,i, 109 C,«Hi,0,, 570 Ci;H..„ON, 768 Ci,Hi,0„ 571 C,,H,„0,, 155 CioHisOs, 460, 461 Ci,H„OsN-,. 729 CioHisO.,, 462 Ci7.isH,i.,,0,sN,„ 727 Ci.,H,sO,o, 463 Ci;H.,o04, 110 Ci<,H,.,0,iN7, 1058 Ci,H,,0-,N,o(S04).., 730 Ci,H..„0„ 464 CisHioO^, 629 C,„H,,A, 572 CmHioO-,, 630 CioHo,0(5N3S..Cl • CCI4, 1277 CmH,oO..N.>, 434 Ci.>H....03, 435 CmHi-O^, 503, 504 Ci<,H....O(,, 322, 323 CisH,..0,,, 505, 892 C,.,H.,.A>N3, 955, 956 CmHioO-, 506 C,.,H.,40-„ 324, 327 C,sH,..0„ 447 CiaH,40«, 325 CisH.oO^o, 448 Cic,H,eO«, 326 CisHu03N,, 1245 CioHoyO.N • HCl, 598 CmHi^O-Cproposed), 522 CigHogOg, 436 Ci.HuO^Clo, 449 C19H30O4, 156, 157, 158 CisHi40s, 450 CieH3404, 159 C1SH14O9, 451 C,,H,,0,, 114 CisHi^Oio, 452 CicHj.O.N, 1252 CisHjA,, 567 C19H3SO,, 115 CisHisOeCl, 453, 566 C<>qHjo04> 523 CigHifiO^, 860 CooH^oO,,, 507 CisHieO^Clo, 1229 C00H13O3N3, 942 CisHisO^, 454 C00H14O,, 599 CisHisOg, 568 C00H14O,, 525 C,,Hoo,06, 317 C20H15O,, 1189 C^sHofA, 317 CooHieO,, 221 CmHogON, 982 CooHieOg, 632, 633 CisHo-.ON, 983 CooHicO,, 559 CisHo-,O..N, 984 CooHieOio, 465 Ci.H.eOaNio, 918 CooHi^O-.Cls, 466 C1SH07O4N, 1303 C.>oH.,oO-, 573 C18H34O0, 111 CooHooOc, 601 C1SH34O3, 112 C.-oH.oO^No, 992 C1SH34O16, 48 C..„H >0-„ 1248 CmH3,0„ 1224 C.,oH.,.,0„ 467 CisHgeOo, 113 C..oH.,..On, 468 CisHaeOnN^, 52 CoH-.A^N^, 1059 CisHggO, 49, 50, 51 CooH..-,ON3, 919 C19H14O5, 631 C2oH,oOeN„ 257 Pfizer Handbook of Microbial Metabolites 754 CooH.sOs, 328 C20H04O6N, 634 C>oH..,Oic,N,oP, 1045 C02H04O8N., 613 CoHogOoN, 985 C00H04O9N.,, 610 C..0H..9O7N • HCl, 596 CooHogO.,, 861 Co^HogOgN, 1180 CooHofiOg, 471 C,„Ho90,sN • HCl, 597 CooHogO-.No, J 86, 993 C00H30O0, 329 CooHoqO-.N^, 1047 C20H30O3, 330 CoHooO^NgS, 1169 C00H30O7N4, 1167 C..o.o4H3o.340,s 9, 1307 C.,oH3..0s, 1236 C00.03H30.34O11, 1132 (aoH3..09No)„, 1222 C...H34O,, 1247 aoH3609Ns, 731 C00H36O6, 1233 C.0H44N0, 662 Co.,H3.s06, 119 CoiHis04, 222 C.-.H^sOeNo, 738 C01H00O3, 223 C.,..H3904N,, 1129 C01H00O7, 574 C....H40O-, 120 C.iHoiOgNoCl, 602 C....H40OSN4S.., 719 CnH^oOg, 880 C0..H44O6, 121 C2iHo<,05Cl, 881 C..3H00O6, 611 C01H00O7, 1176 C.,3H.,40-„ 883 CoiH^oOg, 600 C.,3H..-,0,N • HCl, 612 CoiHooOgNoCl, 603 C^H.-oO,, 884 C01H04O7, 469 C.,,H ,,0,, 474 C21H26O,, 882 C23H.,80,, 472, 473 C21H27O14N7P2, 975 C03H..9O1.N, 57 C.,H„„0,-N.P„ 976 r H n t\t isi. C21H08O17N-P3, 976 CsiH.gOnN, 1297 C2.sH29-3l6,N3, 1314 ^21-29-11-, — / C23H4,Oj4N5, 59 CoiHgoOg, 116 C.,3H4,.OioNp., 60 C2iH3,08N, 1180 C" H O 475 C2.H3eO,N, 1276 C H o" 476 C.oiH360,eN,SP3, 1046 T H O N T 7fi9 C.1H38O6, 117 C24H23O5N5S4, 762 C:;H38o!, 118 C24H2SO4N4, 957 C"oiH390ioN„ 54 C2,H2909Nio, 1304 C01H39O13N,, 55 C24H30O7, 477 C21H41O12N7, 56 - C24H3oO,s, 478 CooHifiOe, 575 C24H3iO-N,Clo, 739 C22H16O7, 604 C..4H33O2N-, 9'22 G22HieOi2, 470 C24H36O9N2S, 923 C-HigOe, 1140 C.4H36 40O9N0S, 258 C22H20O,, 1134 C24H;sOio, 1193 C21.H20O8, 605 C24H40O6, 1232 C2i.H2o09, 606 C24H41O6N, 268 C22H20O10, 576 Co4H4.,06No, 740, 741 C22H2iO,N2Cl, 607 C:.4H4:0,N:, 750 C22H230,N, 870 Co4H4;0-„ 122 C22H230,N2Br, 609 C:,4H5oO;N8, 742 C02H23O8N2CI, 608 C.>4H4oO-N2, 750 755 Empirical Formula Index C.^H^.OhN,, 729 C..4H-,„0.,, 1198 C..-,H,„0., ■ H..O, 508 C..-,H..„0,iN.., 614 C..,H....O„„ 479 C..-,H..-,0,;N, fi-iS C..-,H..sO^, 480 C..-,H,oO,, 437 C.,,Hi„0.. 481 C..-,H,,0,;N,. 743 C..,H,.,0:, 482 C..-,H,..0,. 483 C..-,H,-,ON,. 924 C..-,H,,0,N-„ 1020 C..,H,,,0,N-,C1.>- 751 C..-,H,.,0,N, 259 C..-,H4„0:, 1087 Co-,H4,0„N, 260 C05H43O-N, 261, 262, 263, 1106 C^H^^O.N.., 748 C.>-,H4-,OioN, 264 Co,H,60sNCl, 265 C..-,H,,Oi,N-„ 64 C..-,H-,oO.., 123 C..,,Ho,Oio, 484 C.,,H,,.06No, 224 C.,.H;^„Os, 485 aeH^.Os, 486 C26.l'tH3oO,kN,, 752, 1103 C.,,.H3,0,N;^, 746 CooH3,Oi,,N, 577 C0CH34O7, 318 C,oH„.0<,N., 270, 272 C,6H3,0,;N, 628 CogHasO;, 578 C..,,H,„07, 579 C.iH^.Os, 1262 C,,H,,.,,0,N,, 749 C,eH„;0,N., 747 Co,,H4,0,;R., 1113 C..,;H,,.0.., 124 Co,H3,0i,N,Po, 1060 C.7H3SO4, 160 C,,H3,0,„N,„ 753 C,;H4oO,N,sS3, 1287 C,-H4,A.N.'. 1310 C27H40O1GCI, 1156 C.HnO, 331 C..-H,.,0, 332 C.-H^.O, 332. 333 C..;H,-OhN. 266 C..;H,.,0,-N:, 65 C.-Hv.O.., 125, 126 C..sH.,.iO,,N, 636 C.H.sO,.., 1151 C.,sH,..0„ 1235 C..,H.,.,0.„ 487 C..sH,,jOsN.„ 754 C..sH,-O..N„ 943 C.,sH,,0,„N,;, 1167 C..sH,„O..N.., 269, 1099 C..sH,oO„N,;, 1021 C.,sH,.,0, 334, 335 CsH^AiN, 1122 C.H.^O, 336, 337, 338 CsH^.O,, 339 Co^H4,.0, 340, 341, 342, 343 C..hH4,.03, 344 asH4,Ai, 1234 CosH^hO, 345 C..sH4<,0,N, 274 C.sH^cAsN, 267 C9sHr;g, 11 Co,H..oO,„, 1107 C.^H^Ar-Ng, 1061 CkjH^^O.,, 488 CocH.eO,,, 489 CoH^.N^SOe „ 1102 C.^HasO-NgS, 1089 C0CH40O-, 128 CKtH^.O-NgS^Cr, 1313 Co^H^.OgNfi, 1022 C09H44O9, 1073 CgH^eOo, 1244 C.>9H-,oO.., 438 C3,HisOio, 580 CsoHisOii, 581 CsoH^sOi,, 582 C;{oHihO]s, 583 C3„H..