Protein and Amino Acid Requirements of Mammals

158 Pages·1950·9.31 MB·English

PROTEIN AND AMINO ACID
REQUIREMENTS OF MAMMALS
Edited by
ANTHONY A. ALBANESE
Nutritional Research Laboratory, St. Luke's Hospital,
New York, New York
1950
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CONTRIBUTORS
ANTHONY A. ALBANESE, Nutritional Research Laboratory, St. Luke's
Hospital, New York, New York.
BACON F. CHOW, Department of Biochemistry, School of Hygiene and
Public Health, The Johns Hopkins University, Baltimore, Maryland.
DOUGLAS V. FROST, Abbott Laboratories, North Chicago, Illinois.
H. H. MITCHELL, Division of Animal Nutrition, University of Illinois,
Urbana, Illinois.
CURT C. PORTER, Merck Institute for Therapeutic Research, Rahway,
New Jersey.
ROBERT H. SILBER, Merck Institute for Therapeutic Research, Rahway,
New Jersey.
Poreword
The past decade has witnessed a dramatic increase in public and
scientific interest in nutrition. Because of the vital nature of protein in
animal and human economy, a greater share of scientific attention has
been directed to these food components than to any other moiety of the
diet.
One of the major problems, if not the most important problem in
nutrition, concerns the determination of the protein and amino acid
requirements of various mammals, and man in particular. The ap
proaches to this complex problem are numerous. It is the purpose of
the volume to describe the techniques currently employed in this field,
and to point out frankly their limitations. The reader will discover
that although much has been accomplished, much more needs to be
done before completely satisfactory values will be available on the
protein and amino acid needs of mammals during growth, adulthood
and senescence.
ANTHONY A. ALBANESE
New York, N. Y.
September, 1950
vii
Some Species and Age Differences
in Amino Acid Requirements
H. H. MITCHELL
Division of Animal Nutrition, University of Illinois, Urbana, Illinois
CONTENTS
I. Introduction 1
II. An Experimental Study 3
1. Experimental Methods 4
2. Experimental Results 4
3. Interpretations 5
4. A Test of the Amino Acid Adequacy of Whole Egg Proteins . .. 5
5. The Adult Rat Compared with Adult Man 7
III. Keratin Synthesis in Protein Nutrition 9
1. The Raiding of Protoplasmic Tissue for Methionine-Cystine . . .. 10
2. Wool Growth 11
3. Further Evidence and Recapitulation 12
IV. Composition of Tissue Proteins and Amino Acid Requirements for Growth 13
1. Similarities in the Amino Acid Content of Animal Tissue Proteins . 13
2. Biological Confirmation 15
3. Comparison with Growth Requirements 16
V. Amino Acid Requirements for Nitrogen Equilibrium in the Adult . .. 18
VI. A Theory of Protein Metabolism 20
1. Basic Principles 21
2. The Endogenous Protein Catabolism 22
3. Protein Stores in the Body 22
4. The Role of the Blood Plasma Proteins 23
5. Keratin Synthesis 23
6. The Exogenous Protein Catabolism Studied by the Isotope Technic . 24
VII. A Schematic Representation of Protein Metabolism 25
VIII. Summary 28
References 29
I. INTRODUCTION
The unique functions in the animal body served by the amino acids
resulting from protein digestion are all anabolic in character. They
relate to the replacement of essential tissue constituents that have been
degraded in catabolic reactions, or to the formation of new tissue con
stituents in growth. In the rapidly growing animal, the latter functions
dominate the body's requirements for amino acids. In the mature ani
mal, the replacement functions may dominate the amino acid require-
1
2 H. H. MITCHELL
ments, but the growth functions still persist, since some tissues continue
to grow throughout life (1). Among adult animals of different species,
the relative importance of the growth functions in determining amino
acid requirements will depend mainly on the rate of growth of the epi
dermal structures, such as hair, nails and claws.
The different types of anabolic reactions occurring in the growing
and in the adult organism with reference to the disposal of dietary amino
acids lead inevitably to the conclusion that the amino acid requirements
of infancy and adolescence, on the one hand, and of maturity, on the
other hand, are different either in the assortment of amino acids that are
required preformed in the diet, or in the relative amounts that are
needed, or both. Species differences in amino acid requirements during
growth would be expected not so much on the basis of differences in the
nature of the anabolic reactions, as on the basis of differences in the rate
of growth of new tissue in comparison with the weight of tissue to be
maintained, that is, the percentage rate of growth. The higher the per
centage growth rate, the greater the extent to which the growth of new
tissue dominates the total amino acid requirements. The rat, growing
at a rate of 3 per cent (or more) of its body weight daily, would need
amino acids more in proportion to the needs for the formation of new
tissues; the amino acid requirements of the human child growing at a
rate of some 0.03 per cent of its body weight daily would be dominated
more by the needs for amino acids to replace endogenous losses. In the
former case, the daily requirement of protein, following the point of
inflection in the growth curve, would be expected to decrease in absolute
amount, as the daily growth increments decrease. In the latter case, the
amount of protein required per day may be expected to increase as the
body weight increases, but at a slower rate.
