BOSTON PUBLIC LIBRARY

3 9999J63i7W9 jre SURVEY ON

GENERAL AND COMPARATIVE
ENZYME BIOCHEMISTRY OF BIRDS

Boston Public Library

SEP 2 4 1971

DEPOSITORY

United States Department off the Interior

Fish and Wildlife Service

Bureau of Sport Fisheries and Wildlife

Special Scientific Report-Wildlife No. 143

UNITED STATES DEPARTMENT OF THE INTERIOR
Fish and Wildlife Service
Bureau of Sport Fisheries and Wildlife

LITERATURE SURVEY ON GENERAL AND COMPARATIVE
ENZYME BIOCHEMISTRY OF BIRDS

by

H. Ping Pan, Biochemist

Gainesville Field Station

Patuxent Wildlife Research Center

Division of Wildlife Research

Special Scientific Report — Wildlife No,
Washington, D. C. • 1971

143

For sale by the Superintendent of Documents, U.S. Government Printing Office

Washington, D.C. 20402 - Price 70 cents

Stock Number 2410-0273

Table of Contents

Page

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INTRODUCTION ................ « a. 1

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I. Enzymes Identified in Studies with Birds « ••<.<, 0 . o o * . 1

1. Oxidoreductases 0#00090oeo«ooooo««oo« 1

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II e Miscellaneous Enzymatic Reactions 0 B 00 0 eo 0 <».<.<, » » 38

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APPENDIX A - Abbreviations in this report ...... ....... 62

APPENDIX B - Enzymes cited in this report (with recom-
mended nomenclatures and reactions) .......... 64

111

ABSTRACT

Available information on the enzyme biochemistry of birds is pre-
sented. A literature search revealed that birds differ mainly from
other animals in metabolism involving nitrogen elimination. Comparative
differences in enzyme biochemistry of only a few species of birds are
known,, Very little work has been done in the field of comparative
enzyme biochemistry of birds, and practically none on blackbirds „

IV

INTRODUCTION

Because information on the enzyme biochemistry, especially compara-
tive enzyme biochemistry of birds, will be very helpful in basic studies
of enzyme systems in nuisance birds, a survey was made of the available
literature on enzymes, enzyme systems, and metabolism of domestic and
wild birds, on general and comparative enzyme biochemistry, and on
related techniques and procedures used in these studies.

This survey has relied heavily on the literature available in the
libraries of the University of Florida, especially the Medical College
Library. The libraries of the University of California at Berkeley and
the University of Wisconsin at Madison also were visited. The Biological
Sciences Library of the former and the Medical College Library of the
latter were especially helpful. The survey was intended to cover all
pertinent literature available, complete papers or abstracts, up to the
spring of 1967.

The enzymes appearing in the literature are presented here in the
six main classes as recommended in the Report of the Commission on Enzymes,
International Union of Biochemistry (1965). The long-established trivial
names of the enzymes are used because the systematic names recommended by
the Commission are not yet in wide use. The six main classes of enzymes
are given in the following order: 1. oxidoreductases, 2. transferases,
3. hydrolases, 4. lyases, 5. isomerases, and 6. ligases. Whenever feasible,
the enzymes are presented in the numerical order of their respective
Enzyme Classification Number and described under their source tissues.
When more than one enzyme is mentioned in an article, the work is described
only under one single enzyme as a matter of convenience; the rest of the
enzymes are mentioned in their respective groups with reference to the
described enzyme for details. The pertinent biochemistry is described with
the enzymes. This survey also reports, under the title "Miscellaneous
Enzymatic Reactions," those works in which some enzymatic reactions were
mentioned but the enzyme or the enzyme systems were not specified.
Finally, a table of the abbreviations used, and a table giving the trivial
names of the enzymes surveyed, together with their recommended names and
reactions, are contained in Appendices A and B, respectively.

ENZYMES SURVEYED
I. Enzymes Identified in Studies with Birds

1. Oxidoreductases

This class of enzymes catalyzes oxidoreduction reactions. It
includes dehydrogenases, oxidases, peroxidases, and oxygenases.

(1) Alcohol dehydrogenase

Eye tissue

The activities of alcohol dehydrogenase, lactate dehydrogenase,
and malate dehydrogenase were found by Lukashevich (1964) to
localize in the mitochondria of ellipsoids of the rods and cones
of the retina of pigeons and other animals. In all the verte-
brates studied the mitochondria of rods and cones located in the
region of the ellipsoids showed a high endogenous dehydrogenase
activity and high ADH, LDH, and MDH activities. Exposure of the
retinas to light altered these activities in a manner that
varied depending on the activity involved, the type of photo-
receptor with which the mitochondria were associated, and the
species of vertebrate. Because of the location of the enzymes
and the fact that their activity was modified by the action of
light, it was suggested that these three enzymes must partici-
pate in transmission of light stimuli from the outer to the
inner part of photoreceptors.

Also see Chilson et al. (1966) — lactic dehydrogenase, p. 5.

(2) D-3-Phosphoglycerate dehydrogenase

Chicken liver

Willis and Sallach (1964) partially purified D-3-phosphoglycerate
dehydrogenase from chicken liver and compared its distribution
to D-glycerate dehydrogenase. They revealed a marked variation
in the ratio of the activity of the first enzyme to that of the
second enzyme among animals studied, and that birds had the
highest levels of D-phosphoglycerate dehydrogenase. The enzyme
was further purified by Walsh and Sallach (1965) and its prop-
erties determined. It was inhibited by N-ethylmaleimide ,
3-bromopyruvate , and organic mercurials. Preincubation of the
enzyme with NADH or NAD+, but not with phosphoglycerate , partially
protected it against the first two reagents.

Also see Chilson et al. (1966) — lactic dehydrogenase, p. 5.

(3) Glucurono lac tone reductase

Bulbul (Ayes)

It is the absence of the microsomal enzymes (glucuronolactone
reductase and uronolactonase) that prevents the successful syn-
thesis of ascorbic acid in primates, ...and the bulbul (Reithel,
1967).

(4) Quinine dehydrogenase

Chicken kidney

The presence of oxidases of quinine in the chicken kidney,
but not in the chicken liver, was reported. The enzymes in
birds oxidized quinidine more rapidly than quinine, and the
reverse was true for the enzyme of mammals (Berheim, 1952).

(5) Malic oxidase

Chicken blood

Hunter and Hunter (1957) in their comparative study of
erythrocyte metabolism measured the oxygen consumption of
whole cells and the activity of malic oxidase, succinoxidase,
malic dehydrogenase, and lactic dehydrogenase of the homo-
genates of erythrocytes of chicken and other animals. They
found that at a given temperature, the nucleated red blood
cells of lower vertebrates, including birds, respire at a
level considerably above that of the non-nucleated, mammalian
erythrocytes. The succinoxidase activity of homogenates of
chicken erythrocytes and the malic oxidase activity of homo-
genates of chicken, fish, and some sting rays are considerably
higher than in other erythrocytes studied. The difference in
activity cannot be correlated with the presence or absence of
a nucleus.

Also see George and Talesara (1961c) — cytochrome oxidase, p. 14.

(6) Lactate dehydrogenase

a. Chicken heart, liver, and muscle

Wilson et al. (1963), compared the electrophoretic forms of
starch gel, the (H) and (M) types, and the hybrid types of
lactic dehydrogenase of the breast muscle of 38 species of wild
birds and domestic duck and fowl with the pure (H) and (M)
domestic fowl. Fondy et al. (1964), studied the comparative
enzymology of lactic dehydrogenases, and isolated and crystal-
lized the hybrid lactic dehydrogenases H2M2 from chicken liver
and HM2 from chicken leg muscle. The pure H4 form of the
enzyme had been isolated and crystallized from chicken liver
and compared to the H4 obtained from the heart. The fingerprint
patterns of tryptic digests of the hybrids, the immunological
properties, analog ratios, and oxalate inhibition were demon-
strated. McKay and Kaplan (1964) determined the protein
fluorescence polarization and the quantum yield of fluorescence
per mold or tryptophan for the heart (H) and muscle (M) types
of lactate dehydrogenases from chicken and beef. They found

that the (M) type had higher quantum yield than the (H) type.
They also found increases in quantum yields of fluorescence on
binding of NADH and the reduced 3-acetylpyridine analog of
NADH for each of the enzymes. They suggested that the
coenzymes might be bound closely to each other and might inter-
act only with certain tryptophan residues in the enzyme. Pesce
et al. (1964) reported methods for isolating and crystallizing
in pure form the chicken and beef skeletal muscle (M) lactic
dehydrogenase as well as chicken and beef heart (H) enzymes.
They also characterized the enzymes. The four lactic dehydro-
genases they isolated had somewhat similar amino acid
compositions. Most striking differences had been found with
the chicken muscle enzyme which contained approximately twice
as many histidine residues as the other enzymes. The chicken
muscle enzyme also contains significantly less glutamic acid
and tyrosine. The phenylalanine content of the two (H) type
enzymes is lower than that of the two (M) type enzymes. A
greater difference in amino acid composition between the chicken
heart and chicken muscle enzymes had been found than between the
beef heart and beef muscle enzymes. The authors discussed the
differences in properties of the enzymes with respect to the
enzymes' evolutionary and structural significances.

Schapira and Dreyfus (1964) made a comparative study of lactic
dehydrogenase isozymes of the skeletal muscle of the chicken.
They found that there are marked differences between the
embryonic and the adult muscle. In muscular atrophy and
dystrophy the isozyme pattern resembles the embryonic pattern.
The polymorphism of lactate dehydrogenase isozymes in wild,
racing, and pure-bred pigeons was investigated by Zinkham et
al. (1965). The isozyme patterns were found to be different.

The effects of temperature and substrate concentration on chick
lactate dehydrogenase activity were investigated by Maisel and
Kerrigan (1966). They showed by spectroanalysis that LDH in
breast muscle supernatant and isolated LDH 5 had greater activity
at high pyruvate concentration than at low. The difference was
more marked at 40°C than at lower temperatures. Heart muscle
LDH, and isolated LDH 1 reacted best at low pyruvate concen-
tration. The difference was less marked at 40°C than at lower
temperatures.

b. Chicken heart

Di Sabato and Kaplan (1964) studied the dissociation of lactic
dehydrogenases from chicken heart and beef heart into subunits
with sodium dodecyl sulfate. The dissociation of the enzymes
into subunits was accompanied by a loss of enzymatic activity.
They found that DPNH and its acetylpyridine analog protected
the enzymes from dissociation. The coenzymes also protected

against loss of catalytic activity. Cahn (1964) reported that
the lactate dehydrogenases isolated for chicken heart and from
breast muscle differ markedly in kinetics, ability to react
with coenzyme analogs, and electrophoretic mobilities. During
embryonic development, the enzyme content of the breast muscle
shifted from all heart type at early stages through intermediate
enzyme levels and types to all muscle type shortly after hatching.
Di Sabato (1965) showed that about 10 of the 30 tyrosyl residues
of chicken heart lactic dehydrogenase were free to titrate
between pH 8 and 11. He also showed evidence that the tyrosine
residues of the enzyme form a part of the site binding of the
coenzymes NAD. Fondy et al. (1965) made a further comparative
study of lactic dehydrogenases isolated from 19 species. The
sequence of the essential thiol peptide as isolated from chicken
H4 form of LDH was determined. This active site thiol peptide
from the LDH's may be functionally related to the active site
thiol region of the alcohol dehydrogenases.

c. Chicken embryo

Some cells in tissue culture and some tissues in chicken embryos
were used in the study of the rate of synthesis of lactic
dehydrogenase. It was found that it was regulated by the con-
centration of oxygen in the environment of the cells and the
tissues. This was particularly true with the synthesis of the
"muscle-type" of subunits of the enzyme (Goodfriend et al.,
1966). Lindy and Rajasalmi (1966) showed that incubation of
chick embryos in an hypoxic environment caused an increase in
the proportion of tissue lactate dehydrogenase made up of sub-
unit (H) , whereas incubation in aerobic conditions decreased
the proportional amount of subunit (M) . The variation of
ambient oxygen tension did not change the total lactate
dehydrogenase activity. They believed that these results
supported the hypothesis that oxygen or oxidative metabolites
had an effect on the synthesis of the subunit peptides.

Chilson et al. (1966) investigated the dissociation properties
and the reversible inactivation of chicken lactic and malic
dehydrogenases. They found evidence for the existence of an
intermediate form of lactic dehydrogenase with catalytic
properties altered during reactivation. Comparison also was
made with the dissociation properties of triosephosphate
dehydrogenase, rabbit muscle a-glycerophosphate dehydrogenase,
and horse liver alcohol dehydrogenase.

d. Gull nasal salt gland

McFarland et al. (1965) assayed the enzymes in the salt gland
of adult western gulls. The average units of enzyme activity
per gram of salt gland were as follows: lactic dehydrogenase,

90.1; phosphoglucomutase, 0.62; glucose-6-phosphate dehydro-
genase, 1.40; aldolase, 2.86; isocitric dehydrogenase, 5.08;
malic enzyme, 0.92; glutamic-oxaloacetic transaminase, 100.5;
and glutamic-pyruvic transaminase, 0.50.

e. Chicken eye

Maisel et al. (1965) analyzed the ontogeny of LDH isozymes in
the lens of the White Leghorn chick. They were able to show
the progressive appearance of nine molecular forms.

f . Penguin

Stability of lactate dehydrogenase isozyme patterns in penguins
was investigated by Markert and Sladen (1966). Three species
were used. The starch-gel zymograms of penguins were found to
be unusual in that the net charge on the isozymes is more
positive than is true for mammals or other birds that have been
investigated. In the adult, isozyme content of tissues were
highly constant. The heart tissue pattern was the same as that
of a fish. The zymograms prepared from various tissues of
Merluccius bilinearis differed.

g. Peafowl

The multiple forms of lactate dehydrogenase of peafowl were
studied by Rose and Wilson (1966). The electrophoretic
properties of the peafowl isozymes were found to be unusual in
that the isozyme from heart tissue can be either more or less
anodic than that of muscle, depending on the pH.