„0,.., 584, 888 C.3oH....O.s, 585 C3„H...,Oio, 586 C30H....O1.,, 587, 588 C3oH.,,Oi4, 1152 CaoHo.Oio, 589 C-30H28-30O11, 592 Pfizer Handbook of Microbial Metabolites 756 C30H34O4N4, 243 C,,H,,0,N4Fe®0He, 925 CsoHggOnN, 615 C^.H^.O^N^, 926 CaoHg.OsN,, 958, 959 C,,H,,.,,0,,N, 1280, 1281, 1282 C3oH3,OiiN, 616 C,,H,,Oj,N, 226 C30H46O0, 346 C3,H,,0s, 1088 C30H46O3, 347 C.,4H,,N0c,, 1162 C30H48O0, 348 C34H,,,OioN3, 921 C30H48O3, 349, 350, 361 C3-,H,c,0,N,, 966, 967 C30H48O,, 225, 1228 C3r,H4(.09NgS, 757 CsoH-^o, 351, 362 C3,-,H,,OioNsS, 756 C3oH,oO, 352, 363 C3,H,oOioNo, 1279 CgoH.oOioN.., 1112 C3,H530i4N (proposed), 227 C30HV.O, 364 C3,H5c06Ns, 786 C30H50O0, 365 C35HeoOi3, 239 CaoH.oOa, 353, 366 (C35HeoOi4No)„, 1217 CgiHo'eOn, 591 C3,H,i0^oN, 276 C3iH;o.3'^Oi4, 1274 C36H3SO8N4, 927, 928 C3iH3e6„N.., 885 C36H4,Oi4N9S, 1241 C3iH390,N-„ 960, 961 CgeH^Oj^N, 228 C3iH3,Oc,N3, 1204 C36H,gOio, 1187 C31H46O4, 354 C3oH,oOioN4, 758 C31H4SO3, 355 CgeHenOigN, 277 C31H48O4, 356 C36H66O10N6, 759 C3iH,o03, 357 C3eH,403, 66 C3iH,o04, 358, 359 C3,H4gOo, 161 C31H60O, 13 C37.46Hci-750a3-16N, 1238 CgiHo-.O, 14 C37H61O14N (proposed), 240 CaiHeoOo, 129 CsTHeo^eOi,, 127 CgoHooOg, 509 C37H67O12N, 278 C32H26-3oC)l4' 526 P H O N 27Q ^S2"-28^U^ ^A" p TT f) j^ 7Ke r" 14 n 11^^ '-'38-39"47-48^9^^6' ' -'-' *--32J^30 32'-'l4. A*^^ P H O IV «! 1987 C30H32O14, 439, 527 C38H55U7^Jn=»4, 1^87 C3IH34O14, 440 C3.,H57.6i07.8N7S, 760 C3oH4iO,N5, 962, 963 C3SH63O14N, 280 C30H40O8, 367 - CgsHo^Oi^N, 281 C32H4e09No, 1279 C39H51O14N, 593 C32H4HOS, 368 C39H58O4, 512 C32H,40c,N, 1070 C39H6,OnN5, 763 C32H54O10, 238 C39H690„, 1225 C32H(joOi4, 130 P H O N 929 '-'32^62'^3' ^-^A r" T4 n lfi9 PHD 1^9 ^40'^4S^4) ^"'^ '--32rt64^3, '-^-^ P T4 n Ifi^ C33H3oO,4N3, 1259 C40H52U0, 163 CooHo.O.N,, 964, 965 C40H,,,, 164, 165, 166, 167, 168 C33HV.,04, 360 C4oH,eO, 169, 170, 171 C33-38H54-660n-i3N, 275 C4„H,,02, 172, 173, 174 C33H60-62O14N, 1143 C4oH,70iiN„ 764 C34H.>60n, 528 C40H60, 175 757 Empirical Formula Index C4oH,..iO,oN,oSFe, 765 C40H,,,, 176, 177 C4oHli40l3' 11^8 C40.412Hot.7iOh>N, 1083 C4oH(;s, 178 C,oHe,OioN„ 767 C^oH.oOsNP, 136 C4iH-,^0.., 532 C4,H-,,0, 179 C^iH-.^O... 180, 181, 182 C41H00O, 183 C4iH6c.ToO,4, 229, 1077 C4oH,.,0„;N, 284 C4..H,;oO.., 184, 185 C^oH^-Oi-.N, 282 C4oHfiT0ie,N, 283 C4oH:30ieN, 1084 C4oH,-,0-,N, 133A C4oHs-,0eN, 133B C43H-1O17N (proposed), 285 C44H-,90a8N±CHo, 617 C44H,oOioNs, 770 C44H6-,0,N<„ 787 C44Ho„04, 513 C44HS9O5N, 134 C45H5sO„Ns, 1205 C4,H,sOi-,N.., 286 C4,H,90i,N, 290 C45Hs50ioN,3, 771 C46..2H8.10N4.T, 1118 C46H30O10, 511 C46H64O0, 531 C46H730o„N (tentative), 248 C46H7-O19N (tentative), 230 C46H80O13, 1086 C4vH,,OioN5, 772 C4,HsoO,fiNo, 287 C48H6oOie, 1166 C4sHs..0ifiN.,, 288 C4c,H6i0..4Ni3, 1062 C4c,H630i8Ni3S, 774 C49H74O4, 514 C49-50H87.9lOi8N, 291 C,oH„oO,oN,,S,, 775 Cn2H,,,0,..N„, 744 C52H,0404, 137 C,.3Ho303,N,„ 1243 C,3Hi,,,0,3Ni„, 776, 777 C54HciO,s,Ni3, 1214 C54H8,04, 515 C54H8.O18N0 (proposed), 250 C55Hv40,;N4Mg, 930 C56H,,0,N,H, 778 CsgHsoO,, 533 CceHyeOiaNi., 779 CseHae-y^OiaNie, 781, 782 (C56-6oH96.iy40.9.3i ),Mg, 1110 C56-63Hu,5-ll-Ooo.22N3, 1196 C57H86O10N10, 811 CggHssOieNi,, 798 C58Hio20,N4, 1142 CggHseOieNjo, 812 C59H87O17N10, 802 C59H88O16N10, 806 CsoHooOifiNi,, 800 CeoHooOieNi,, 799 CeoHgoOiyNio, 788 C60H128O32N00, 790, 1257 C61H89O17N10, 803 CeiHgoOieNio, 794 C61H90O17N10, 805 CeiHgoOieNi,, 801 CeoH^oOieNio, 795 C6,HssOi4Ni4PCo, 931 C64H9„Oi6N:o, 793 C,,H,,0,,N,,, 807 C,,H,,0,,N,., 796 C65H8,03oNi3, 813 CesHasOieN.o, 797 C65-67H96-104O:,Ni8, 1226 CeeHse^isJ^iS' 791 CecHioaOir.Ni.S, 814 Ce8H880:3Ni4, 792 C84Hi7404(±5CHo), 138 Ci4sH,,„0,,N„„ 375 Ci86H366O,7(±10CH,), 139 MICROORGANISM INDEX Boldfaced numbers are entry numbers of metabolites pro- duced by a microorganism. Italic numbers are page numbers and indicate mention in a chapter or section introduction. The appendixes and addendum are not indexed. Absidia ramosa, 934 Acetobacter acetosum, 72 melanogenum, 21, 25 spp., 31, 32 suboxydans, 16, 26, 82 xylinum, 512 Acremonium sp., 501 Actinomyces atroolivaceus var. mutomycini, 1218 fiavochromogenes var. helio- mycini, 1171 (Streptomyces) ftavus, 12 genus, 118 globisporus, 1097 longispori, 1187 Actinomycetaceae buchanan, 713 family, 118 order, 118, 119 Actinomycete, 14, 334, 1252 Actinoplanaceae family, 118 Actinoplanes genus, 118 Aerobacter aerogenes, 19, 143, 306, 317, 449, 528, 532 Agaricus campestris, 707 (Clitocybe) nebularis Batsch., 1006, 1007, 1023, 1031, 1033 Agrobacterium tumefaciens, 51, 111, 114 Agrocybe dura, 190 Alectoria sp., 860 implexa (HofFm.) Nyl. f. fuscid- ula Arn., 452 japonica Tuck., 487 ochroleuca (Ehrh.) Nyl., 363 ochroleuca Mass., 467 sarmentosa Ach., 487 zopfi Asahina, 450 Aleuria aurantia, 164, 165 Algae, 24, 35, 43, 212, 213, 231, 301 Allomyces sp., 1276 arbiiscula, 166 javanicus, 165, 166, 168 macrocygna, 166 moniliformis, 166 Alternaria radicina, 871 solani Ell. and Mart., Jones and Grout, 116, 1074 tenuis Auct., 151, 415, 416, 418, 419, 420, 421, 422 Amanita mappa, 661 muscaria (Linn.) Fries, 43, 74, 508, 537, 641, 650, 658, 659, 707, 1023, 1024 phalloides, 11, 47, 343-345, 351, 652, 756,757, 1198 Amphierna rubra, 29 Anaptychia hypoleuca, 365 speciosa, 365 Anthomyces renkaufi, 29 Anthurus aserioformis, 168 muellerianus, 638 Anzia gracilis, 477 leucobatoides f. hypomelaena, 477 opuntiella Miill. Arg., 477 Armillaria mellea, 20, 537 Arthrobacter sp., 1179 Ascomycetes, 1056 Ashbya gossypi, 557, 558, 721, 1056 Aspergilli, 19,418, 1049 (white), 81 Aspergillus amstelodami (Man- gin) Thom and Church, 546, 552 candidus, 872, 1125 chevalieri, 555 citricus (Wehmer) Mosseray, 516 759 Microorganism Index Aspergillus amstelodami clavatiis, 105, 852. 