The amino acids needed for growth are needed mainly for the syn
thesis of the protein molecules entering into the structure of protoplasm.
For this function the simultaneous presence in the tissues of all of the
amino acids that the body cannot manufacture itself from dietary con
stituents is required. The absence of any one will block the synthetic
processes. The amino acids needed for maintenance are needed mainly
for the formation of creatine, carnosine, glutathione, ergothionine, thy-
roxine, adrenalin and other nitrogenous tissue constituents or tissue
products destroyed in metabolism. For these replacement functions
the assortment of amino acids needed is simple and will vary from one
type of synthetic reaction to the other. The absence from the diet of
any one essential amino acid will block one or more of these anabolic
reactions but not all. Hence, a dietary protein like gelatin or zein, en
tirely deficient in one or more of the amino acids that the body cannot
DIFFERENCES IN AMINO ΑΟΠ) REQUIREMENTS 3
manufacture itself, may be partially utilized in maintenance, as McCol-
lum and Steenbock (2) showed many years ago in metabolism experi­
ments on growing pigs. For growth, gelatin possesses a biological value
of zero, since no growth will occur with gelatin as the sole source of
amino acids. But for maintenance, it possesses a biological value of 25
(3), indicating that many of the replacement reactions of maintenance
do not need the amino acid entirely lacking from the gelatin molecule,
i.e., tryptophane. Maintenance will also involve some protein synthesis
for the growth of epidermal structures, for the replacement of red blood
cells destroyed during metabolism, and probably for many other pur­
poses, although the net aggregate in terms of nitrogen may not be large
in proportion to the total replacement aggregate.
An important recent development in amino acid nutrition is the dem­
onstration that the nonessential amino acids can be synthesized in the
body of the monogastric animal from glutamic acid and the unnatural
forms of the essential amino acids (4), from ammonium citrate (5), and
from glycine and urea (6). The ad libitum feeding technique employed
in the latter two reports detracts to some extent from the conclusiveness
which the interpretation of the nitrogen-balance data might otherwise
possess. The utilization of ammonia for amino acid synthesis (7, 8, 9),
but not that of urea (10, 11), has been confirmed by the use of appro­
priate dietary supplements labeled with the N15 isotope.
II. AN EXPERIMENTAL STUDY
The experiments that will be reported in this section were designed
to test the statements just made concerning the differences in amino acid
requirements that may be expected to accompany differences in age and
in species among animals. They were carried out upon young and upon
mature rats, not with different mixtures of amino acids, but with differ­
ent proteins known to differ in amino acid content and in nutritive
quality. The results obtained will be compared with those secured with
adult human subjects subsisting on the same protein sources.
Most of the data on human subjects will be taken from an article
published in 1948 from Murlin's laboratory at the University of Roch­
ester (12). Many of the data on growing and adult rats have been taken
from a paper by Mitchell and Beadles (13). Both series of experiments
were part of a cooperative study of protein utilization sponsored by the
Bureau of Biological Research of Rutgers University. The proteins, or
protein foods, used by all cooperating laboratories were the same, con­
sisting of egg albumin, desiccated and defatted whole egg dried and
y
defatted beef muscle, wheat gluten, casein and peanut flour.
4 H. H. MITCHELL
1. Experimental Methods
The experiments, both those at the University of Illinois and those at
the University of Rochester, were carried out in accordance with the
nitrogen balance method, yielding coefficients of true digestibility and
biological values. In the work on growing rats and in Murlin's work
on adult men and women, these measures of protein utilization in di
gestion and in metabolism were computed by the Thomas method, or a
modification of this method to adapt it to the growing animal. In the
work on adult rats, the calculations were made on the basis of data
secured with 3 dietary levels of each test protein; the biological values
were computed from the slope of the line describing the relationship of
absorbed nitrogen and of nitrogen balance, at levels of intake insufficient
to support nitrogen equilibrium or to induce any considerable nitrogen
retention (14), a modification of a method introduced by Melnick and
Cowgill (15). The data were expressed per calorie of basal metabolism
and were pooled together for each protein in computing regression
equations. The standard errors of the slopes of the regression lines were
computed by a method described by Rider (16).
2. Experimental Results
In Table I, the biological values of the nitrogen in the 6 protein
sources for growing rats, for mature rats and for humans are assembled
for comparison. The differences in digestibility, not given in the table,
are not great with one or two exceptions and their discussion does not
seem to be a profitable undertaking, since the factors determining the
completeness of protein digestion are so little understood.