Also see Lukashevich (1964) —alcohol dehydrogenase, p. 2;
Hunter and Hunter (1957)— malic oxidase, p. 3;
George and Talesara (1961c) — cytochrome oxidase, p. 14;
McDaniel and Chute (1961) — glutamic oxaloacetic
transaminase, Class 2, p. 18.

(7) Malic dehydrogenase

a. Chick embryo

Novikoff et al. (1948) showed the presence of DPN in the malic
dehydrogenase system of early chick embryo.

b. Pigeon liver

Hsu and Lardy (1967a) prepared malate dehydrogenase from
pigeon liver, purified it, and crystallized it as its
triphosphopyridine nucleotide complex. The properties of the
crystal were determined. The purified enzyme catalyzed the

decarboxylation of oxalacetate at a rate comparable to the
decarboxylation of malate. Reversible inactivation of these
activities occurred upon storage of the enzyme. Reactivation
was obtained by incubation with dithiothreitol. They later
studied (1967b) the fluorescence of the coenzyme binding of
the purified enzyme, which fluoresced strongly with excitation
and emission maxima at 293 and 350 mji. Upon binding with TPNH,
a blue shift in the emission maximum of TPNH from 465 to 440
trai, accompanied by an enhancement of fluorescence intensity,
takes place. The binding of the enzyme with L-malate or TPNH
was specific, and was not observed with either D-malate or
reduced diphosphopyridine nucleotide.

c. Chicken heart

Kitto and Kaplan (1966) isolated and purified the mitochondrial
and supernatant forms of malic dehydrogenase from chicken heart.
The supernatant and mitochondrial enzymes differed markedly in
their amino acid composition and their peptide maps. Other
comparative properties also were reported.

d. Enzyme evolution studies with malate dehydrogenase

Kitto and Wilson (1966) subjected heart extracts from over 100
species of birds to starch-gel electrophoresis. The "super-
natant" form of malate dehydrogenase, which was present in
every extract, was located on the gels. The mobility of this
enzyme showed very little interspecific variation. Most birds
tested had a supernatant malate dehydrogenase that moved as
fast as the chicken enzyme. Hummingbirds and swifts, however,
which were usually considered as two suborders of Apodiformes,
were unique among the birds tested in having an enzyme that
moved slower than the chicken enzyme. This finding appeared to
confirm, they believed, the unity of the Apodiformes, an order
whose unity had long been open to question. All families of
the shorebird order (Charadriiformes) tested were unique in
having an enzyme that moved at a definite distance from the
chicken enzyme. The unity of this order of birds was also
previously open to question.

Also see Lukashevich (1964) — alcohol dehydrogenase, p. 2;
Hunter and Hunter (1957) — malic oxidase, p. 3;
Chilson et al. (1966) —lactate dehydrogenase, p. 5;
McFarland et al. (1965) — lactate dehydrogenase, p. 5-6;
McDaniel and Chute (1961) — glutomic oxaloacetic

transaminase, Class 2, p. 18;
Rubinstein and Denstedt (1953) — citrate-cleavage

enzyme, Class 4, p. 34;
Goodridge and Ball (1966) — acetyl CoA carboxylase,

Class 6, p. 37-38.

(8) 6-Phosphategluconic dehydrogenase

Chicken blood

Salvidio et al. (1963) in their study of the glucose-6-
phosphate and 6-phosphategluconic dehydrogenases activities
in the red blood cells of several animal species, showed that
6-phosphategluconic dehydrogenase activity in pigeon red
blood cells had an intermediate value of positivity, higher
than that of human.

Also see Goodridge and Ball (1966) — acetyl CoA carboxylase,
Class 6, p. 37-38.

(9) Glucose -6 -phosphate dehydrogenase

a. Chick embryo

The glucose-6-phosphate dehydrogenase activity was found in
the homogenates of chorioallantoic membranes of the chick
embryo. This enzymatic activity recovered in the centrifugal
supernatant fluid did not change appreciably with age of the
embryo between 6 and 13 days (Kun, 1953). The activity of
this enzyme in the developing chick embryo was found to
localize primarily in skin and feather primordia of appendages
from older embryo (Mahaffey, 1961).

b. Chick spinal cord

Burt and Wenger (1962) and Burt (1963) determined the activity
of this enzyme in the spinal cord of the developing chick as
an index of pentose cycle function during embryogenesis of the
brachial chord. Their finding, they believed, supported the
postulate that following cellular proliferation an early step
in mural differentiation consisted of a transient rise in
pentose cycle activity.

c. Chick embryo

Newburgh et al. (1962) used chick embryo explants in their
study of the relation of enzymes glucose-6-phosphate dehydro-
genase and isocitric dehydrogenase to protein and DNA
accumulation. They found a much closer correlation of activities
of the two enzymes with changes in DNA content than in the total
protein content.

Also see McFarland et al. (1965) — lactate dehydrogenase, p. 5-6;
Goodridge and Ball (1966) — acetyl CoA carboxylase,
Class 6, p. 37-38.

(10) Triosephosphate dehydrogenases

Turkey breast muscle

Turkey and other animals were used in the study of the effect
of tetrathionate on the stability and immunological properties
of muscle triosephosphate dehydrogenases. When the three
sulfhydryl groups of the enzymes were modified by tetrathionate,
the enzymes were completely inhibited. Depending on the tem-
perature and length of incubation with the inhibitor, this
inhibition could be reversed by the addition of thiols. While
the immunological properties of the inactivated turkey TPD
differed significantly from the native enzyme, the immunological
properties of the sturgeon enzyme were only slightly altered
(Allison and Kaplan, 1964). A comparative study was made on
the enzymology of triosephosphate dehydrogenases from chicken,
pheasant, and turkey breast muscles, and those from the human
breast, the skeletal muscle, and the tail muscle of lobster.
Their physical, chemical, and immunological properties were
compared with those of rabbit muscle and yeast (Allison, 1964).

Also see Chilson et al. (1966) — lactate dehydrogenase, p. 5;
Rinaudo (1962) — hexosephosphate isomerase, Class 5,
p. 36.

(11) Pes tradiol-17B- dehydrogenase

Chicken and pigeon blood

Portius and Repke (1960) demonstrated the presence of oestra-
diol-17|8-dehydrogenase in the blood of pigeon and hen and
other vertebrates. However, sheep, goat, and cow contained
oestradiol-17a-dehydrogenase.

(12) Xanthine oxidase

a. Pigeon liver and kidney

Two tissues were necessary for uric acid synthesis in pigeon.
The primary step, the binding of ammonia with some uncertain
source of carbon, occurred in the liver where hypoxanthine
was formed. The final conversion into uric acid, which took
place in the kidney (or pancreas), was an oxidation catalyzed
by xanthine oxidase (Edson et al., 1936). Goodwin (1960),
however, reported that pigeon liver did not contain xanthine
oxidase and thus did not oxidize to uric acid.

b. Chicken tissue

Croisille (1964) showed that hypoxanthine dehydrogenase
(xanthine oxidase) was synthesized in different tissues at
three very different periods during development of the chick.

9

The enzyme was almost specifically localized in the kidney
during embryonic life; after hatching, it was present mainly
in the liver. The enzymatic activity in the different tissues
of the chick could be attributed to the presence of two very
closely related proteins — one present in' the mesonephros and
the definitive kidney; the other present in the liver, the
intestine, and the pancreas. The sites responsible for the
antigenic and enzymatic properties of the molecule appeared to
be different.

(13) Xanthine dehydrogenase

a. Chick liver

De Angelis and Totter (1964) studied the kinetics of hydrogen
transfer between hypoxanthine and dimethylbiacridylium nitrate
in the presence of chick liver xanthine dehydrogenase. They
found that a two electron transfer from hypoxanthine to
10,10'-dimethyl-9,9'-biacridylium ions (DBA"*"*") , followed by
reoxidation of the reduced DBA++ by oxygen, could account
largely for the initial velocity of uric acid production cata-
lyzed by chick liver xanthine dehydrogenase in the presence of
DBA"1""*". The sensitivity of the chain reaction was discussed.
Stirpe and Corte (1965) reported that xanthine dehydrogenase
activity of chick liver increased during starvation. Adminis-
tration of inosine, and possibly of adenine, had a comparable
effect on the xanthine dehydrogenase, and also induced an
elevation of the total quantity of enzyme. Hypoxanthine,
xanthine, guanine, xanthosine, guanosine, and adenosine were
ineffective. Cortisone was equally ineffective. The adminis-
tration of puromycin abolished the effect of inosine and
reduced that of starvation. Inosine induced increased synthesis
of xanthine dehydrogenase, whereas during starvation the enzyme
was spared with respect' to other liver proteins. The authors
suggested that chick liver xanthine dehydrogenase is an adaptive
enzyme.

b. Chicken kidney

Cardona et al. (1965) studied in vitro the enzyme system xan-
thine dehydrogenase (XDH) in chicken tissue utilizing tissue
homogenates, xanthine (sodium salt) as substrate, and methylene
blue as hydrogen acceptor. Optimum conditions were determined
for the enzymatic activity. Kinetic studies also were conducted
with inhibitor and activators.

(14) Succinic dehydrogenase

a. Pigeon muscle

The relative quantitative distribution pattern of the narrow
red and broad white fibers, and the succinic dehydrogenase

10

activity in the different layers of the pigeon breast muscle
were studied quantitatively. The activity of this enzyme in
any particular layer of the muscle was related to the number
of the narrow red fibers present as the main bulk of the enzyme
resided in these fibers (George and Talesara, 1960). This
enzyme and cytochrome oxidase activity in pigeon muscle were
studied by means of histological methods. Succinic dehydro-
genase was detected in broad white fibers. The presence of a
large number of succinoxidase-positive granules in the narrow
red fibers accounted for their high oxidative metabolism. The
presence of fewer, and smaller, succinoxidase-positive granules
in the broad white fibers accounted for their smaller oxidative
activity. The granules were suggested to be mitochondria
(George and Talesara, 1961a). Later the study on succinic
dehydrogenase included a few representative types of birds and
a bat. In all the birds and the bat the pectoralis major
muscle showed higher activity than the pectoralis minor, which
indicated that of the two, the former muscle was metabolically
much active and highly evolved. The pectoralis major muscle
of the rosypastor, sparrow, bat, parakeet, and pigeon showed
higher concentrations of succinic dehydrogenase as compared to
the other birds studied whereas the fowl, a nonflier, showed
the lowest enzyme concentration. In the birds studied a recip-
rocal relationship was found between the enzyme activity of the
pectoralis major muscle and the body weight, fiber diameter,
and weight of the pectoralis muscle (George and Talesara, 1961b)

Succinic dehydrogenase in pigeon pectoralis during disuse
atrophy was investigated by Cherian et al. (1965). Muscle
enzyme activity decreased from the first day, reaching its low-
est level after 7 days. Histochemical observations on red and
white muscle fibers showed that after 7 day's atrophy the
majority of red fibers had lower enzyme activity; a few others
showed higher activity. In white fibers, on the other hand, a
uniform increase over normal in enzyme activity and mitochon-
drial number was noticed. These changes in the two types of
fibers were less conspicuous during the later stages of atrophy.
The authors suggested that in both red and white fibers there
was a shift from aerobic to anerobic metabolism, or vice versa.
Histochemical study of oxidative enzymes showed that the
striated muscle fibers of various kinds of vertebrate muscles
fell into three types — small red muscle fibers which showed
higher activities, large white fibers with lower activities,
and "medium fibers" with intermediate activities. The chicken
muscle was composed of three types of fibers, while pigeon and
lovebird muscles displayed two types of fibers (Ogata and Mori,
1964).

11

b. Pigeon taste bud

Pevzner (1964) found succinic dehydrogenase activity to be
localized in the taste buds of pigeon and other vertebrates.
The author also observed that the localization and degree of
enzymatic activity in mitochondria of the taste buds remained
essentially constant for the animals on various steps of the
evolutionary ladder.

c. Pigeon heart

Lee (1963) prepared pigeon heart mitochondria by sonication
and also used digitonin to treat particles from pigeon heart.
He found that the relative values of ATPase : NADH oxidase :
succinate oxidase : energy-linked NAD reductase : energy-
linked ferrocytochrome c oxidase in mjimole/min/mg protein were
1180:470:31:40:2.4 for sonicated preparation and 230:190:14:
5.5:12 for digitonin-treated particles.

d. Chicken embryo

Masai et al. (1965) localized the succinic dehydrogenase and
manoamino oxidase activity in the central nervous system of
the chick embryo by means of a histochemical method. They
postulated that the neural tube was subdivided into columns.