8(i7 elegans. 1107 flavipes (Bainier and Sartory) Thorn and Church, :^91, 429 flavus, 24, 73. 865, 910. 986-989 fimiigatus Fres., 79, 318, 335, 336, 339, 496, 497, 938 fumigatus mut. Jielvola Yuill, 367 giganteus, 867 glaucus, 435, 436, 555, 560, 563, 564, 852, 865 itaconiciis, 1181 mangini, 435 vielleiis Yugawa, 399 nidulans, 50, 456, 457, 466, 547 niger, 29, 31, 42, 68, 82, 84, 88, 92, 95, 143, 334, 852, 890, 901, 934, 302 niveus, 872 ochraceiis, 144, 399 onjzae, 73, 312, 537, 683, 694, 702, 852, 865, 927, 1025, 1237 parasiticus, 24 quadrilineatus Thorn and Raper, 401, 547 ruber (Man gin) Raper and Thorn, 555 sclerotiorum, 990 spp., 35, 78, 86, 310, 404, 435, 436, 778 sydoivi, 135, 686 tamarii, 865 teireus mutant, 89 terreus Thorn, 20. 80, 83, 295, 394, 424, 426, 427, 492, 867, 872, 1293 ustus, 412 versicolor, (Vuillemin) Tira- boschi, 543, 892 wentii, 17, 852 Auxotrophs, 143 Azotobacter vinelandii, 237, 448, 514 Bacillus aerosporin, 780 alvei, 830 anthracis, 703 Bacillus aerosporus brevis, 531, 786. 787. 789. 791. 792, 826,827, 1157 brevis var. Gause-Brazhnikova, 788 bruntzii, 436 cepae, 1090 cereus var. mycoides, 971 cereus var. terminalis, 968 circulans, 776 circulans mucoid variant, 779 coli, 17, 18 colistinus, 825 krzemieniewski, 779 laterosporus, 1182 licheniformis, 814, 844-846 megatherium, 37, 440, 714, 968 mesentericus, 19, 238, 763 polymyxa, 19, 785 prodigiosum, 919 pumilis, 762, 1091, 1255 pyocyaneus, 1000, 492 sphaericus, 479, 968 spp., 306 subtilis, 17, 19, 185, 396, 422, 483, 814-816, 836-839, 842 1078, 1115-1117, 1151, 1157, 1269 subtilis var. aterrimus, 1109 Bacteria, 14, 15, 17, 35, 87, 154, 185, 237, 290, 291, 308, 316, 332, 333, 436, 480. 488, 492, 496, 508, 509. 531, 532, 969, 1060, 1062, 1040 Baeomyces fungoides Ach., 461 roseus Pers., 461 Basidiomycete, 1085 B-841, 220 Basidiomycetes, J 07, 238, 291, 427 Blastomyces brasiliensis, 3 dermatitidis, 3 Boletus appendiculatus, 638, 652 badius Fr., 537 chrijsenterou Bull., 537 edulis Bull., 2, 638, 641, 643, 650, 652, 656, 707, 934, 1023, 1026, 1027 elegans, 650 Pfizer Handbook of Microbial Metabolites 760 Boletus appendiculatus luridus Schaeff. ex Fries, 537, 652 luteiis, 650, 656 qiieletii, 652 regius, 652 sanguineus, 652 satanas Lenz, 537 spp., 78, 305 subtomentosus Linn, 537 versipellis, 641 Botrytis alii, 302 cinerea, 1 Buellia canescens (Dicks.) De Not, 441 Caldariomyces fumago, 293 Calicium chlorinum Korper, 631 hyperellum. Ach., 636 Calocera viscosa, 345 Calonectria sp., 997 Caloplaca elegans (Link), 555 Candida flareri, 558 guillermondi, 558 parapsilopsis, 558 pulcherrima (Lindner) Win- disch, 991 Cantharelhis cibarius, 164-167 cinnabarinus , 163, 165 multiplex Underw., 507 spp., 168 Carpenteles brefeldianum Dodge (Shear), 430 Cephalosporium sahnosynnema- tum, 312, 367-371, 905, 911 spp., 368 Ceratostomella fimbriata, 620, 851, 853-855 Cetraria collata Miill. Arg., 488 crispa Nyl. ( = C. tenuifolia Howe), 158 crispa (Ach.) Nyl., 363 cucullata (Bell.) Ach., 363 delisei (Bory) Th. Fr., 363 hiascens, Th. Fr., 363, 476 islandica (L.) Ach., 157, 363, 470 islandica Ach. var. orientalis Asahina, 158 Cetraria collata Miill. Arg. islandica F. tenuifolia, 156 japonica, Zahlbr., 489 juniperina Fr. var. tubulosa Schaer, 631 juniperina L. (Ach.), 632 nivalis (L.) Ach., 363, 364 pinastri (Scop.), 631, 632 pseudocomplicata Asahina, 487 sanguinea, 477 sp., 860 tubulosa (Schreb.), 632 Chaetoniium affine Corda, 539, 542, 592 aureum Chivers, 501 cochlioides, 392, 941 indicum Corda, 284, 628 Chlorobacteria, 930 Chlorobiiim spp., 166 Chlorophyll-containing bacteria, 438 Chlorosplenium aeruginosum (Oeder ex Fries) De Not, 528 Chromatium species, 172, 179- 181, 184, 237, 238 Chroniobacterium iodinum, 996 violaceiim, 942 Circinella species, 78 Citric acid-forming fungus, 502 Citromyces spp., 68 strains, 873 Cladonia alpestris L. Rabh, 363 amaurocraea, (Fl.) Schaer., 464 bacillaris Nyl., 464 bellidiflora var. coccocephala Ach., 462 coccifera (L.), 464 deformis Hoffm., 4, 362, 365 digitata, 458 evansii. Abb., 473, 482 floerkeana Sommerf., 464 impexa Harm., 22, 361, 463, 473, 482 macilenta (Hoff.) Flk., 464 mitis Sandst., 117 nemoxyna (Ach.) Nyl., 478 papillaria (Ehrh.) Hoffm., 157 pityrea Flk. f. phyllophora Mudd, 478 761 Microorganism Index Cladonia