TABLE I
The Biological Value of the Test Proteins by Growing Eats,
Mature Bats and Adult Men
Biological value
Growing Mature Mature
Protein rate rats humans
Egg albumin 97 94 91
Whole egg (commercial) 87 82 94
Beef muscle 76 69 67
Wheat gluten 40 65 42
Casein 69 51 56
Peanut flour 54 46 56
DIFFERENCES IN AMINO ACID REQUIREMENTS 5
3. Interpretations
The differences in biological value, however, may be correlated to
some extent with the amino acid composition of the proteins. Beef
muscle, casein and peanut flour are deficient in cystine or methionine or
both for the growing rat, while wheat gluten is markedly deficient in
lysine (3).
Prom the table it will be evident that for those proteins deficient in
cystine-methionine, i.e., beef, casein and peanut, the biological values are
lower for the adult rat than for the growing rat, and, with the exception
of beef, they are lower than for the adult human. The cystine-methio­
nine deficiency of beef muscle is comparatively small. On the other
hand, for wheat gluten, deficient in lysine, the biological value for the
adult rat is considerably higher than for the growing rat or for the adult
human.
With regard to egg albumin and the proteins of whole egg, no such
comparisons can be made among growing rats, mature rats and mature
humans, because no conclusive information is available concerning their
limiting amino acids. The estimations of Mitchell and Block (17), based
upon the comparative amino acid analyses of whole egg proteins and of
egg albumin would lead one to suspect a small lysine deficiency in the
latter protein.
4. A Test of the Amino Acid Adequacy of Whole Egg Proteins
The nutritive quality of whole egg proteins and of egg albumin is so
high that its limitation by amino acid deficiencies, if such exist, would be
expected to be slight. If whole egg proteins are a perfect protein mix­
ture for the growing rat, in the sense that they contain the essential
amino acids in the same proportions that exist among the requirements
for these amino acids, then supplementation of whole egg proteins with
individual essential amino acids should not improve their growth-pro­
moting value. In testing this proposition experimentally a whole egg
sample prepared in the laboratory by fat extraction and dehydration at
low temperatures was used. Samples prepared by this method have
yielded biological values for the growing rat of 94 to 96, and protein
eflScieney ratios (in a 28-day test) averaging 4.00 g. weight gain per g. of
protein consumed when fed at a dietary level of 9 to 10 per cent.
A basal diet was prepared containing approximately 9 per cent of
conventional protein (Νχ6.25) from defatted dehydrated whole egg
y
12 per cent of fat, 2 per cent of roughage and adequate additions of
minerals and vitamins. It was fed for 28 days to 12 groups of 8 wean-
6 H. H. MITCHELL
ling rats each to test the response in growth of supplements of individual
amino acids, a mixture of 9 essential amino acids, omitting arginine, and
of 3 per cent additional egg protein. Each group of rats was treated
as 4 pairs, one rat in each pair receiving the unsupplemented basal diet
and one receiving the respective supplement. The food intakes of pair
mates were equalized. The average results of the test are summarized
in Table II.
TABLE II
The Results of Amino Acid (or Protein) Supplementation of Laboratory-Prepared
Whole-Egg Proteins Fed at a 9-Per Cent Level in Paired-Feeding Tests
Average gain in body weight
after 28 days of feeding
Amino acid Concentration Number Without With
(or other) of supplement of supplement, supplement. , Difference,
supplement in diet pairs 9- 9- 9*
%
DL-Lysine 0.4 4 116.8 120.2 3.4
L-Histidine 0.2 4 91.7 92.7 1.0
L-Arginine 0.4 3* 74.3 76.0 1.7
DL-Phenylalanine 0.2 4 116.2 117.2 1.0
L(-Tryptophane 0.2 4 107.5 108.0 0.5
DL-Methionine 0.2 4 115.0 116.2 1.2
DL-Threonine 0.2 4 88.5 86.5 —2.0
DL-Leucine 0.4 4 114.0 112.8 — 1.2
DL-Isoleucine 0.6 3» 95.7 95.7 0
DL-Valine 0.2 4 117.8 121.5 3.7
9 essentials α 3.0 10 82.1 92.3 10.2
Additional egg prot eins* 3.0 8 79.0 88.6 9.6
a These tests continued for only 21 days. In all pairs the rat on the ι supplemented
diet gained at a faster rate and attained the greater body length.
6 The group was reduced to 3 pairs because of the occurrence of respiratory disease
in one rat of a pair.
The last two entries in the table demonstrate clearly that the 9 per
cent level of egg protein was inadequate to promote maximum growth:
in all pairs in each test the greater gain in body weight was made by the
rat receiving the supplement of 3 per cent either of the 9 essential amino
acids or of whole egg protein. The outcome of these tests prove that the
basal diet was suitable for the detection of individual amino acid defi­
ciencies.
The average growth responses induced by the individual amino acids
were small, though in 7 of the 10 cases they favored the supplemented
diet. If we consider the average differences between pair mates to be
entirely due to uncontrolled experimental conditions, they can be pooled.