Also see Hunter and Hunter (1957) — malic oxidase, p. 3;

George and Vallyathan (1964) — lipase, Class 3, p. 21;
Rubinstein and Denstedt (1953) —citrate-cleavage
enzyme, Class 4, p. 34.

(15) Glutamate dehydrogenase

Chicken and pigeon liver

Frieden (1965) isolated glutamate dehydrogenases from animal
and nonanimal sources, and used the beef liver enzyme for
comparison for effects of purine nucleotides on their prop-
erties. The enzymes from all animal sources were strongly
and specifically affected by purine nucleotides. The binding
constants and extent of activation or inhibition differed
with different enzymes. However, the enzymes from pigeon and
chicken livers appeared identical and they were different from
those from mammalian livers.

(16) Folic acid reductase

Chicken liver

Folic acid reductase from chicken liver was used by Rothenberg
(1965) to reduce the tritiated folic acid to tetrahydrofolic

12

acid with reduced triphosphopyridine nucleotide as the cof actor.
From this he developed a method for determining methotrexate
levels in biological fluids.

Also see Matsubara and Akino (1964) — sepiapterin reductase, p. 13.

(17) Dihydrofolic reductase

Chicken liver

Mathews and Huennekens (1963) highly purified dihydrofolic
reductase from chicken liver. The enzyme was characterized, and
its reaction reversibility demonstrated. Kaufman (1963) found
that under a variety of conditions the activity of dihydrofolic
reductase was stimulated in a qualitatively similar manner by
salts, urea, formamide, and possibly hydrogen ions.

Also see Matsubara and Akino (1964) — sepiapterin reductase, p. 13.

(18) Sepiapterin reductase
Chicken liver

The presence of this enzyme in chicken was reported by Matsubara
and Akino (1964).

(19) Cystine reductase

Chicken liver

Cazorla and Barron (1958) showed that the embryonic liver of
chicken contained cystine reductase. The enzyme reduced cystine
to cysteine with DPNH as the specific coenzyme. TPNH was in-
effective. They also found that glutathione reductase, TPNH-
specific, was also present.

(20) Uricase

Chicken embryo

Prosser and Brown (1961) in their description of uricolytic
enzymes, stated that uricase activity in chick embryo increased
up to 10 days only.

(21) Glutathione reductase

Chicken liver

The activity of the enzyme glutathione reductase was signifi-
cantly lower in the liver preparations from vitamin ^\2~
deficient chicks than in those from normal chicks (Biswas and
Johnson, 1964) .

Also see Cazorla and Barron (1958) — cystine reductase, p. 13.

13

(22) Catalase

Chicken blood

Foulkes and Lemberg (1949-1950) presented the comparative
amounts of catalase activities in these three species: horse,
1500; chicken, 86; and duck, 4.

Also see Cohen and Hochstein (1963) — glutathione peroxidase,
p. 14.

(2 3) Cytochrome oxidase

Pigeon muscle

George and Talesara (1961c) quantitatively studied the distri-
bution pattern of cytochrome oxidase, malic oxidase, succinoxi-
dase, lactic dehydrogenase, and lipase in the different layers
of pigeon breast muscle. The values derived for the individual
red as well as white fibers indicated that the bulk of all the
enzymes studied resided in the red fibers. These enzymes occur
in only very low concentrations in the white fibers.

Also see Lee (1963) — succinate dehydrogenase, p. 12;

George and Talesara (1960) — succinate dehydrogenase,
p. 10-11.

(24) Glutathione peroxidase

Duck blood

Catalase-def icient duck erythrocyte cells were protected against
the toxic effects of low hydrogen peroxide concentrations by
sustained glutathione peroxidase activity (Cohen and Hochstein,
1963).

(2 5) Cytochrome £ reductase

See Lee (1963) — succinate dehydrogenase, p. 12.
(2 6) Monoamine oxidase

See Aprison et al . (1964) — hydroxytryptophan decarboxylase,
Class 4, p. 34.

(27) Isocitric dehydrogenase

See McFarland et al. (1965) —lactate dehydrogenase, p. 5-6;

Newburgh et al. (1962) — glucose-6-phosphate dehydrogenase,

p. 8;
Rubinstein and Denstedt (1953) —citrate-change enzyme,

Class 4, p. 34.

14

(28) D-Glycerate dehydrogenase

See Willis and Sallach (1964) — D-3-phosphoglycerate dehydro-
genase, p. 2.

(29) Tryptophan pyrrolase

See Knox and Eppenberger (1966) —tyrosine transaminase, Class 2,
p. 18.

(30) B-Hydroxybutyrate dehydrogenase and

(31) Diaphorase

See Hellerstrom (1963) — alkaline and acid phosphatases, Class 3,
p. 24.

2. Transferases

This group of enzymes catalyzes group transfer. It includes trans-
aminases, kinases, and transacetylases.

(1) Catechol methyltransf erase

Chick embryo

The actions of enphephrine on the innervated embryonic chick
heart were terminated primarily through the metabolism by means
of catechol-O-methyltransferase (McCarty and Shideman, 1960).

(2) Ornithine carbamyl transferase

Chicken tissues

Siren (1963) studied the arginine metabolism in chicken.
Arginine is an essential amino acid for birds. The author, in
order to clarify whether chicken could synthesize arginine,
determined the activity of ornithine carbamyl transferase,
which is an enzyme in the urea cycle in different organs of
the chicken. The enzyme was found to be absent, and no inhibi-
tor was found. He concluded that chicken could not synthesize
arginine.

(3) Carnitine acetyltransf erase

Chicken breast muscle

Beenakkers and Klingenberg (1963) showed that pigeon breast
muscle had the highest activity of the enzyme carnitine acetyl -
transferase in mitochondria with acetylcarnitine as substrate
of any animals tested. This enzymatic activity later was found
also in pigeon breast muscle by Chase et al . (1965).

15

(4) Phosphorylase

Pigeon liver

The activity of phosphorylase in pigeon liver was higher as
compared to that of glycogen synthetase. While G6P activated
and UMP inhibited synthetase, G6P inhibited and UMP activated
phosphorylase. 2-Deoxy-G6P acted in the same way to G6P
(Fridland and Nigam, 1965).

(5) Glycogen phosphorylase

Chick embryo

Guha and Wegmann (1961) showed that ATP and Mg"1""^ were needed
in the histochemical demonstration of glycogen phosphorylase
in the chick-embryo liver. Grillo (1961) further localized
the enzyme in the liver parenchyma. It occurred in the liver
parenchyma from the seventh day onwards, in cardiac muscle
from the third day onwards, and in skeletal muscle from the
13th day onwards. The authors suggested that although
phosphorylase was important in glycogenolysis it could not
initiate the formation of glycogen in embryonic tissues.

(6) Uridine diphosphate glucose-glycogen synthetase

Chick embryo

Grillo and Ozone (1962) assayed UDPG-synthetase activities in
chick-embryo tissues. In heart, liver, skeletal muscle, and
brain, UDPG activities occurred earlier in development than
phosphorylase and paralleled the appearance of glycogen in
tissues. Glycogen synthesis occurred without insulin.

(7) Glutamine phosphoribosyl-pyrophosphate amidotransferase

a. Pigeon liver

Caskey et al . (1964) purified the enzyme from pigeon liver,
and conducted inhibition studies. They concluded that the
enzyme was susceptible to inhibitors only when in a specific
conformational state, and that inhibitors acted by distorting
the enzyme and reducing its activity.

b. Chicken liver

Hartman (1963a, 1963b) purified the enzyme from chicken liver,
and made it apparently homogenous in the ultracentrifuge. It
was inhibited by an analog of glutamine, and it had 10 iron
atoms per molecule. Chelation of the iron atoms inhibited the
enzymatic activity.

16

(8) Glucokinase

Pigeon muscle

Marcus and Manery (1966) studied the potassium stimulation of
oxidation and phosphorylation in pigeon-muscle mitochondria.
They used a-ketoglutarate as substrate and hexokinase — glucose
phosphate acceptor system. It was found that the rate of
phosphorylation was stimulated to an even greater extent than
the rate of oxidation as sodium was replaced by potassium. In
every preparation tested, the rates of phosphorylation and the
P/0 ratios were higher in the high-potassium media than in the
high-sodium media.

(9) NAD kinase

Pigeon liver

Nemchinskaya (1963) found an enzymatic system for NADP synthe-
sis from NAD and ATP in hyaloplasm of pigeon liver cells. The
NAD kinase was stable during initial purification for 1 month.

(10) Creatine kinase

Chicken brain, heart, and muscle

Dawson et al . (1965) showed that in chicken all muscle con-
tained only one creatine kinase, "muscle enzyme". Brain
contained exclusively "brain enzyme". Yeast contained "brain
enzyme" and might contain the intermediate enzyme. They
showed that creatine kinase was an enzyme whose structure was
that of a dimer.

Eppenberger et al. (1967) and Dawson et al. (1967) studied the
comparative enzymology of creatine kinases. They purified two
creatine kinases from chicken brain and muscle and compared
them to those from the rabbit. There were significant dif-
ferences in amino acid composition between the brain and
muscle types; the hybrid enzyme had an intermediate composition.
Antibodies prepared against the chicken muscle type enzyme did
not cross-react with the chicken brain type enzyme and the
rabbit muscle type enzyme. However, the hybrid enzyme reacted
with both antibodies. The two brain type enzymes from the
chicken and rabbit, or the two muscle enzymes, were more
similar in their properties than were the brain and muscle
forms of one species. The physical and chemical properties
of the enzymes were determined.

17

(11) Glutamic oxaloacetic transaminase and

(12) Glutamic pyruvic transaminase

Chicken blood

McDaniel and Chute (1961) measured the levels of enzymes in
two breeds of chicken. The enzymes measured included glutamic
oxaloacetic transaminase, glutamic pyruvic transaminase, acid
phosphatase, lactic dehydrogenase, malic dehydrogenase, and
alkaline phosphatase.

Also see Cornelius et al. (1959) — aldolase, Class 4, p. 34;
McFarland et al. (1965) — lactate dehydrogenase,
Class 1, p. 5-6.

(13) Tyrosine aminotransferase

a. Chicken liver

Litwack and Nemeth (1965) found that in the homogenate of
chicken liver the tyrosine aminotransferase activity was
relatively constant during the embryonic period. After
hatching, a somewhat variable 2- to 3-fold increase was ob-
served which persisted for approximately 10 days. The
activity in the homogenates of chicken liver obtained at
steps during embryonic life and after hatching was relatively
specific for ot-ketoglutarate.

b. Chicken embryo

Knox and Eppenberger (1966) showed that in the liver of chick
embryos tyrosine transaminase was present from the fifth day,
tryptophan pyrrolase from the 11th day, and formylase appeared
at the time of hatching. The tyrosine transaminase level was
elevated 5-fold by treatment with hydrocortisone but not with
tryptophan, and the tryptophan pyrrolase was similarly
elevated by treatment with tryptophan but not with hydro-
cortisone. The developmental elevation of formylase was not
affected by either treatment. The behavior of the enzymes
was, therefore, markedly different in the chick embryo than
in the fetal rat.

(14) Phosphoglycerate mutase

Chick muscle

Phosphoglycerate mutase was prepared from commercially available
frozen chicken breast in high yield. The procedure for the
preparation and the properties of the enzyme were described by
Torralba and Grisolia (1966).

18

(15) Amidino transferase

Chick embryo

Walker and Wang (1964) determined the relationships between
liver-creatine concentration and L-arginine:glycine amidino-
transferase activity in the closed system of the developing
chick embryo. Creatine, or phosphocreatine, appeared to be
the specific metabolite-repressor of the enzyme.

(16) Hexokinase

Chick embryo

Guidotti et al. (1964) found that in 10-day chick embryo
hearts, free glucose accumulated at an extracellular concen-
tration exceeding 24 mM. Below this concentration the inward
transport of glucose conformed to a Michaelis-Menten kinetics
with hexokinase 1^=2.3X10 M and a transport maximum of about
7 mg/g wet wt/hr.

(17) Formylglycinamide ribotide amidotransferase

(18) 5 -Phosphor iboxylpyrophosphate amidotransferase

(19) Glycinamide ribotide transformylase and

(20) 5-Amino-4-imidazolecarboxamide ribotide transformylase

Chicken and pigeon liver

Flaks and Lukens (1963) in their review on the enzymes of
purine nucleotide synthesis de novo, stated that avian livers
have the highest known concentrations of purine synthetic
enzymes of any animal tissue studied, since the pathway was
utilized by birds as a means of nitrogen elimination.

(21) Transaldolase and

(22) Transketolase

See Goodridge and Ball (1966) — acetyl CoA carboxylase, Class 6,
p. 37-38.

(23) NAD pyrophophorylase

See Lee (1963) — succinate dehydrogenase, Class 1, p. 12.