alpestris L. Rabh polydactijla Flk., 158 pseiidoevansi Asahina, 473, 482 pseudostellata Asahina, 463 rangiferina (L.) Web.. 470 ravqifonnis HofFm., 117 species, 212, 458, 565, 860, 861 squaj7iosa HofFm., 462 strepsilis, 856 sylvatica L. Harm., 361, 470, 664 uncialis (L.) Web., 462 Clasterosporiiim spp., 17, 81 Clathrus ruber, 640, 643 Claviceps purpurea, 43, 48, 291, 336, 341, 344, 351, 465, 47J, 535, 553, 639, 643-645, 647, 648, 651, 652, 654, 656, 657, 668, 673, 693, 944-967, 1133, 1151, 1152, 1274 Clitocybe Candida, 1135 diatreta, 191, 192, 198 illudens, 1177 Clostridia, 237, 449 Clostridium acetobutylicum, 18 butylicum, 30 propionicum, 74 propylbiitylicum, 18 saccharobutylicum, 18 tetanomorphum, 445, 446 Coccifera bellidifiora, 365 pleurota, 365 Coleosporium senecianis, 164-166, 168, 170 Collybia dryophila, 672 Coprinus comatis Gray, 651, 652, 657, 708, 1026, 1027, 1199 miraceus, 672 quadnfidus, 193-195, 201 similis B. and Br., 493 Cordyceps militaris (Linn.) Link, 1032 sinensis (Berkeley) Saccardo, 300 Coriolus sanguineus Ft., 1001 Cornicularia diverqens Ach., 486 pseudosatoana Asahina, 486 Corocyneamembranacea (Dicks.), 859 Corticeum croceum Bres., 219 saliciuu 771 Fries, 223 sasakii, 390 sulfureum (Ft.), 219 Cortinarius cinnaharinjis Fries, 562 cin7iamomea, 638 sanquineus (Wulf.) Fries, 542, 562 Corynebacteria, 51, 54, 121, 437 Corynebacterium diphtheriae, 13, 16, 51, 106, 131, 132, 137, 168 314, 343, 518, 674, 698, 703, 873, 928 insidiosum (McCulloch) Jen- sen, 1179 michiqanense, 163, 168 michiganense mutants, 165 ovis, 132 sp., 548, 1061 Cryptococcus laurentii, 142, 165, 166 luteolus, 165, 166 Cunninghamella species, 78 Curvularia lunata, 1200 sp., 425 Cyanococcus chromospirans, 1000 Cyphelium chrysocephalum Ach., 631 Dacromyces stillatus, 164-166, 171, 173, 176 Daedalea juniperina Murr., 619, 895 Daldinea concentrica (Bolt) Ces. and De Not, 388, 404, 523, 627, 869 Debaryomyces hansenii, 142 Dermatocarpon miniatuTTi (L.) Mann, 36 Dermocybe (Cortinarius) Cinna- momea, 638 Dimelaena oreina, 365 Diploschistes bryophilus (Ehrh.), 444 scruposiis (L.), 444 Discomycetous inoperculate fun- gus, 525, 526 Pfizer Handbook of Microbial Metabolites 762 Drosophila semivestita, 207 suhatrata (Batsch. ex Fr.) Quel., 378 Endoconidiophora coerulescens Miinch, 9, 10 virescens Davidson, 10 Endothia fiuens Shear and Stevens, 580, 586 parasitica (Murr.) Anderson and Anderson, 580, 586, 1144 Enterococcus stei, 556 Erevtothecinvi ashbyii, 516, 529, 557, 558, 560, 1008, 1052, 1053, 1056 Erwinia chrysanthemi, 1179 Escherichia coli, 98, 104, 143, 237, 238, 290, 296-298, 301, 306, 307, 310, 312, 314, 341-343, 423, 483, 509, 527, 531, 532, 537, 554, 556, 557, 662, 674, 691, 703, 898, 933 Aerohacter aerogenes type of bacterium, 841 mutant, 99, 344, 444, 460 Evernia divaricata L., 469 mesomorpha f . esorediosa Miill. Arg., 469 prunastri L., 406, 446, 459 sp., 860 vulpina L., 631 Fistulina hepatica, 22 Flavohacterium marinotypicum, 186 sulfureum, 186 Fomes fomentariiis, 338 juniperimis (Polyporus), 1164 laricis, 120 • officinalis, 120 Fremella mesenterica, 165 Fungi, 15, 35, 68, 81, 83, 90, 95, i54, 185, 212, 213, 231, 232, 291, 299, 303, 532, 564, 683, 702, 1058 Fusaria species, 78, 335, 336, 479, 738, 741 Fusariiim bostrycoides Wr. and Rkg., 522 bulbigemim, 1244 Fusarium bostrycoides Wr. and Rkg. bulbigenum Cke. et Mass. var. lycopersici (Bruchi) Wr. et Rkg., 973 culmoriim (W.G. Sm.) Sacc, 584, 887 gravtinearum Schwabe, 887 heterosporum Nees, 973 javanicum Koorders, 520 lateritiiim, 740 lycopersici, 301, 715, 1189 moniliforme, Vl% orthoceras App. et Wr., 973 orthoceras var. enniatinum, 740 oxysporum, 80 scirpi Samt. et Fautr., 740 solani (Mart.) App. and Wr., 521 sporotrichiella var. poae, 1251 vasinf ectum Atk., 973, 1189 Ganoderma oregonense, 1236 Geaster fimbriatus Fr., 345 Gibberella baccata, 1113 fujikuroi (Saw) Wollenweber, 321, 324-326, 534, 852, 972, 973 saubinetti, 82, 887 Gliocladium fivibriatiim, 938 roseiim Rainier, 498 sp., 236 Gluconoacetobacter liquefaciens, 72, 405, 862-864 roseum, 407, 865, 866 spp., 404 Gram-negative bacteria, 342 -positive bacteria, 119, 342, 344 Grifola confluens, 46 Gyranoasciis spp., 867 Gyvinosporavgiiini juniperi-virgi- vianae. 164-166 Gyrophora deiista (L. ), 479 esculenta Miyoshi, 475 polyphylla (L.), 479 proboscidea L., 475 vellea (L.), 479 Haematomma coccineum, 365, 857 leiphaermim, 365 763 Microorganism Index Haematomma coccineum porphiiriutu (Pers.), 365, 857 sp., 8(i() ventosinu, 121 Hauscnula auomala, ()99 suhpclliculosa, 142 Helicohasidium monpa, 83 HelmintJiosporium avenae Ito and Kurib.. 