(24) Choline acetylase

See Aprison et al. (1964) — hydroxytryptophan decarboxylase,
Class 4, p. 34.

19

(25) Phosphoglucomutase

See McFarland et al. (1965) — lactate dehydrogenase, Class 1,
p. 5-6.

(26) Myokinase

See Kitiyakara and Harman (1953) — ATPase, Class 3, p. 30.

3. Hydrolases

Enzymes of this group hydrolyze a variety of compounds. The fol-
lowing enzymes were found in the studies of birds.

(1) Aliesterases and

(2) Arylesterases

Chicken and duck blood

Electrophoresis studies on blood plasma esterases on a cellulose
column showed that chicken plasma were characterized by a
propionylcholinesterase. This was the only esterase present
and it differed from other cholinesterases in many respects.
Duck plasma contained a propionyl-p-esterase in addition to a
propionylcholinesterase (Augustinsson, 1959a). Further study
with 27 various vertebrates showed that arylesterase was the
predominant esterase in mammalian plasma, and was absent in
the plasma of chicken and duck, as well as being absent in
reptiles, amphibians, and fish (Augustinsson, 1959b).

Kaminski (1964) found that duck serum contained three esterases.
One of the enzymes was characterized as the cholinesterase and
two others as aliesterases.

(3) Lipase

a. Chick pancreas and small intestine

Lipase and esterase activities of the pancreas and small in-
testine of the chick were investigated. In chicks from 1 to
30 days of age the lipase activity of the pancreas remained
fairly constant, but the esterase activity of the small intes-
tine increased markedly from 1 to 10 days of age and then
remained constant. Of the three sections of the small intes-
tine examined, lipase activity was greatest in the upper small
intestine, whereas esterase activity was greatest in the duodenum.
Although lipase activity was almost entirely confined to the
pancreas, and esterase activity to the small intestine, evidence
was given that both enzymes were present in both tissues (Laws
and Moore, 1963).

20

b. Chick embryo

During the 8 to 19 days of incubation the lipase activity in
the liver was found to increase gradually from the eighth to
the 14th day, and then to shoot up suddenly, reaching the peak
on the 16th day. This rise in activity was correlated with
the extent of fat metabolism in the embryo during the period
(George and Iype, 1962).

c. Pigeon muscle

An improved histochemical demonstration of lipase activity with
pigeon breast muscle was described. The red narrow fibers
contained a large concentration of lipase, while the white,
broad ones did not. Tissue samples coated with gelatine
improved with histochemical test for the presence of lipase
(George and Iype, 1960). In the particulate fractions of the
pigeon breast muscle, the soluble fraction showed highest
lipase activity; mitochondrial and microsomal fractions showed
only one half the activity. The specific activity of lipase,
however, was found to be maximum in the microsomes (George and
Talesara, 1962a). The mitochondrial lipase and SDH activities
were strikingly low in the premigratory phase (George and
Vallyathan, 1964). Histochemically, lipase activity was demon-
strated by Bokdawala and George (1964) in pigeon breast muscle
using "Tween 85" as substrate and alizarin red S as stain for
the calcium soap formed.

Pope and Tidwell (1964) purified from homogenates of chicken
intestinal mucosa an enzyme catalyzing the hydrolysis of mono-
glycerides. It was liberated from lyophilized microsomes by
treatment with sodium deoxycholate and subsequently with
calcium chloride. Unlike pancreatic lipase, the activity did
not seem to be promoted by taurocholate.

(4) Choi inest erases

Chick blood

Mendel et al. (1943) in a test using acetyl-p-methylcholine,
which was hydrolyzed by true but not pseudo-cholinesterase and
benzoylcholine as substrates, found that the plasmas of chicks,
ducks, and pigeon contained true and pseudo-cholinesterases
(with pigeon not conclusive in pseudo-cholinesterase). There
was neither true nor pseudo-cholinesterase in their cells.

Also see Augustinsson (1959b) — aliesterases and arylesterases,
p. 20;
Plimmer and Rosedale (1922) — amylase, p. 27;
Leasure and Link (1940) — amylase, p. 30;
George and Talesara (1961c) — cytochrome oxidase,
Class 1, p. 14.

21

(5) Lactase

Chicken crop

Hamilton and Mitchell (1924) found lactase present in the crop
but absent in the proventriculus, the pancreas, and the intes-
tines of normal chickens.

Also see Plimmer and Rosedale (1922) — amylase, p. 27.

(6) Alkaline phosphatase and

(7) Acid phosphatase

a. Chicken blood

Laying hens had higher and more variable serum phosphatase
than cocks. No great difference in serum phosphatase could be
found between laying hens and hens in a period of suspended
egg production. Pullets from another source which had never
laid gave values comparable with the value for cocks. Serum
phosphatase was much higher in chicks suffering from rickets
(Common, 1936). The acid phosphatase activity was about equal
in the erythrocyte cells of birds and turtles, while alkaline
phosphatase activity in these cells was variable (Rapoport
et al., 1942). Stutts et al. (1957), who studied plasma alka-
line phosphatase activity in mature inbred chickens, believed
that differences in alkaline phosphatase activity in blood
indicated that the phosphatase level was in part genetically
controlled. In each line the enzyme activity of females was
higher and more variable than that of the males.

Tanabe and Wilcox (1960) found that serum alkaline phosphatase
in chickens was higher in most cases between 2 and 26 weeks of
age in a line bred for high alkaline phosphatase than in a
random bred control line. The level in the male was higher
than the female during the growing period, but lower at later
age. There were no differences at hatching due to the line or
sex. Bell (1960) found in his study with Brown Leghorns that
both adult nonlaying hens and cocks showed enzymatic activity
of the same order. The plasma of the mature birds (5 to 6 weeks
old) of both sexes had alkaline phosphatase levels up to 10X
those found in nonlaying adults. In the hen, the average level
of plasma enzymatic activity was increased by about 50 percent
when hens began to lay. The massive temporary increase in
plasma phosphatase shown by the laying hens was believed to
reflect a stimulation of the osteoblastic recalcif ication
process called into action by the drain on skeletal calcium
required for egg shells. Bell (1961) studied the plasma acid
phosphatase activity and bone dystrophies in the domestic fowl

22

and found that fowls with severe osteoporosis, where many
osteoclasts were present, usually showed an elevated plasma
acid phosphatase activity as well as greatly raised alkaline
phosphatase activity.

Dounce and Seibel (1943) found the enzyme acid phosphatase in
nuclei isolated from washed chicken erythrocytes. The activity
of fresh preparations determined by the use of disodium phenyl-
phosphate as substrate corresponded to the liberation of about
0.0025 to 0.0030 mg of phenol from the substrate per milligram
of tissue per hour at 25°C. Approximately the same concen-
tration of enzyme was found in the washed whole erythrocytes.
Hurwitz and Griminger (1961) conducted experiments to evaluate
the relationships between plasma alkaline phosphatase and plasma
inorganic phosphorus, bone constituents, and parathyroid size.
They found that calcium-deficient hens, irrespective of supple-
mentation with phosphorus, have enlarged parathyroids, elevated
plasma alkaline phosphatase, and loss of bone material. The
experimental results indicated that decalcification rather than
calcification of bone was associated with increased alkaline
phosphatase levels.

b. Red-winged blackbird brain

Rogers (1963a) used red-winged blackbird and chick (Gallus)
in his comparative biochemical alkaline phosphatase studies
in relation to brain development. The enzyme levels of red-
wing were determined in three brain parts during the 15 days
after hatching; those of chick were determined at the 19th
embryonic day, 6 days and 9 weeks after hatching. Redwing
enzyme increase in the posthatching period was similar to
that of chick brain in the late embryonic period. Enzyme
levels in both birds decline in adult after a peak was reached.
The author believed that the results supported the hypothesis
that alkaline phosphatase played a role in the differentiation
of the central nervous system. Rogers (1963b) also histo-
chemically localized the enzyme in the brains of chick, red-
wing and other mammals. He found that the results contained
data both for and against the association of alkaline phospa-
tase and myelination in the late development of the nervous
system.

Wilson and Wilcox (1963) determined the origin of the increased
serum alkaline phosphatase in chicks by means of enzyme
inhibition studies. Both high phosphatase line and random
bred line of chicks were investigated. Sodium borate was
found to rather selectively inhibit the serum phosphatase in
the high line. Further studies with this inhibitor on tissues
from both lines of chicks showed that intestinal phosphatase
activity was inhibited to a much greater degree than that

23

in bone, liver, and kidney. These studies indicated to the
authors that the major portion, if not all, of the increased
serum phosphatase of the high line originated in the intestine.

The influence of dietary minerals on the pH optima and reaction
velocities of phosphatases of the chicken was studied using
magnesium, calcium, and molybdenum ions (Motzok, 1963).

c. Duck pancreas

Enzyme histochemical study of the pancreatic islets in duck
showed that the islet B cells exhibited a strong reaction for
alkaline phosphatases, but a weak one for acid phosphatase.
The activities of G-6-Pase, ATPase, diphosphopyridine nucleotide
diaphorase, triphosphopyridine nucleotide diaphorase, and
g-hydroxybutyrate dehydrogenase also were positive. Amino-
peptidase and esterase activities were not found in the B cells,
however (Hellerstrom, 1963).

d. Chick embryo

Moog and Richardson (1955) showed that the duodenum of the
chick embryo was characterized by a high level of alkaline
phosphatase activity. Precocious phosphatase accumulation
induced by corticoid administration was accomplished by equally
precocious differentiation of the epithelial cells of duodenum.
The authors suggested that the functional differentiation of
the small intestine was normally controlled by secretions from
the cortical material of the embryo's own adrenal glands.

Salzgeber and Weber (1966) determined acid phosphatase and
cathepsin activity and total nitrogen in the homogenates of
chick embryo mesonephros during growth and regression. The
loss of total nitrogen in this period was as much as 62 per-
cent. Specific acid phosphatase activity increased appreciably
during regression.

e. Chick oviduct

Administration of exogenous estrogen diethyl-stilboestrol
dipropionate caused an increase in the alkaline phosphatase
activity of chick oviduct. Graded doses of estrogens caused
a relative increase in the alkaline phosphatase activity of
the oviduct which was significantly proportionate to the dose
of estrogen (Pande et al. , 1966).

f . Isozyme study with alkaline phosphatases

Characterization of alkaline phosphatases of the chick to
ascertain the origin of the serum enzymes and the enzymatic

24

relationship to calcium metabolism was made by Kuan et al.
(1966). Fractionation by ammonium sulfate precipitation and
ion exchange column chromatography were conducted. From
observations of the above experiments and determinations of
enzyme concentrations of the serum, bone, intestine, and liver
at various ages after hatching, the authors concluded that the
serum alkaline phosphatase isozymes originated from bone. The
chromatographic separation of proteins with enzyme activity
from bone behaved more like those of the serum than those of
the intestine and the liver.

Also see Maddaiah et al. (1964) — phytase, p. 25;

Hijmans and McCarty (1966) — invertase, p. 28;
Hinsch (1966) — ATPase, p. 31;

McDaniel and Chute (1961) — glutamic oxaloacetatic
transaminase, Class 2, p. 18.

(8) 5 '-Nucleotidase

Chicken muscle

The suitable conditions for the assay of 5 ' -nucleotidase
activity in chick skeletal muscle were described by Roffler
et al. (1966). They found that both calcified and normal
chick muscles were extremely low in enzyme activity.

Also see Hinsch (1966) —ATPase, p. 31.

(9) Phytase

Chicken intestine

Maddaiah et al. (1964), using calcium phytate and sodium phytate
as dietary phosphorus sources, measured the intestinal phytase
activity in chicks and mature hens. The measurement failed
to show a significant difference in the activity of this enzyme
due to the dietary treatments. It only indicated that the
phytase activity might have been due to a nonspecific phos-
phatase in the intestinal homogenates.

(10) Glucose -6 -phosphatase

Chick embryo

Hepatic glucose-6-phosphatase activity was found in developing
chick embryo almost simultaneously with the appearance of the
first hepatic cells. G-6-Pase and g-glucuronidase activity of
the embryo increased in direct proportion to liver weight.
Properties of embryonic liver G-6-Pase were similar to those
found in adult liver of other species. Studies with liver

25

slices indicated that the embryo liver was capable of glyco-
genolysis and glucose formation (Kilsheimer et al., 1960).

The effect of cortisone and hydrocortisone on glucose-6-
phosphatase activity in the chick embryo liver was studied by
Ogorodnikova (1965). Both hormones increased the activity of
the enzyme.

Also see Hellerstrom (1963) — alkaline and acid phosphatases,
P. 24;
Hinsch (1966) — ATPase, p. 31.

(11) Phosphoproteinphosphatase

a. Chick embryo

Phosphoproteinphosphatase from chicken embryos completely
split off alkaline-labile phosphorous from casein and the
Alkaline-labile phosphorous in brain phosphoprotein (Fedorova
and Komkova, 1964).

b. Chicken and pigeon muscle

Bargoni et al. (1963) found phosphoproteinphosphatase in the
soluble part of chicken and pigeon muscle cells.

(12) Phosphodiesterase

Chicken bone marrow

Bristow and Yamada (1965) found phosphodiesterase as well as
an acid phosphatase in the marrow of White Leghorn chickens.
Some properties and the intracellular distribution of these
two enzymes were described.