544 catenarium Drechsler, 541, 546 cynodontis Marignoni, 541, 544 eiiclaevae Zimmermann, 544 grammeum Rabenhorst, 541, 546 leersii Atkinson, 579 ravenelii, 886 triticividgaris Nisikado, 541, 546, 549 velutiniim Link, 546 victoriae, 544, 768 Histoplasma capsulatum, 3 Hijdnurn aspratum Berk., 2 aiirantiaciim Batsch., 509, 511 imbricatum L., 345 spp., 507 Hydrogenornonas species, 238 Hypholoma capnoides, 537 Hypochniis sasakii Shirai, 390 Inocybe patoullardii Bres., 1138 Kloeckera brevis, 142 Lactariiis deliciosus, 319-321, 638 helviis, 638, 710 rufus Scopol., 112 spp., 9, 305 turpis, 537 vellereus, 537, 638, 641 Lactobacilli, 51, 75, 87, 119, 154, 449, 513, 561 Lactobacillus acidophilus, 1019 arabivosus. 111, 114, 514, 556, 931, 1015, 1017, 1038 casei. 111, 114, 333 helveticiis, 334 leichmannii, 445, 516, 552 pastorianus var. quinicus, 299 Lecanora atra (Hudson) Ach., 488 cpatwra Ach., .365, 635 gangaleoides Nyl., 22, 449 gru7uosa (Pers.) Rohl, 488 parella Ach., 442 sordida, 365 sp., 110, 860 sulfurea, 365 thiodes, 365 Lentiniis dactyloidcs, Cleland, 357 degener Kalchbr., 493 lepideus Fr., 619, 622-625 Lenzites spiaria (Wulf), 1185 thermophila, 568 Lepiota clypeolaria, 638, 641 Lepraria candelaris Schaer., 630 citrina, 633 fiava (Schreber. ) f. quercina, 632 latebrarum, 365 Leuconostoc mesenteroides, 41 Lichens, 35, 68, 80, 121, 154, 157, 190, 212. 213, 231, .336, 400- 402, 4,92, 496, 1059 Lobaria oregana Mijll. Arg., 455 pulmovaria (L.) Hoffm., 22, 447, 455, 507 pulmonaria, HofTm. f. tenuior Hue., 484 pidmonaria, var. meridionalis (Wain.) Zahlbr., 475 retigera Trev., 507 Lycoperdon gemmatum, 652 piriforme, 652 pratense, 537 "M-14" strains, 1142 Macrosporium porri. Elliott, 556 Marasmivs comgenus, 1191 gramineiim Lib., 517 peronatus, 652, 656 ramealis, 200, 397 Merulius lacrymans, 210, 212, 215 Metarrhiziiim glutinosum, 1166 Micrococcus lysodeikticus, 174, 305, 537 sp., 306, 761 Pfizer Handbook of Microbial Metabolites 764 Micrococcus lysodeikticus tetragenus (pink type), 168, 170 varians, 680 Micromonospora genus, 118 globosa, 334 sp., 1203 Microsporum canis, 1063 gypseum, 1063 Mitrida paludosa, 165 Molds, 14, 43, 50, 154, 308, 313, 315, 316, 436, 492, 496, 969, 970, 975, 976, 1018, 1040, 1060 Mollisia caesia, Sacc. sensu Sy- dow, 519 gallens Karst., 519 Monascus purpureus, Wentil, 882, 884 rubTiginosus Sato, 882 rubropunctatus Sato, 880, 882 Monilia formosa, 79 sitophila, 165 Monosporium bonorden, 1092 Mucor species, 78 hiemalis, 91 mucedo, 303 ramannianus , 1263 stolonifer, 80 Mushrooms, 43 Mutinus caninus, 638, 652 Mycobacteria, 43, 51, 55, 121, 188, 237, 531 Mycobacteriaceae family, 118 Mycobacterium avium, 50, 556— 558 battaglini, 189 genus, 118 laticola, 162, 1049 marianum, 189 phlei, 50, 160, 164-166, 168, 171, 173, 174, 176, 177, 178, 185, 187, 188, 530, 697, 772 sm.egmatis, 238, 558, 931 tuberculosis var. hominis, 34, 50, 51, 66, 115, 122-126, 129, 138, 139, 238, 342, 407, 518, 533, 703, 708, 928, 1050 Mycococcus genus, 118 Mycotorula lipolytica, 699 Myoporum spp., 855 Nectria cinnabarina (Tode) Fr., 973 Nematoloma fasciculare, 656, 657 Nematospora coryli, 142 Nephroma antarcticum, 365 arcticum, 365 laevigatum, 365 parile, 365 Nephromium lusitanicum, 561 Nephromopsis cilialis Hue., 487 endocrocea Asahina (= Cetraria endocrocea (Asahina) Sato), 154, 155, 552 stracheyi f. ectocarpisma Hue., 118, 156, 159 Neurospora crassa and inutants, 91, 97, 143, 164-168, 175-178, 184, 187, 238, 291, 301, 305, 310-312, 482, 512, 525, 532, 641, 646, 649, 655, 660, 676, 687, 690, 1014 crassa mutants, 164, 176, 311, 312 sitophila, 178 Nocardia acidophilus, 218 formica, 730, 915 gardneri, 266-268 genus, 118 lurida, 1269 mesenterica, 893, 1098, 1197 narasinoensis, 1227 rugosa, 528 sp., 893, 1226, 1278 Ochrolechia pallescens, 475 Ochromonas malhamensis, 1051 Oidiodendron fuscum Robak, 878 Oospora colorans van Beyma, 501 sulfurea-ochracea, 428 Ophiobahis miyabeanus, 1235 Oxidative bacteria, 44 Pachybasium candidum (Sacc.) Peyronel, 538, 539 765 Microorganism Index Pachiima hoelen Rumph., 358 Paecilomyces, 726 Paecilorntices variotis Bainier var. antiJ)ioticus, 1303 victoriae V. Szilvinyi, 393, 395, 412 Ponaeolus cmyipamilatus, 935 Pannaria fidvescens Nyl., 453 lanuginosa Korb., 453 lanuginosa Ach., 859 lurida Nyl.. 453 Parmelia abyssinica Kremp., 448 acetabulum Duby., 447 borreri Turm., 443 caperata (L.), 118, 451 cetrata Ach., 448 conspersa Ach., 448 formosana Zahlhr., 891 furfuracea Ach., 459, 485 glomellifera Nyl., 481 hypotrypella, Asahina, 465 latissima Fee, 22, 443 leucotyliza, 365, 366 marmariza, Nyl., 448 olivetorum Nyl., 486 perlata Ach., 473, 482 physodes Ach., 459, 465, 485 saxatilis Ach., 448 scortea Ach., 443 sinodensis Asahina, 157 sp., 213, 860 tinctorum Despr., 443 Parmeliopsis spp., 458 Patella vulgata, 683 Paxillus atromentosus (Batsch.) Fr., 505 Pellicidaria sasakii, 390 Peltigera horizontalis, 365 malacea, 365 propagulifera, 365 Penicilliopsis clavariaeformis Solms-Laubach, 585 Penicimum, 28, 497 Penicillium spp., 17, 35, 78, 133, 336, 528, 778, 850 albidum Sopp., 430, 1068 aurantio-virens Biourge, 80, 182, 373, 375 Penicillium spp. baannmse, 144 brefeldianum, 875 atrovcnetuni G. Smith, 570 brevi-compactinn Dierckx, 16, 20, J 85, 386, 398, 402, 403, 433 canescens, 1126 charlesii G. Smith, 140, 146- 149 chrysogenum, 122, 302, S12, 345, 422, 426, 537, 686, 724, 897, 902, 910, 1035, 1037, 1038 chrysogenum. (myceUum), 5 JO, 1044 chrzaszszi, 872 cinerascens Biourge, 145, 497, 938 citreo-roseum Dierckx, 545 citreo-sulfuratum, 872 citrinum, 872 claviforme, 867 crateriforme Gilman and Ab- bott, 109 cyclopium Westling, 20, 81, 144, 302, 303, 409, 411, 536, 545, 977, 981 cyclopiwm-viridicatum, 373, 375 digitatum, 3, 6 divergens Bainier and Sartory, 381, 383 equinum, 867 expansum, 867, 872 fellutanum, 140 flexuosum, 389, 875 frequentans Westling, 302, 873 funicidosum Thorn, 71, 540, 1170 glabrum, 873 gladioli McCull. and Thom, 400, 408, 410 glaucum, 317 griseofidvinn Dierckx, 186, 302, 381, 389, 392, 4J0, 430, 432, 867, 875, 993 herquei Bainier and Sartory, 555, 572, 573 implicatum Biourge, 872, 881 Pfizer Handbook of Microbial Metabolites 766 PenicilUum spp. islandiciim Sopp, 71, 385, 539, 540, 546, 550, 551, 580, 583, 587, 588, 739, 751 islandiciim N.R.R.L., 581, 582 janczexvski Zal., 430, 432 jenseni, 381, 938 johannioli Zaleski, 373, 375 leucopus, 867 lilacinum, 50, 106 lividum, 872 melinii, 430, 867 minioluteum Dierckx, 105, 109 multicolor G. M. P., 881 nalgiovensis Laxa, 566, 567 nigricans (Thorn and Bainier), 430 notatum Westling, 49, 90, 100, 337, 417, 423, 434, 850, 910, 934 novae-zeelandiae , 867 oxaliciim, 68 palitans Westling, 303 patuhim Bainier, 186, 377, 379, 389, 430, 491, 867 paxilli var. echinulatmn, 1153, 1229 pfefferianum, 873 phaeojanthinellum, 872 phoeniceum van Beyma, 500 puberuhnn, 144, 373, 375, 1298 pulvillorum Turfitt, 1254 purpurogenum Stoll, 317, 874 pnrpurogenum Stoll var. rubri- sclerotiiim Thorn, 32, 93 racihorskii Zal., 430' raistrickii, 295, 430 resticulosiim, 712 . roquefortii, 303 roseopurpureum Dierckx, 558 roseo-purpiirogenum, 873 rubrum O. Stoll, 500 rugulosum Thorn, 580, 586 sclerotiorum van Beyma, 164- 166, 881, 883 soppii, 50 spiciilisporiim Lehman, 109 spinulosum Thom, 50, 497, 1093 PenicilUum spp. terlikoiuski Zaleski, 938, 939 stipitatum Thom, 182, 372, 374, 376 stoloniferum Thom, 398, 402, 403 tardiim, 580, 586, 1292 terrestre Jensen, 152 thomii, 144 urticae Bain., 389, 430, 867 viniferum, 79 viridicatuni Westling, 153, 977 wortmanni, Klocker, 580, 586, 1311 Peniophora filamentosa (B. and C.) Burt, 504 Pertusaria amara (Ach.) Nyl., 437 Pertusaria spp., 458 Phallus impudicus, 640, 643, 652 Phlegmacium mellioleus, 652, 656 Pholiota mutahilis, 638, 641, 656 Phoma terrestris Hansen, 569 Photosynthetic bacteria, 437, 438, 926, 927 Phycomijces blakesleeanus, 91, 122, 124, 164-166, 168, 176, 178, 380, 382 Physarum polycephaluin, 353 Physica caesia, 365 endococcina. 365, 595 Phytomonas spp., 25 Pilobolus bleinii, 165 Piriciilaria oryzae, 86, 1245 Placodium saxicolum, 365 species, 213, 555 Pleurotus griseus, 1248 mutilus, 1247 ulmarius, 197, 210 Polyporus anthrocophilus Cooke, 199, 202, 203, 205, 206, 211, 212, 216, 217, 221, 222, 357, 360 australiensis Wakefield, 356, 358 benzoinus, 354 betulinus Fr., 119, 354, 356, 358, 359 767 Microoigai>ism Index Poliiporus anthrocophilits hi for mis. 19(i. 1117 ciunabariuus, 1001 cocci ne us Fr.. 1001 confliu'us Fr., 46, .'{45 eucahiptorum Fr., 357 fumosus Pers. Fries, 494 guttalatus, 204 hispidiis (Bull.) Fr., 357 iuuiperirms, 1164 leucomelas Pers. ex Fr., 506, 510 nidiilans Fries, 504 officinalis ( = Fomes officinalis, Fomes laricis.), 120, 355, 357 pinicola Fr., 346-348, 354 puniceus Kalch., 1001 rutilans (Pers.) Fries, 504 sanguineus L., 1001 squamosus, 537 sulfureus, 291, 345, 357, 537, 638-641, 644, 652, 656, 677, 700, 701, 707, 1023, 1026 tumulosus Cooke, 356, 358, 384, 387, 391 Polijstictus cinnaharinus (Jacq.), 1001 sanguineus L., 1001 semisanguineus Lloyd, 1001 versicolor (L.) Fr., 507 Polystigma rubrum, 169 Poria cocos (Schw.) Wolf, 356- 358 corticola, 208, 209, 213, 214 tenuis, 208, 209, 213, 214 Proactinomyces cyaneus var. an- tibioticus n. sp., 1186 Propionibacteria, 74, 87, 447, 931 Propionibacterium shermanii, 440, 932 Proteus immunitatis anticarcino- matosa n. sp., 1301 vulgaris, 30 Psalliota xanthoderma, 1253 Pseudomonas aeruginosa (Bacil- lus pyocyaneus), 130, J 85, 448, 492, 501, 917, 979, 980, 982-985, 994, 1000, 1002 Pseudomonas aeruginosa aeruginosa strain T-359, 979 aestumarina, 171. 