(13) Serine ethanolamine phosphate phosphodiesterase
Chicken kidney

Hagerman et al. (1965) purified serine ethanolamine phosphate
phosphodiesterase from chicken kidney. They believed that in
conjunction with kidney phosphomonoesterase, the new diesterase
accounted for the degradative metabolism of the diester.

(14) Deoxyribonuclease I and

(15) Deoxyribonuclease II
Chick embryo

The in vitro activity of deoxyribonuclease I and deoxyribo-
nuclease II was determined in the embryo and in the allantoic

26

fluid of the chick during the 6th to 16th day of development
within the egg. The respective increases in activities and
the successive changes in the ratios of activities of
deoxyribonuclease i/deoxyribonuclease II were reported
(Tempel and Zipf, 1966).

(16) Amylase

a. Chicken alimentary canal

A study on the distribution of enzymes in the alimentary
canal of the chicken showed the presence of amylase, invertase,
lactase, lipase, and proteolytic enzymes (Plimmer and Rosedale,
1922).

b. Chicken saliva

Amylase always was present in the saliva of mature hens,
lipase was present but relatively inactive, salivary protein-
ase was negligible or absent, and rennin was not present
(Leasure and Link, 1940).

c. Chicken bile and liver

An amylase which was secreted by the liver was found in chicken
bile. Its optimum hydrogen ion concentration was similar to
that of pancreatic amylase (Farner, 1943).

d. Chicken blood

Serum amylase activity in chickens was depressed by cholera
infections (Squibb et al., 1955).

e. Chick embryo

The growth pattern of the chick embryo was related to a,-amylase
activity in the developing pancreas. The steepest rise in
amylase activity coincided with the period of rapid growth and
increase in cell size which overlapped the time of hatching
(Kulka and Duksin, 1964). When stimulated by cholinergic
drugs, the pancreas taken from chick embryos would secrete
amylase in vitro. The pancreas evidently acquired the ability
to secrete digestive enzymes more than 10 days before the
beginning of the prominent biochemical and morphological
changes associated with the maturation of the gland (Kulka and
Yalovsky, 1966).

27

(17) Invertase

a. Chicken muscle

Invertase was present in the glycerine extracts of the chicken
gizzard muscle, the mucous membrane of the gizzard, and in the
thigh and breast muscle (Bernardi and Schwarz, 1932).

b. Chick embryo

Hydrocortisone induced the activity of alkaline phosphatase
and invertase but not lactic dehydrogenase in chick embryo
duodenum culture. The authors suggested that hormones might
act as specific inducers of genetic transcription of the syn-
thesis of a number of enzymes, and that some hormones might
also have an action on some step in protein synthesis beyond
the level of genetic transcription (Hijmans and McCarty, 1966).

Also see Plimmer and Rosedale (1922) — amylase, p. 27.

(18) Carboxypeptidase

Chicken blood and lymph

Erdos et al. (1964) described a carboxypeptidase which existed
in the blood sera of birds, mammals, amphibia, and other
animals; in lymph; in human urine and thrombocytes; but not
in erythrocytes. No significant enzyme activity was seen in
human serum.

(19) Pepsin

Chicken and pigeon gastric mucosa

Friedman (1939) reported that with respect to acid and pepsin,
the chief difference between mammals and birds was that in
birds (chicken and pigeon) both substances were produced by
the same type of cells, while in mammals each was produced by
one particular kind of cell — two morphologically different
elements of the mucosa of the stomach. The concentration of
pepsin was higher in the gastric juice of the chicken than in
that of the pigeon. Levchuk and Orekhovich (1963) purified
pepsinogen from chick gastric mucosa extract by means of ion
exchange column chromatography. Three pepsins were obtained.
Some of their hydrolytic properties were described.

(20) Trypsin and

28

(21) Chymo trypsin

a. Sea gull

Farner (1960) , in his review on the digestion and digestive
system of birds, stated that trypsin and intestinal entero-
kinase had been reported from the sea gull. He also stated
that protein digestion in birds, like so many other aspects
of avian digestion, needed extensive investigation.

b. Turkey and chicken

Purified turkey trypsin behaved in much the same manner as
bovine trypsin, both toward substrates and inhibitors.
Chicken chymotrypsin, however, exhibited some unusual behavior
toward natural and synthetic substrates as well as some
naturally occurring protease inhibitors. The differences in
the reactivity between chymotrypsins from avian and mammalian
origins toward the ester and protein substrates tested proba-
bly were due to primary or secondary structural variations
that were expressions of natural mutations (Ryan and Clary,
1964). Chicken chymotrypsin and turkey trypsin were further
purified (Ryan, 1965). Turkey trypsin reacted with substrates
and inhibitors in the same manner as mammalian trypsins. The
enzyme was stabilized by calcium at low and neutral pH.

The amino acid analysis also indicated a similarity to mammalian
trypsins. Chicken chymotrypsin showed esterase activity twice
that of bovine a-chymotrypsin but only two-thirds of the pro-
tease activity. Calcium did not increase the esterase activity
but did stabilize the enzyme near pH 7. At a low pH, the
enzyme was more stable in the absence of calcium. Results
indicated that natural variations between chicken and mammalian
chymotrypsins occurred in both physical and enzymatic prop-
erties, whereas variations between turkey and mammalian
trypsins were confined almost entirely to physical properties
(Ryan et al. , 1965).

c. Turkey ovomucoid

Simlot and Feeney (1964) showed that turkey ovomucoid inhib-
ited both trypsin and chymotrypsin and that the enzymes
occurred at independent sites.

(22) Elastase

Chick sera

Walford and Schneider (1959) presented the percent of inhibi-
tion of elastase in chicken sera.

29

(23) Arginase

Bird liver and kidney

Hunter and Dauphinee (1924) showed that arginase was absent
in the livers of birds but present in their kidneys. Dounce
(1950), however, stated that arginase activity was zero in
chicken liver cell nuclei. Cohen and Brown (1960), in their
comment on ammonia metabolism and urea biosynthesis, stated
that the location of arginase in the avian kidney instead of
the liver might serve to minimize the breakdown of arginine.
Arginine is an essential amino acid for birds. The presence
of arginase in kidney serves to provide ornithione for the
detoxication of benzoic acid.

Brown (1966), using more precise techniques, further demon-
strated that arginine occurred in the liver of numerous birds.
The stoichiometry of the reaction was demonstrated, and the
urea formed was derived from the amidine group of arginine.
A relatively high level of arginase activity was found in the
liver of some birds — kingfishers, herons, and gulls. The
lowest activity occurred in the liver of chickens and pigeons.

(24) Inorganic pyrophosphatase

Pigeon pancreas

Pynes and Younathan (1964) reported that the supernatant
fraction of the pigeon pancreas contained one of the highest
inorganic pyrophosphatase activities reported for tissues of
higher animals. The existence of this enzyme in the pancreas
had not been reported previously.

(25) ATPase

a. Pigeon muscle

Kitiyakara and Harman (1953) centrifugally fractionated pigeon
breast muscle and analyzed the distribution of enzyme activity
among the different particles. Magnesium-activated ATPase
(adenosinetriphosphatase) and myokinase were found to be
located predominantly in the cy tochondria. The myofibrillar
nuclear components were the sites of calcium-activated ATPase
and adenylic acid deaminase. Chappell and Perry (1953) showed
that pigeon-breast mitochondria contained an ATPase which was
activated to a greater extent by Mg++ than by Ca++.

b. Chicken muscle

Tonzetich and Kare (1960) differentiated adenosinetriphospha-
tase preparations obtained from two muscle sources of hen

30

muscle on the basis of their solubility. An active ATPase
was extracted with water from the red muscle, but not from
the white. Active preparations from both muscle sources
were soluble in dilute KC1. The KCl extracts of the water-
insoluble white-muscle sediment yielded ATPase activity
comparable to the direct KCl extracts. Dialysis of the enzyme
preparations failed to remove any factor necessary for ATPase
activity.

c. Chick embryo

Klein (1963) studied the ATPase of isolated mitochondria and
microsomes from embryonic chick ventricles at different stages
of development. He demonstrated a typical myocardial cation
transfer ATPase in the microsomal fraction.

d. Pigeon brain

The microsomal ATPase prepared from pigeon brain was activated
by Mg but little by Na and K. Twenty micromolar Cu achieved
maximum inhibition, reducing enzyme activity by approximately
25 percent. It was found that there were at least three com-
ponents in the pigeon's brain microsomal ATPase (Peters et al.,
1966). Peters and Walsh (1966), in their study of the toxic
action of copper using pigeon and rat brains, found that the
highest affinity for copper was in an ATPase-rich microsomal
fraction.

e. Chick esophagus and trachea

Tissues of chick embryos (17 to 45 days) and chicks (1 day to
3 weeks of age) were frozen and sectioned. Adenosinetriphos-
phatase, 5' -nucleotidase, acid phosphatase, nonspecific
esterase, nonspecific glycerophosphatase, nucleotide-diphos-
phatase, and glucose-6-phosphatase were localized in these
tissues (Hinsch, 1966).

f . Herring gull salt gland

Bonting et al. (1964) found Na-K-activated ATPase to a high
degree in the salt gland of the herring gull. The properties
of the enzymes were described.

Also see Lee (1963) — succinate dehydrogenase, Class 1, p. 12;
Hellerstrom (1963) — alkaline and acid phosphatases,
p. 24.

(26) Apryase

Chicken blood

Frank et al. (1950) studied apryases derived from chicken
blood. The enzyme activities were high in chicken red blood

31

cells and were associated with nuclei. The pH-activity curve
differed markedly among species. Magnesium ions stimulated
the activity of the chicken blood plasma uniformly throughout
the pH curve.

|

(27) Deoxycytidylate deaminase

Chick embryo

Maley and Maley (1963, 1964a) reported the purification and
properties of deoxycytidylate deaminase recovered from 6-day-
old chick embryo extract. The activity of the enzyme depended
upon the presence of deoxycytidine triphosphate and magnesium
ions. The authors suggested that thymidylate biosynthesis
from deoxycytidylate was regulated by end products associated
with its metabolic pathway, dCTP and dTTP. Later (1964b) they
showed that the activation effect by dCTP was associated with
an alteration in the structure of the enzyme.

(28) Fluoroacetanilide amidohydrolase

Chick liver

Nakamura et al. (1966b) extracted fluoroacetanilide amido-
hydrolase from the mouse and chick livers and compared their
basal properties. Enzymes from both sources were inhibited
by reagents that react with sulfhydryl group and chelate
with metallic ions, and their properties were mutually coin-
cidental. Nakamura et al. (1966a) studied the chick enzyme
further. It hydrolyzed monohaloacetanilides, iodoacetanilide
and chloroacetanilide, beside f luoroacetanilides, but it had
no activity with acetanilides. It was a metal enzyme con-
taining a tightly bound metal ion. The activity of the enzyme
was increased by Mn++, Mg++, CO++, B8++, and Ca"*-1" and was
decreased by Zn"*-1" and Ni"*"*".

(29) Adenosine deaminase

Chick

Chilson (1963) prepared chicken and calf adenosine deaminase.
He found subtle differences in the rate at which deoxyadenosine
was deaminated and more striking differences with the substrates
2-f luoroadenosine and 2 , 6-diaminopurine nucleoside. Also, the
temperature effect on the chicken's enzyme was different from
that on the calf's.

(30) g-Glucuronidase

See Kilsheimer et al. (1960) — glucose-6-phosphatase, p. 25-26.

32

(31) Adenylic acid deaminase

See Kitiyakara and Harman (1953) — ATPase, p. 30.

(32) Phosphomonoesterase

See Hagerman et al. (1965) — serine ethanolamine phosphate
phosphodiesterase, p. 26.

(33) Acetylcholinesterase

See Aprison et al. (1964) — hydroxytryptophan decarboxylase,
Class 4, p. 34.

(34) Uronolactonase

See Reithel (1967) — glucuronolactone reductase, Class 1,
p. 2.

(35) Formylase

See Knox and Eppenberger (1966) —tyrosine transaminase,
Class 2, p. 18.

(36) ATP pyrophosphatase

See Lee (1963) —succinate dehydrogenase, Class 1, p. 12.

(37) Nucleotide-diphosphatase

See Hinsch (1966)— ATPase, Class 3, p. 31.

(38) Fructose-diphosphatase

See Krebs (1964) — pyruvate carboxylase, Class 6, p. 37.

(39) Lactonase

See Goodridge and Ball (1966) —acetyl CoA carboxylase,
Class 6, p. 37-38.

4. Lyases

Enzymes of this group catalyze the removal of groups from substrates
without hydrolysis. They leave double bonds or conversely, add
groups to double bonds.

33

(1) 5 -Hydroxy tryptophan decarboxylase

Pigeon brain

Activities of 5-hydroxytryptophan decarboxylase, choline
acetylase, acetylcholinesterase, and monoamine oxidase were
determined in the telencephalon, diencephalon, plus optic
lobes, cerebellum, and ponsmedulla oblongata of young adult
pigeons. A comparative study of the enzymes was conducted
(Aprison et al., 1964).