173 antiuiycetica, 824 aureofaciens Kluyver, 997 beijerinchii Hof, 490 chlororaphis, 998, 999 cocovenenans, 128, 1029 fiuorescens, 30, 85 forrnicans n. sp., 67 hydrophila, 19 indigo f era, 1179 pyocyanea, 107 saccharophila , 70 spp., 25, 27, 44, 306, 501 tabaci, 717 viscosa, 759 xanthochrus, 171, 173 Psilocybe aztecorum Heim, 937 caerulescens Murr. var. ma- zecatorum Heim, 937 mexicana Heim, 937 sempervirens Heim et Cail- leux, 937 sp., 936 zapotecorum Heim, 937 Psoroma crassum. Korber, 450 Puccinia coronifera, 164-166 graminis Pers. var. tritici Eri- kas. and Henn., 7 Ramalina boninensis Asahina, 483 calicaris Rohl., 471, 474 farinacea, 451 geniculata Hook et Tayl., 22, 471, 474 intermediella Wain., 471, 474 pollinaria Wests., 446, 454 scopulorum (Retz.) Nyl., 22 sinensis, 22 sp., 860 spp., 454 tayloriana, 22 usneoides Mont., 474 Rhizocarpon geographicum L., 464, 636 viridiatrum Flk., 636 Pfizer Handbook of Microbial Metabolites 768 Rhizopus nigricans, 497, 934, 1055 saponicus, 345 Rhizopus sp., 75, 78 suinus, 934 Rhodopseudomonas spheroides, 182, 183, 438, 926, 928-930 spheroides mutant, 177 Rho do spirillum fulvum, 930 rubrum, 172, 177, 179-181, 184, 238, 930 Rhodotorula glutinis, 165, 168 glutinis var. lusitanica, 101, 102 rwfcra, 161, 164, 165, 168, 175, 185 sanniei, 161, 165, 168 sp., 50 Rhodovibrio sp., 930 Roccella fuciformis Ach., 992 fuciformis DC, 468 montagnei Bel., 20, 110, 171, 468 tinctoria (L.), 110 Russula alutacea, 640 aurata, 640 cyanoxantha, 640 foetens, 652 grisia, 640 lepida, 640 maculata, 641, 652 olivacea, 640 sardonia, 640 spp., 638, 643 turci, 640, 641, 652 _ vesca, 640 Saccharomyces anamensis, 927 carlsbergensis, 336 cerevisiae, 69, 237, 238, 512, 722, 927 fragilis, 238 Salmonella typhi, 306 Sarcina aurantiaca, 165, 168 [utea, 174, 186 species, 90 Schizophyllum commune mu- tant, 940 Scleroderma vulgare, 638 Scopulariopsis brevicaulis, 302 Serratia species, 723 marcescens, 19, 30, 143, 435, 919 marinorubrum, 919 Solorina crocea (L.) Ach., 574 Sparassis ramosa, 406 Sphaerophorus coralloides Pars., 472 fragilis Pers., 472 melanocarpus , 472 Sporidesmium bakeri Syd., 1277 Sporobolomyces roseus, 165 salm.onicolor, 142, 165 Staphylococcus, 343 Staphylococcus aureus, 167, 168, 170, 173, 174, 304, 343, 642 citreus, 186 Stemphylium radicinum Sterad, 413, 871 Stereocaulon exutum Nyl., 480 nabewariense Zahlb., 455 paschale (L.) Fr., 363, 480 Stereum hirsutum, 1172 Sticta aurata Ach., 629, 630 colensoi Bab., 504 coronata Muell., 504 crocata Ach., 630 spp., 643 fuliginosa, 638 sylvatica, 638, 640 Streptobacterium plantarum, 537, 654 Streptococci, 5J Streptococci (Group A), 5J2 Streptococcus albireticuli, 1150 cremoris, 816-819 faecalis, 17, 304, 556 lactis, 816-819 spp., 106, 111 Streptomyces abikoensis, 1094 achromogenes, 1285 acidomyceticus, 1072 afghanensis, 1291 akitaensis, 1067 albo-niger, 1047, 1139 alboreticuli, 242, 283, 284 albulus, 304, 305, 309, 316 a/bus, 1070, 1081, 1139, 1178 769 Microorganism Index Streptornyces abikoensis «//;// ,s-rcscmbling, 1315 fl//;//.s-similar, 1-U5 alhus var. fu7igus, 1069 amhofaciens. 292, 918 atiti})ioticus (Waksman et Woodruff) Waksman and Henrici, 27(). 334, 335, 617, 79'?. 791. 931 arable us, 1140 aureofacietis Duggar, 306, 603, 604, 607-609 aureofaciens strain W-5, 1139 flz/rez^s-resembling, 1242 aureus Waksman and Curtis, 237, 1066 bikitiiensis, 54 bobiliae. 1222 bottropensis, 760 cacaoi, 1129 caelestis, 258 caeruleus. 1123 caespitosus, 1214 calvus n. sp., 1043 canesciis, 256 canus, 833, 1079 carcinomycicus , 847 catenulensis, 61 celestis n. sp., 923 cellulofiavus n. sp., 916 cellulosae, 239 chartreiisis , 439 chattanoogensis, 236 chibaensis, 5 chromogenes, 918 chrysomallus, 334, 764, 793- 795, 802, 803, 805-807, 811, 812 cinnamonensis, 899 cinnamoneus f. azacoluta, 820, 821 coelicolor (Miieller, Waksman and Henrici), 526, 1136 colliniis, 1137, 1271 crystallinus, 1297 diastatochromogenes, 1286 diastatochromogenes-Tesem- bling, 1234 Streptornyces abikoensis echinatus n. sp., 775 endus, 246 erythreus, 277-279 enithrochromogenes, 294 ETH 1796, 306 eurocidicus, 284, 285 eurytherinus, 291 exfoliatus, 1156 fasciculatus, 1022 feZZezzs, 263 fervens, 1160 filipinensis, 238 fiaveolus, 334, 577 flavochromogenes, 259, 1102 fiavofungini, 1143 flavovirens, 334 fiavus, 244, 334, 335 /Zaz^MS, 0-2, 1147 fiavus-parvus, 335 floridae, 727 fradiae, 60, 243, 290, 306, 335, 802, 803, 805-810 fulvissimus, 758 fungicidicus, 1095 fuscus, 1165 galiloeus Ettlinger et aZ., 617 garyphalus, 894 genus, iJ8 glaucus, 923 gr amino faciens, 746 griseocarneus, 55 griseochromogenes, 1120, 1I2I griseofiavus, 1167, 1168 griseolavendus, 736 griseolus, 265, 1076, 1106, 1158 griseoluteus, 1004, 1005 griseoplanus, 725 griseoviridus, 1169 griseus (Krainsky) Waksman et Henrici, 54, 65, 238, 253- 255, 307, 308, 311-314, 407, 440, 529, 765, 766, 885, 91,3, 931, 1096, 1132, 1139, 1159, 1169 griseus var. spiralis, 834 grisews-like strains, 843 hachijoensis n. sp., 251 halstedii, 282, 283 Pfizer Handbook of Microbial Metabolites 770 Streptomyces abikoensis haivaiiensis, 840, 1294 hepaticus, 711 humidus, 56 hygroscopicus var. angustmy- ceticus, 1042 hygroscopicus, 45, 57, 235, 246, 1041, 1083, 1111, 1112, 1139, 1174, 1192 hygrostaticus n. sp., 1175 K-300, 894 kanamyceticus, 52, 53 kentuckensis , 1261 kitasatoensis, 275 kitazawaensis, 848 lactis, 1145 lavendulae, 731, 733-736, 894, 899, 1100, 1223 lavendulae -xesevnbMng, IIQ, 774 limosus, 224 iipwanzi-resembling, 1295 lucensis, 228 luteochromogenes n. sp., 728 lydicus, 1279 mashuensis, 54 matensis, 822, 1201 mediocidicus n. sp., 245 mediterranean, 593 melanogenes, 849 melanosporus (sine melanos- porofaciens) n. sp., 1148, 1196 michiganensis , 335 misakiensis, 997 mitakaensis, 1204, 1205 nagasakiensii n. sp., 894 narboensis n. sp., 274 natalensis, 226 nayagaiwaensis n. sp., 1163 netropsis, 918 nitrosporeus , 257 niveoruber Ettlinger et aZ., 617 niveus, 885 noboritoensis, 58 nodosus, 248 noursei, 230, 308 noursei variant, 1241 n. sp., 240, 250, 1099 Streptomyces abikoensis olivaceus (Waksman) Waks- man and Henrici, 306, 440, 576, 744, 745, 931, 1290 olivochromogenes, 1310 orchidaceus, 894 orientalis n. sp., 1300 parvullus, 335, 794 parvus, 334 paucisporogenes, 64 penticus, 241 phaeochromogenes var. c/iZo- ronz2/ceficus-resembling, 1296 p^a/oc/?rornogenus-resembllng, 1215, 1260 phoenix, 1267 platensis, 1139 pleofaciens, 1246 plicatus, 1020-1022 polychromogenes, 671 puniceus, 727 purpurascens , 275, 1180 purpurochromogenes, 1154 purpurochromogenes- resembling, 1306 pyridomyceticus , 752 racemochromogenes n. sp., 790, 1258 ramulosus n. sp., 150 resistomycificus, 575 reticuli, 1270 reticuli var. latumcidus, 1183 reticidi var. aqiiamtjceticus, 5 rimosus, 231, 274, 275, 306, 610, 612, 670, 1139 rimosus form paromomycins, 59 roc/iei, 1122 roseochromogenes, 732, 931, 1265 roseoc^roTnogenes-resembling, 1275 roseodiastaticus , 924 roseofiavus, 1219 roseus, 1287 rwber, 1224 ruber (Krainsky, Waksman and Henrici), 924 771 Microorganism Index Streptomyces abikoensis rutgersetisis var. castelarense n. var.. 1124 rutgersensis-resemhhng, 1272 sahachiroi, 1127. 1128 sakaieusis, 12 Hi siudeuensis, 1022 sp., 8. 40. 222, 141, 229, 232, 233, 247. 2(iO. 261, 275, 280, 281, 286-289, 310. 334, 335, 439, 440. 594, 611. 613-616, 621, 634. 689. 729. 742, 753, 755. 769, 796-801, 831. 1065, 1071, 1077, 1081, 1086-1089, 1101, 1103, 1104, 1110. 1129, 1134, 1154, 1158, 1167, 1188, 1193-1195, 1217, 1221, 1231, 1239, 1240, 1262, 1264, 1278, 1288, 1313. 1314 sp. No. 7017, 1082 sp. No. 14420, 1162 sp. PRL 1642, 767 sp. -resembling S. fradiae, 62 sp. -resembling S. lavendulae, 63 spectahilis, 1280-1284 spheroides, 885 spp., 60, 253-255, 604-606, 737, 914, 1064, 1076, 1228 strain No. 4738, 1141 subtropicus, 765, 766 tanaschieiisis related to s. a7i- tibioticiis, 577 tanaschiensis type, 1161 thioluteus, 870, 995 toyocaensis, 1298 vendargensis, 1304 venezuelae, 626 verticillaUis, 1305 verticillis, 1243 vinaceus, 727 vinaceus-drappus, 1022, 1080 violaceaniger, 1225 violaceus, 834, 1307 virginiae, 899 viridoflavus , 252 viridosporus, 234 xanthochromogenes n. sp., 1312 xanthophaeus, n. sp., 773 Streptomyces abikoensis zaomyceticus, 249 zaomyceticus n. sp., 835 Streptomycctaccac family, 118 Streptomycetes, 190, 212, 225, 237, 261, 262, 332, 334, 434, 436, 492, 501,502, 678, 1046, 1075, 1184, 1239, 1313 Streptosporangium genus, 118 Stropharia cubensis Earle, 937 Teloschistes exilis Wainio, 555 flavicans (Sw.) Norm., 445, 555, 557 Thamnolia subvermicularis Asahina, 461, 462 vermicularis (Sw.) Schaer. 458 Thelephora palmata, 507 spp., 507 Thermoactinomyces genus, 118 Thiocystis violacea, 930 Tilletia laevis, 643 tritici, 643 Torula mellis, 142 utilis, 351,513, 515,900 Torulopsis sp., 50 utilis, 537 Trachypus scaber, 652 versipellis, 638 Trametes cinnabarina (Jacq.) Fr., 1001 odorata (Wulf) Fr., 349 suavolens (Finn.) Fr., 619, 622 Treponema spp., 933 Trichoderma viride, 302, 938, 1308, 1309 Tricholoma nudum, 641, 1230 Trichosporon capitatum, 142 Trichothecium roseum (Link), J 59, 327-330 Umbilicaria pustulata L. Hoffm., 22, 39, 475 Urceolaria cretacea, 365 Usnea barbata, 452 diffracta Wain., 467 Pfizer Handbook of Microbial Metabolites 772 Usnea japonica Wain., 447 jesoensis Asahina, 446 longissima Ach., 464, 467 sp., 860 Ustilaginales spp., 127 Ustilago maydis, 301, 302, 643, 695, 896 sp., 38 sphaerogena, 94 zeae, 83, 127, 412 Ustulina vulgaricus, 28 vulgaris, 80 Variola amara (Ach.), 437 Verticilliiim alho-atriim, 1 psalliotae, 501 Vibrio adaptatus, 171, 173 Volucrispora aurantiaca, 503 Wheat rust, 304 Xanthoria fallax (Hepp.) Arn., 548, 555, 557 parietina (L. ) Th. Fr., 554 parietina (L.) Beltram, 555 Xerocomus hadius, 641 sanguineus, 652 suhtomentosus, 652 Yeasts, 13, 15, 17, 17, 19, 30, 43, 50, 53, 77, 96, 99, 134-136, i54, J57, 159, 308, 310, 314-316, 318, 331-333, 335, 336, 340-344, 351, 352, 422, 426, 436, 447, 480, 496, 508-512, 525, 528, 532, 534, 548, 618, 655, 660, 686, 716, 718, 721, 858, 900, 903, 904, 912, 926, 927, 933, 934, 969, 970, 974, 975, 976, 1009- 1012. 1013, 1016, 1018, 1028, 1030, 1034-1040, 1045, 1046, 1051, 1057-1060, 1062, 1190