(2) Aldolase

a. Chicken plasma

Cornelius et al. (1959), noticed a pronounced elevation of
plasma aldolase activity occurring in all birds exhibiting
dystrophy. The glutamic-oxaloacetic transaminase activity
also increased.

b. Pigeon muscle

George and Talesara (1962b) demonstrated histochemically the
presence of a high concentration of aldolase in the broad,
white glycogen-loaded fibers.

Also see McFarland et al. (1965) —lactate dehydrogenase, Class 1,
p. 5-6;
Rinaudo (1962) — phosphohexoisomerase, Class 5, p. 36,

(3) Citrate cleavage enzyme

a. Chicken blood

The oxygen consumption of the avian erythrocyte involved the
mechanism of the tricarboxylic acid cycle; all tricarboxylic
acid cycle enzymes were present except a specific oxalacetic
decarboxylase (Rubinstein and Denstedt, 1953).

b. Chicken liver

Srere and Bhaduri (1964) reported the purification and prop-
erties of the citrate cleavage enzyme from chicken liver.

Also see Goodridge and Ball (1966) —acetyl CoA carboxylase,
Class 6, p. 37-38.

(4) Carbonic anhydrase

Chicken oviduct

Common (1941) suggested that the carbonic anhydrase activity
of the uterine epithelium of hen was higher than that of the

34

remaining oviductal tissues, and that this activity might play
a part in shell secretion. Gutowska and Mitchell (1945) re-
ported that carbonic anhydrase seemed to be one of the active
factors in the enzymatic system of calcification of the egg-
shell. It liberated the anion, but not the cation, needed in
this process. The same hens could be made to lay either strong-
shelled, or soft-shelled eggs, or rough-shelled eggs by
inhibiting the activity of carbonic anhydrase by subcutaneous
sulfanilamide injection. The authors proposed that this enzyme
acted in the shell gland as a catalyst for decomposition of
carbonic acid, which might form bicarbonate ions from the blood;
these carbonate ions then were utilized in calcium carbonate
deposition. Robinson and King (1963) demonstrated histologically
that carbonic anhydrase was concentrated at the mammillae of
shell membrane.

Also see Bonting et al. (1964) — ATPase , Class 3, p. 31.

(5) D-Serine dehydratase

Chicken kidney

Grillo et al. (1965) partially purified D-serine dehydratase
from chicken kidney. Some general properties and activators
were reported.

(6) Cysteine synthetase

Chicken embryo (vitellin sack and liver)

Sentenac and Fromageot (1964) reported the presence of serine-
hydrolyase in the vitellin sack and the liver of chicken embryo.
This enzyme catalyzed the synthesis of cysteine from serine
and mineral sulfide.

(7) Phosphopyruvate dehydratase

Chicken embryo

Rinaudo (1962) studied the glycogenesis in the liver of chicken
embryo. Phosphopyruvate dehydratase, glyceraldehyde-3-phosphate
dehydrogenase, aldolase, and phosphohexoisomerase were assayed.
These enzymes behave differently from one another during the
6th to 20th day observed during the incubation, but they all
had one or two maxima of activities on or between the 12th and
the 10th days. After the 20th day all the activities dropped.
The author believed that his experimental data suggested the
reversal of anaerobic glycolysis as the pathway for the
glycogenesis from pyruvate in the liver of chicken embryo.

(8) 5-Aminoimidazole ribotide carboxylase

35

(9) 5-Amino-4-imidazole-N-succinocarboxamide ribotide cleavage
enzyme and

(10) Inosinicase

Chicken and pigeon liver

The above are the lyases among the enzymes of purine nucleotide
synthesis as given in the review by Flaks and Lukens (1963).

(11) Fumarase and

( 12 ) Aconitase

See Rubinstein and Denstedt (1953) — citrate cleavage enzyme,
Class 4, p. 34.

(13) Enolase

See Rinaudo (1962) — phosphohexoisomerase , Class 5, p. 36.

5. Isomerases

The enzymes in this group catalyze various types of isomerization.
They can be subdivided into racemases, epimerases, cis-trans
isomerases, intramolecular ketol isomerases, and intramolecular
transferases.

(1) Phosphohexoisomerase

Chick embryo

Rinaudo (1962) reported the activity of phosphohexoisomerase
in the chicken embryo liver.

( 2 ) Methylmalonyl-CoA mutase

Chick liver

A vitamin B]^ deficient liver evidenced lower methylmalonyl-
CoA mutase within 3 weeks in chicks. Isotopic tracings showed
that the hydrogen atom transfer that occurred during the mutase
reaction originated in the methyl group of methylmalonyl CoA
(Erfle et al. , 1964).

(3) Pho sphope n toe pime rase and

(4) Phosphor ibo isomer a se

See Goodridge and Ball (1966) — acetyl CoA carboxylase, Class 6,
p. 37-38.

36

6. Ligases

This group of enzymes catalyze the linking together of two molecules
with the breakdown of a pyrophosphate bond in ATP or a similar
triphosphate.

(1) Pyruvate carboxylase

a. Pigeon liver and kidney

Krebs (1964) in his 1963 Croonian lecture on "Gluconeogenesis"
mentioned that experiments on pigeon liver homogenates and kidney
cortex slices indicated that feedback control occurred in the
stage of pyruvate carboxylase and fructose-diphospha tase.

b. Chicken liver

Keech and Utter (1963) and Utter and Keech (1963) showed that
intracellularly pyruvate carboxylases appeared to be associated
with the particulate matter of avian liver cells. The
characteristics of the enzyme were described. The enzymatic
reaction did not appear to be related to the CC^-fixing
reaction. Later work showed that this enzyme catalyzed an
ATP- and acetyl CoA-dependent fixation of CO2 with pyruvate to
form oxaloacetate. The enzyme contained biotin (Scrutton and
Utter, 1964). The method which permitted the preparation of
relatively large quantities of pyruvate carboxylase from
chicken liver mitochondria was described. Some physical and
chemical properties of the highly purified enzyme were also
described (Scrutton and Utter, 1965).

(2) Acetyl CoA carboxylase

a. Chicken liver

Numa et al. (1966) prepared acetyl CoA carboxylase from chicken
liver. A correlation between activation of the carboxylase and
an increase in its sedimentation coefficient was shown.

b. Pigeon adipose tissue

Goodridge and Ball (1966) found low activities of acetyl CoA
carboxylase, citrate cleavage enzyme, malic enzyme, and hexose
monophosphate shunt dehydrogenases in the homogenates of pigeon
adipose tissue, but very high activities of those enzymes,
except for shunt dehydrogenases, in the homogenates of pigeon
liver, about 4 to 20 times higher than those in rat liver.
They also found that the marked effect of starvation and
refeeding on the enzyme activities of liver and adipose tissues
of rats was not present in those of pigeon. Since these enzymes
were involved in lipogenesis, the authors believed that pigeons
adipose tissue might be much less active in d_e novo lipogenesis

37

than rat adipose tissue and might serve largely as a depository
for fat synthesized elsewhere. They suggested that liver might
be the chief site of fatty acid synthesis in pigeons. Waite and
Wakil (1966) showed by using the criterion of the degree of
binding of the biotin derivatives to avidin that these compounds
had an intact ureido ring, thus eliminating the diamine deriva-
tive as an intermediate in the carboxylation.

(3) Glutamine synthetase

Pigeon liver

Sasaoka et al. (1964) discussed the possible presence of gluta-
mine synthetase in pigeon liver because of its ability to
synthesize theanine from glutamic acid and ethylamine in the
presence of ATP.

(4) Glycinamide ribotide kinosynthase

(5) Formylglycinamidine ribotide kinocyclodehydrase and

(6) 5-Amino-4-imidazole-N-succinocarboxamide ribotide kinosynthase

Chicken and pigeon liver

The above were the ligases among the enzymes of purine nucleotide
synthesis as given in the review by Flaks and Lukens (1963).

(7) Succinyl CoA synthetase

See Rubinstein and Denstedt (1953) —citrate cleavage enzyme,
Class 4, p. 34.

II. Miscellaneous Enzymatic Reactions

This section reports those works on enzymatic activities in which the
authors did not specify the enzymes or enzyme systems. The subject matter
is arranged chronologically according to the date of publication.

Cyclophorase

The skeletal bird muscles which were used only sporadically generally
had relatively low cyclophorase activity and a low mitochondrial count,
whereas muscles which had to function either continuously or for pro-
longed periods have a high cyclophorase activity and an abundance of
mitochondrial units (Paul and Sperling, 1952).

Dipeptidase

The nucleated erythrocytes of birds had a higher dipeptidase content
than that of the anucleated erythrocytes of mammals (Salvidio and Urbani,
1954). 3g

Carbohydrate metabolism

Chicken liver differed from rat liver in three things: 1. sucrose
enhanced the oxygen uptake of chicken liver slices incubated in a
saline buffered with phosphate but not those of rat liver, 2. sorbital
did not support chicken liver respiration as it did rat liver, and
3. the fructose metabolism end products were different. The chicken
end product was 1-a-glycerophosphate while the rat end products were
dihydroxyacetone phosphate and glyceraldehyde (Heald, 1962). Slices of
chicken liver metabolized fructose with an increased oxygen uptake, in
contrast with mammalian liver where fructose metabolism did not increase
the oxygen uptake. The RQ (respiratory quotient) was 1.1. Slices of
pigeon liver also showed an increased oxygen uptake, but RQ not greater
than 1.0 (Heald, 1963).

Esterases

Starch-gel electrophoresis was used in the study of the tissue-
and species-specificity of esterases. Chicken esterases exhibited
entirely different zymogram patterns from those of other species and
were highly reproducible (Paul and Fottrell, 1961). In agar electro-
phoresis the esterase activity of duck serum was found to localize mainly
in the region of a-globulin (Kaminski and Gajos, 1963). The esterases
activity was further characterized by Kaminski (1964). The isozyme
patterns of chicken pancreas were compared with those prepared from serum
and liver. The esterase patterns were found to be organ specific, but
the electrophoretic patterns for the splenic pancreatic lobe and the
ventral lobe showed no differences. A marked esterase reaction along
the capillaries of the A cell islets was noted (Hellman and Beckman,
1963).

Ribonucleic acid polymerase

In the study of the effect of estradiol injections upon chicken
liver nuclei ribonucleic acid polymerase, Weill et al. (1963) found
that after injection with estradiol, no change in composition of the
synthesized RNA could be detected through comparison of carbon-14
adenosine triphosphate and carbon-14 CTP incorporations. In addition,
the ratio of these incorporations was different from the CMP to AMP
ratio in chicken liver deoxyribonucleic acid.

Metalloporphyrin- forming enzymes

Metalloporphyrin-forming enzymes were found in the extracts of
avian erythrocytes. While liver mitochondrial extract readily
catalyzes the incorporation of Fe^+ and Co^+ into protoporphyrin, the
extract of avian erythrocyte only catalyzes the incorporation of Fe^+
(Johnson and Jones, 1964).

39

Fatty acid synthetase

The independent binding of the acetate and malonate moieties of
acetyl and malonyl coenzyme A to a pigeon liver fatty acid synthetase
through thioester bonds was demonstrated. The evidence supported a
scheme of fatty acid synthesis which involves the stepwise addition of
two carbon units from malonyl -enzyme to acyl -enzyme with the formation
of 0-ketoacyl-enzyme (Brodie et al., 1964).

Acylating enzymes

Pigeon pancreas microsomal and mitochondrial fractions catalyzed the
formation of phosphatidylinosital from fatty acid thioesters of coenzyme A
and lysophosphatidylinositalo An easy method was described (Keenan and
Hokin, 1964).

Sacharide polymerase

A cell -free enzyme preparation was obtained from chick embryo epiph-
yses. It catalyzed the synthesis of a high molecular weight polysacharide
from uridine 5 ' -diphosphate-N-acetyl-D-galactosamine, or UDP-N-acetyl-D-
glucosamine and UDP-D-glucuronic acid. When the particulate enzyme was
washed twice, the UDP-GalNAc pyrophosphorylase and UDP-GlcNAc-4 ' -epimerase
were removed, while the polymerase activity was retained (Perlman et al . ,
1964).

Anserine-hydrolyzing enzyme

The anserine-hydrolyzing activity in serum and acetone fraction pre-
pared from pectoral muscles, leg muscles, livers, and kidneys of Japanese
quail and variously-aged chickens was investigated and the activity was
recognized in the kidney. The enzyme also hydrolyzed carnosine and
orphidine (Ishikawa et al. , 1964).

Benzoylornithines synthesizing enzymes

The synthesis of monobenzoylornithine and ornithuric acid in the
presence of benzoic acid, ornithine, adenosine triphosphate, and coenzyme
A was catalyzed by an enzyme system prepared from chicken kidney. Pre-
liminary fractionation studies suggested the involvement of at least three
enzymes in the over-all reaction (Marshall and Koeppe, 1964).

Glycogen synthetase

See Fridland and Nigam (1965) — phosphorylase, Class 2, p. 16.

Glycyl-soluble-RNA synthetase

A glycyl-soluble-RNA synthetase was prepared from chick embryo. It
catalyzed a glycine-dependent 32PPi--ATP exchange but not 32Pi--ATP ex-
change, and the formation of glycyl-soluble-RNA. The enzyme required
either Mg2+ or Mn2+ for activity; Mg2+ was more effective at high concen-
tration and Mn2+ at low concentration. Ca2+ antagonized the action of

Mg2+ (Bublitz, 1966).

40

Amylase

Omission of Ca^+ from the incubation medium completely prevented the
acetylcholine-stimulated release of amylase into the incubation medium.
The omission caused some increase in the incorporation of phorphorus-32
into the lipides (Hokin, 1966).

Long-chain acyl desaturase

Analysis of the fatty acid formed in the in_ vitro synthesis by micro-
somal and soluble enzymes of pigeon liver showed that the long-chain acyl
desaturase activity was eliminated by 72 hours of starvation (Butterworth
et al., 1966).

Dehydrogenases and hydrolases

Electrophoretric zymograms were prepared from the aqueous extracts of
breast muscle, leg muscle, cardiac muscle, liver, kidney, and spleen of
adult cockerels. A partial characterization of the tissue esterases was
carried out with various substrates and specific inhibitors. The 18 bands
of esterases were aliesterases of two different types. Tissue-specific
enzymatic patterns were obtained for esterases, alkaline phosphatase, acid
phosphatase, lactate dehydrogenase, malate dehydrogenase, and isocitrate
dehydrogenase. Glutamate dehydrogenase and glucose-6-phosphate dehydro-
genase activity were detected only in the liver and kidney (Ecobichon, 1966)

Cathepsin

See Salzgeber and Weber (1966) — phosphatase, Class 3, p. 24.

DISCUSSION

This survey has shown that most of the work done on enzymes in birds
was incidental to other objectives. Little research has been devoted
exclusively to the study of enzymes or enzyme systems in birds, with the
objective of obtaining a better understanding of bird metabolism. Birds
used in research were mostly domestic — chickens in many cases, and some-
times pigeons, either domestic or wild. The tissues studied most were
muscles and liver.

Of the 120 enzymes surveyed, oxidoreductases and hydrolases are the
two classes investigated most thoroughly. All the enzymes in the pentose
shunt and citric acid cycle have been reported in the literature.
a-Ketoglutarate dehydrogenase, however, is not listed in APPENDIX B
because it is actually an enzyme complex. Of the enzymes related to
nitrogen metabolism in birds, references could be found only on those
associated with nucleotide metabolism. Enzymes involved in nitrogen
metabolism are of much interest in bird control because instances have been

41

found of the diversity in nitrogen metabolism among species. These dif-
ferences, when discovered, might be exploited for the purpose of selective
toxication. It is regrettable, therefore, that no useful data on these
enzymes in birds could be found.

Some comparison of the degree of intensity of enzymatic activities
among species have been found, but the works have been few. During recent
years, comparative enzymology has been exploited as a means of taxonomy
(e.g., Wilson and Kaplan, 1964). Such attempts with birds have been made
especially with lactate and malate dehydrogenases. Study in this area is
still quite limited, however.

Since birds are said to differ characteristically from mammals in
excreting uric acid instead of urea as a waste product, a fundamental
difference in the nitrogen metabolism including the urea cycle must exist
between the two classes. Indeed, studies had been made to locate and
quantify arginase, but enzymes in the urea cycle have not been investigated
thoroughly. A study of bird metabolism and the enzymes concerned would
most likely yield enlightening results.

This survey also revealed that although comparative studies on the
enzyme biochemistry of birds are carried out sporadically, the number of
species investigated has, indeed, been small. As a whole, the field of
the comparative enzyme biochemistry of birds remains a virgin land. It
is understandable, therefore, that no such study has ever been conducted
on the species of birds that we are most interested in controlling. Such
comparative data are very important to our development of selective toxi-
cants for nuisance birds.

While Williams (1958) and Florkin (1960) reported on the biochemical
individuality and the unity and diversity in biochemistry, and Dixon and
Webb (1958), Dittmer (1961), and George and Berger (1966) tabulated and
mentioned some enzymes found ,in birds, it has remained for this survey to
collect and compile the available data on enzymes in birds covering a
wider area, giving more detail about individual enzymes concerned, in order
to facilitate intra- and inter-species comparison.

SUMMARY

Enzymes which appear in the literature concerning research with birds
have been surveyed and reported in the six classes that were recommended
in 1964 by the International Union of Biochemistry. The pertinent bio-
chemistry is described for each enzyme. A total of 120 enzymes was
surveyed.

The major comparative aspects of enzymes are as follows:

1. Avian livers have the highest known purine synthetic enzymes of
any animal tissue studied.

42

2. Malate oxidase, succinoxidase and malate dehydrogenase are more
active in bird erythrocytes than in those of higher vertebrates.

3. Birds have the highest levels of D-phosphoglycerate dehydro-
genase.

4. Chicken livers differ qualitatively and quantitatively from
rat livers in metabolizing fructose. Oxygen uptake did not increase in
the mammalian metabolism of fructose.

5. Arginase, the enzyme which catalyzes the change of L-arginine
to L-ornithine and urea, occurs in bird kidneys instead of livers as it
does in other animals.

6. Contrary to mammals, ornithine transcarbamylase is either low
or not detectable in birds by present methods of assay.

7. Chicken heart mitochondrial malate dehydrogenase is devoid of
tryptophan as is that of pig and horse.

8. The red-whiskered bulbul (Aves) , like the guinea pig, does not
have the microsomal enzymes necessary in the synthesis of ascorbic acid.

9. Hummingbirds and swifts heart extracts have a different zymo-
gram pattern of the "supernatant" form of malate dehydrogenase from that
of chicken enzyme on gel electrophoresis; all families of Charadriiformes
have an enzyme that moves more slowly than that of chicken enzyme.

10. Mammalian oxidase oxidizes quinine more rapidly than quinidine,
but the reverse is true for the enzyme in birds.

11. Pigeon liver has been found to contain no xanthine oxidase, and
thus it cannot oxidize hypoxanthine to uric acid.

43

LITERATURE CITED

Allison, W. S.

1964. The comparative enzymology of triosephosphate dehydrogenase.
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Allison, W. S., and N. 0. Kaplan.

1964. Effect of tetrathionate on the stability and immunological
properties of muscle triosephosphate dehydrogenases. Biochemistry,
vol. 3, p. 1792-1800.

Aprison, M. H. , R. Takahashi, and T. L. Folkerth.

1964. Biochemistry of the avian central nervous system: I. The
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acetylase-acetylcholinesterase systems in several discrete areas
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Augustinsson, K. -B.

1959a. Electrophoresis studies on blood plasma esterases. II. Avian,
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1959b. Electrophoresis studies on blood plasma esterases. III. Con-
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Bargoni, N. K. , T. Fossa, and A. Sisini.

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Beenakkers, A. M. Th. , and M. Klingenberg.

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Bell, D. J.

1960. Tissue components of the domestic fowl. 4. Plasma-alkaline-
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1961. Plasma acid phosphatase activity and bone dystrophies in the
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Berheim, F.

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Bernardi, A., and M. A. Schwarz.

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44

Bokdawala, F. D. , and J. C. George.

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61

ADP

ADH

AICAR

AMP

ATP

ATPase

CoA

DBA4"*"

dCTP

DNA

DPN DPNH

dTTP

FGAM

FGAR

GalNAc

GAR

GlcNAc

G6P

G6Pase

LDH

MDH

NAD NADH

NADP NADPH

APPENDIX A
Abbreviations in this report

adenos inediphosphate

alcohol dehydrogenase

5-amino-4-imidazole-carboxamide ribotide

adenos inemonophosphate

adenosine triphosphate

adenos inetriphosphatase

coenzyme A

10,10' -dimethyl -9, 9 ' -biacridylium ion

deoxycysteinetriphosphate

deoxyribonucleic acid

diphosphonucleotide, oxidized and reduced forms
(same as NAD and NADH)

deoxy thymine triphosphate

formylglycinamidine

formylglycinamide ribotide

N-acetyl-D-galactosamine

glycinamide ribotide

N-acetyl-D -glucosamine

glucose -6 -phosphate

glucose-6 -phosphatase

lactate dehydrogenase

malate dehydrogenase

nicotinamide adenine dinucleotide (coenzyme I, also
called DPN) oxidized and reduced forms

nicotinamide adenine dinucleotide phosphate (coenzyme II,
also called TPN) oxidized and reduced forms

62

PP pyrophophate

PRA phosphoriboxylamide

PRPP phosphoribose pyrophosphate

RQ respiratory quotient, defined as the ratio of the
volume of O2 absorbed over some time period

THFA tetrahydrofolate

TPD triosephosphate dehydrogenase

UDP uridinediphosphate

UDPG uridinediphosphoglucose

IMP uridinemonophosphate

63

APPENDIX B

Enzymes cited in this report*
(with recommended nomenclatures and reactions)

TRIVIAL NAME

Class 1. Oxidoreductases
SYSTEMATIC NAME

REACTION

Alcohol dehydrogenase Alcohol :NAD oxidoreductase

Alcohol + NAD = aldehyde or
ketone + reduced NAD

Catalase

Cystine reductase

Cytochrome c reductase

Cytochrome oxidase

Dihydrofolate
dehydrogenase

Folic acid reductase

Hydrogen-peroxide : hydrogen-
peroxide oxidoreductase

Reduced-NAD : L-cys tine
oxidoreductase

Reduced-NAD : (acceptor)
oxidoreductase

Ferrocytochrome c: oxygen
oxidoreductase

7 , 8-Dihydrof olate : NADP
oxidoreductase

5,6,7, 8 -Tetrahydrof olate
NADP oxidoreductase

H2O2 + H2O2 = 02 + 2 H20

Reduced NAD + L-cystine =
NAD + 2 L-cysteine

Reduced NAD + acceptor =
NAD + reduced acceptor

4 Ferrocytochrome c + O2 =
4 f erricytochrome £ + 2 H2O

7, 8 -Dihydrofolate + NADP =
folate + reduced NADP

5,6, 7, 8 -Tetrahydrof olate +
NADP = 7,8-dihydrofolate +
reduced NADP

Glucose dehydrogenases

Glucose -6 -phosphate
dehydrogenase

Glucuronolactone
reductase

B-D-Glucose:NAD(P)
oxidoreductase

D-Glucose-6-phosphate :NADP
oxidoreductase

L-Gulono-y-lactone :NADP
oxidoreductase

P-D-Glucose + NAD(P) =
D-glucono-8-lactone +
reduced NAD(P)

D-Glucose 6-phosphate + NADP =
D-glucono-6-lactone 6-phosphate
+ reduced NADP

L-Gulono-Y-lactone + NADP =
D-glucurono-Y-lactone +
reduced NADP

Glutamate dehydrogenase

L-Glutamate:NAD
oxidoreductase (deaminating)

L-Glutamate + H2O + NAD
2-oxoglutarate + NH3 +
reduced NAD

*The systematic names and reactions are given according to the 1964 recommendations,
Report of Commission on Enzymes, International Union of Biochemistry. The trivial
names are those of the authors.

64

TRIVIAL NAME

SYSTEMATIC NAME

REACTION

Glutathione
dehydrogenase

Glutathione :dehydro-
ascorbate oxidoreductase

2 Glutathione + dehydro-
ascorbate = oxidized
glutathione + ascorbate

Glutathione peroxidase

Glutathione : hydrogen-
peroxide oxidoreductase

2 Glutathione + H2C7 =
oxidized glutathione + 2 H2O

Glycerol -3 -phosphate
dehydrogenase

L-Glycerol- 3 -phosphate: NAD
oxidoreductase

L-Glycerol 3-phosphate + NAD
= dihydroxyacetone phosphate
+ reduced NAD

Glycerophosphate
dehydrogenase

L-Glycerol-3-phosphate :
(acceptor) oxidoreductase

L-Glycerol 3-phosphate +
acceptor = dihydroxyacetone
phosphate + reduced acceptor

3-Hydroxybutyrate
dehydrogenase

D-3-Hydroxybutyrate : NAD
oxidoreductase

D-3-Hydroxybutyrate + NAD =
acetoacetate + reduced NAD

Isocitrate
dehydrogenase

threo-Dg -Isocitrate: NAD
oxidoreductase

threo-Dg -Isocitrate + NAD
2-oxoglutarate + CO2 +
reduced NAD

Lactate dehydrogenase L-Lactate:NAD oxidoreductase

L-Lactate + NAD = pyruvate +
reduced NAD

Malate dehydrogenase

L-Malate:NAD oxidoreductase

L-Malate + NAD = oxaloacetate
+ reduced NAD

Malate oxidase

L-Malate : oxygen

L-Malate +02= oxaloacetate
+ (?)

Monoamine oxidase

Monoamine : oxygen
oxidoreductase (deaminating)

A monoamine + H2O +02=
an aldehyde + NH3 + H2O2

Oestradiol
17-3-dehydrogenase

Oestradiol :NAD

17 -3 -oxidoreductase

Oestradiol + NAD = oestrone
+ reduced NAD

Phosphogluconate
dehydrogenase

6-Phospho-D-gluconate:
NAD(P) oxidoreductase
(Decarboxylating)

6-Phospho-D-gluconate + NAD(P)
= 6-phospho-2-keto-D-
gluconate + reduced NAD(P)

Succinate dehydrogenase

Succinate: (acceptor)
oxidoreductase

Succinate + acceptor =
fumarate + reduced acceptor

Trios ephosphate
dehydrogenase

D-Glyceraldehyde-3-
phosphate : NADP
oxidoreductase

D-Glyceraldehyde 3-phosphate
+ NADP + H20 = 3-phospho-D-
glycerate + reduced NADP

Tryptophan pyrrolase

L-Tryptophan: oxygen
oxidoreductase

L-Tryptophan +02=
L-formylkynurenine

Uricase

Urate: oxygen oxidoreductase

Urate +02= unidentified
products

65

TRIVIAL NAME

SYSTEMATIC NAME

REACTION

Xanthine oxidase

Xanthine : oxygen
oxidoreductase

Xanthine + H2O +02= urate +
H202

5 -Amino -4 -imidazole -
carboxamide ribotide

Class 2. Transferases

L-arginine : glycine
amidino- transferase

AICAR + 10-formyl-THFA
Formyl-AICAR + THFA

Carnitine

acetyl transferase

Catechol

methyl transferase

Choline acetylase

Creatine kinase

Formylglycinamide
ribotide amido-
transferase

Acetyl -CoA: carnitine
0-acetyl transferase

S_-Adenosylmethionine :
catechol 0-methyl trans-
ferase

Acetyl -CoA: choline
0-acetyl transferase

ATP:creatine phospho-
transferase

Acetyl -CoA + carnitine =
CoA + O-acetylcarnitine

S_-Adenosylmethionine + catechol
= S_-adenosylhomocysteine +
guiacol

Acetyl -CoA + choline = CoA +
0 -acetylcholine

ATP + creatine = ADP + phos-
phocreatine

FGAR + L-glutamine + ATP +
H20 = FGAM + L-glutamic acid +
ADP + Pi

or-Glucan phosphorylase

Glutamic-oxaloacetic
transaminase

Glutamic -pyruvic
transaminase

Glycinamide ribotide
trans formyl as e

Hexokinase

Myokinase
NAD kinase

a-l,4-Glucan:orthophosphate
glucosyl transferase

L-Aspartate:2-oxoglutarate
aminotransferase

L-Alanine : 2 -oxoglutarate
aminotransferase

ATP:D-hexose 6-phospho-
transf erase

ATP: AMP phosphotransferase

ATP:NAD 2 '-phospho-
transferase

(a?-l,4-Glucosyl)n + orthophos-
phate = (a-l,4-glucosyl)n_i +
o/-D-glucose 1 -phosphate

L-Aspartate + 2-oxoglutarate =
oxaloacetate + L-glutamate

L-Alanine + 2-oxoglutarate =
pyruvate + L-glutamate

GAR + 5, 10 -methylene -THFA +
H20 = FGAR + THFA + H+

ATP + D-hexose = ADP + D-hexose
6-phosphate

ATP + AMP = ADP + ADP

ATP + NAD = ADP + NADP

Ornithine carbamoyl -
transferase

Carbamoyl phosphate : L-
ornithine carbamoyl-
transferase

Carbamoyl phosphate + L-orni-
thine = orthophosphate +
L-citrulline

66

TRIVIAL NAME

SYSTEMATIC NAME

REACTION

Phosphoglucomutase

Phosphoglycerate
mutase

a-D -Glucose -1 ,6 -diphos-
phate :o/-D-glucose-l-phos-
phate phosphotransferase

2 , 3-Diphospho-D-glycerate :

2-phospho-D-glycerate

phosphotransferase

or-D-glucose 1,6-diphosphate +

a-D-glucose 1 -phosphate =

oz-D-glucose 6-phosphate +

a-D-glucose 1,6-diphosphate

2, 3-Diphospho-D-glycerate +
2-phospho-D-glycerate =
3-phospho-D-glycerate +
2, 3-diphospho-D-glycerate

Phosphor ibosylpyro-
phosphate amido-
transferase

5 -Phosphor ibosylpyro-
phosphate amidotrans-
ferase

Ribosyl amine -5 -phosphate:
pyrophosphate phosphoribo-
syltransf erase (glutamate-
amidating)

P-Ribosylamine 5-phosphate +
pyrophosphate + L-glutamate =
L-glutamine + 5-phospho-or-D-
ribosylpyrophosphate + H2O

PRPP + L-glutamine + H20 =
PRA + L-glutamic acid + PP

Tyrosine -pyruvate
aminotransferase

L-Tyrosine : pyruvate
aminotransferase

L-Tyrosine + pyruvate =
p-hydroxyphenylpyruvate +
L-alanine

UDPglucose -glycogen
glucosyl transferase

UDPglucose : glycogen
Of-A -glucosyl transferase

UDPglucose + (glycogen)n
UDP + (glycogen)n+1

Acetylcholinesterase

Class 3. Hydrolases
Acetylcholine hydrolase

Acetylcholine + H2O = choline
+ acetate

Acid phosphatase

Orthophosphoric monoester
phosphohydrolase

An orthophosphoric monoester +
H2O = an alcohol + orthophos-
phate

Adenosine deaminase

Aliesterase

Adenosine aminohydrolase

Carboxylic-ester hydrolase

Adenosine + H2O = inosine +
NH3

A carboxylic ester + H2O = an
alcohol + a carboxylate

Alkaline phosphatase

Orthophosphoric monoester
phosphohydrolase

An orthophosphoric monoester +
HoO = an alcohol + orthophos-
phate

AMP deaminase
Of-Amylase

AMP aminohydrolase

or-l,4-Glucan 4-glucano-
hydrolase

AMP + H20

IMP + NH,

Hydrolyses o/-l ,4-glucan links
in polysaccharides containing
three or more o/-l,4-linked
D-glucose units

67

TRIVIAL NAME

SYSTEMATIC NAME

REACTION

Apyrase
Arginase

ATP diphosphohydrolase

L-Arginine amidinohydrolase

ATP + H20 = ADP + ortho-
phosphate

L-Arginine + H2O = L-
ornithine + urea

ATPase

ATPase

Carboxypeptidase A

ATP phosphohydrolase

ATP pyrophosphohydrolase

Pep t idyl -L-amino -acid
hydrolase

ATP + H20 = ADP + ortho-
phosphate

ATP + H20 = AMP + pyrophos-
phate

A peptidyl -L-amino acid +
H20 = a peptide + an L-amino
acid

Chymotrypsin A

Hydrolyses peptides, amides,
esters, etc., especially at
bonds involving the carboxyl
groups of aromatic L-amino
acids

Chymotrypsin B

Deoxyribonuclease I

Deoxyribonuclease II

Elastase

Deoxyribonucleate
oligonucleotidohydrolase

Deoxyribonucleate

3' -nucleotidohydrolase

Specificity similar to chymo-
trypsin A

DNA + (n-l)H20 = n oligodeoxy-
ribonucleotides + 5 '-monophos-
phates

Forms 3 ' -nucleotides and 3'-
monophosphates from DNA

Hydrolyses peptides, especially
at bonds adjacent to neutral
amino acid residues

Formylase

Glucose -6 -phosphatase

P- Glucuronidase

Inorganic pyrophos-
phatase

Invertase

Lactase

Aryl-formylamine amido-
hydrolase

D -Glucose -6 -phosphate
phosphohydrolase

g-D-Glucuronide
glucuronohydrolase

Pyrophosphate phospho-
hydrolase

8-D-Fructofuranoside
f ructohydrolase

B-D-Galactoside
galactohydrolase

N-Formyl-L-kynurenine + H20 =
formate + L-kynurenine

D-Glucose 6-phosphate + H2O =
D-glucose + orthophosphate

8-D-glucuronide + H2O = an
alcohol + D-glucuronate

Pyrophosphate + H20 = 2 ortho-
phosphate

A B-D-fructofuranoside + H2O =
an alcohol + D-fructose

A B-D-galactoside + H20 = an
alcohol + D-galactose

68

TRIVIAL NAME
Lipase

5 '-Nucleotidase

Pepsin

Pepsin B
Phosphodies terase

Phosphoprotein
phosphatase

Phytase

SYSTEMATIC NAME
Glycerol-ester hydrolase

5 '-Ribonucleotide phospho-
hydrolase

Orthophosphoric diester
phosphohydrolase

Phosphoprotein phospho-
hydrolase

meso-Inositol-hexaphosphate
phosphohydrolase

Pseudo-cholinesterase Acylcholine acyl -hydrolase

Trypsin

REACTION

A triglyceride + H2O = a
diglyceride + a fatty acid
ion

A 5' -ribonucleotide + HoO =
a ribonucleoside + ortho-
phosphate

Hydrolyses peptides, including
those with bonds adjacent to
aromatic or dicarboxylic L-
amino acid residues

Hydrolyses peptides; the
specificity is similar to
that of pepsin

A phosphoric diester + H2O =
a phosphoric monoester + an
alcohol

A phosphoprotein + n H2O = a
protein + n orthophosphate

meso- Inositol hexaphosphate +
6 HoO = meso-inositol +
6 orthophosphate

An acylcholine + H£0 = choline
+ an anion

Hydrolyses peptides, amides,
esters, etc., at bonds in-
volving the carboxyl groups
of L-arginine or L-lysine

Aconitate hydratase

Aldolase

Class A. Lyases

Citrate (isocitrate) hydro- Citrate = cis-aconitate + H2O
lyase

Ketose-1 -phosphate
aldehyde -lyase

5-Amino-4-imidazole-
N-succinocarboxamide
ribotide cleavage enzyme

5-Aminoimidazole
ribotide carboxylase

Carbonic anhydrase

Carbonate hydro-lyase
69

A ketose 1 -phosphate =
dihydroxyacetone phosphate +
an aldehyde

Succino-AICAR
fumaric acid

AICAR +

AIR + CO^

CAIR

H2C03(or H+ + HC03 )=
C02 + H20

TRIVIAL NAME

SYSTEMATIC NAME

REACTION

Citrate-cleavage enzyme

Cysteine synthetase

Enolase

Fumarate hydratase
Hydroxy tryptophan

Inosinicase

Oxaloacetate
decarboxylase

D-Serine dehydratase

ATP: citrate oxaloacetate-
lyase (CoA-acetylating and
ATP-dephosphorylating)

L-Serine hydro-lyase
(adding hydrogen sulphide)

2 -Phospho-D-gly cerate
hydro-lyase

L-Malate hydro-lyase

5 -Hydroxy-L- tryptophan
carboxy-lyase

Oxaloacetate carboxy-lyase

D-Serine hydro-lyase
(deaminating)

ATP + citrate + CoA = ADP +
orthophosphate + acetyl -CoA
+ oxaloacetate

L-Serine + H2S = L-cysteine
+ H20

2-Phospho-D-glycerate =
phospho-enolypyruvate + H„0

L-Malate = fumarate + H20

5-Hydroxy-L-tryptophan =
5 -hydroxy tryptamine + C02

Formyl-AICAR = IMP + H2O

Oxaloacetate = pyruvate + C02

D-Serine + H20 = pyruvate +
NH3 + H20

Methylmalonyl-CoA
mutase

Phosphohexoisomerase

Class 5. Isomerases

Methylmalonyl -CoA
CoA-carbonylmutase

D -Glucose -6 -phosphate
ketol-isomerase

Methylmalonyl -CoA = succinyl-
CoA

D-Glucose 6-phosphate =
D-fructose 6-phosphate

Class 6. Ligases

Acetyl -CoA carboxylase Acetyl -CoA: carbon-dioxide

ligase (ADP)

5 -Amino -4- imidazole -N-
succinocarboxamide
ribotide kinosynthase

Formylglycinamidine ribotide
kinocyclodehydrose

ATP + acetyl -CoA + CO2 + H20
= ADP + orthophosphate +
malonyl-CoA

CAIR + L-aspartic acid + ATP
= succino-AICAR + ADP + Pi

FGAM + ATP = AIR + ADP + Pi

Glutamine synthetase L-Glutamate: ammonia ligase ATP + L-glutamate + NH3

Glycinamide ribotide
kinosynthase

Pyruvate carboxylase

(ADP)

Pyruvate : carbon-dioxide
ligase (ADP)

ADP
+ orthophosphate + L-glutamine

Glycine + ATP + PRA = GAR +
ADP + Pi

ATP + pyruvate + C02 + H20 =
ADP + orthophosphate +
oxaloacetate

70

U.S. GOVERNMENT PRINTING OFFICE: 1971-437-570/129

As the Nation's principal conservation agency, the Department
of the Interior has basic responsibilities for water, fish, wildlife,
mineral, land, park, and recreational resources. Indian and Ter-
ritorial affairs are other major concerns of this department of
natural resources.

The Department works to assure the wisest choice in managing
all our resources so that each shall make its full contribution to
a better United States now and in the future.

UNITED STATES

DEPARTMENT OF THE INTERIOR

FISH AND WILDLIFE SERVICE

BUREAU OF SPORT FISHERIES AND WILDLIFE

WASHINGTON. D. C. 20240

POSTAGE AND FEES PAID

DEPARTMENT OF THE INTERIOR