The Effect of Fortification of processed Soya Flour with di-Methionine Hydroxy Analogue or di-Methionine on the Digestibility, Biological Value, And Net Protein Utilization of the Proteins as Studied in Children


Parthasarathy, H.N.; Doraiswamy, T.R.; Panemangalore, M.; Rao, M.N.; Chandrasekhar, B.S.; Swaminathan, M.; Sreenivasan, A.; Subrahmanyan, V.

Canadian Journal of Biochemistry 42: 377-384

1964


(a) Eight girls aged 8 to 9 years, height from 114.3 to 127.7 cm, weight from 17.0 to 23.8 kg, from a low income group in Mysore who were accustomed to a diet of cereals, millets and legumes received in 3 meals daily a basal low-protein diet, adequate in other nutrients. The test was of five 10-day periods in which the children received the basal diet with supplements, respectively, of 41 g full-fat soya flour, or soya flour and 1.2 g DL-methionine per 16 g N, or soya flour with the same amount of DL-methionine hydroxy analogue, or 56.8 g skimmed milk powder, or no supplement. Faeces and urine were collected during the second 5 days of each period; daily excretion of creatinine ranged from 457 to 511 mg and of creatine from 49 to 75. Estimations were made of total N in diet, urine and faeces, and of the essential amino acid intake on each diet, also of digestibility coefficients, biological value (BV) and net protein utilization (NPU). Average daily excretions of N in urine and faeces on the low-protein diet were 1.05 and 0.7 g, respectively. For the supplemented diets, in the order given, intakes were 3.91, 4.11, 4.11 and 4.13 g, excretion in urine 2.25, 1.95, 2.05, and 1.68 g, excretion in faeces 1.36, 1.28, 1.32 and 1.26 g. True digestibility of the proteins was 84, 86.4, 85.8 and 87.1, and BV was 63.5, 74.9, 71.5 and 82.6. Protein intakes in g per kg were 1.19, 1.25, 1.25 and 1.26 and NPU was 0.63, 0.81, 0.77 and 0.90, compared with 0.6 and 0.9 as the minimum and optimum by FAO reference protein requirements and 0.64 and 0.96 as the minimum and optimum ideal protein requirements in g per kg. Supplementation of soya flour with methionine caused a marked increese in BV and NPU; the hydroxy analogue was less effective. Values for both were significantly less than for skimmed milk powder.

THE
EFFECT
OF
FORTIFICATION
OF
PROCESSED
SOYA
FLOUR
WITH
dl-METHIONINE
HYDROXY
ANALOGUE
OR
dl-METHIONINE
ON
THE
DIGESTIBILITY,
BIOLOGICAL
VALUE,
AND
NET
PROTEIN
UTILIZATION
OF
THE
PROTEINS
AS
STUDIED
IN
CHILDREN
H.
N.
PARTHASARATHY,
T.
R.
DORAISWAMY,
MYNA
PANEMANGALORE,
M.
NARAYANA
RAO,
B.
S.
CHANDRASEKHAR,
M.
SWAMINATHAN,
A.
SREENIVASAN,
AND
V.
SUBRAHMANYAN
Central
Food
Technological
Research
Institute,
Mysore,
India
Received
September
10,
1963
Abstract
The
true
digestibility
coefficient,
biological
value,
and
net
available
protein
of
diets
based
on
processed
soya
flour
supplemented
with
dl-methionine
hydroxy
analogue
(MHA)
or
dl-methionine
(at
a
level
of
1.2
g/16
g
N)
have
been
deter-
mined
in
children
aged
8-9
years.
The
mean
daily
intake
of
protein
by
the
children
on
the
different
diets
was
maintained
at
a
level
of
about
1.2
g/kg
body
weight.
Supplementation
of
soya
flour
with
dl-methionine
brought
about
a
marked
increase
in
the
biological
value
and
net
protein
utilization
of
the
proteins.
MHA
was,
however,
slightly
less
effective
than
dl-methionine
in
this
respect.
The
bio-
logical
value
and
net
protein
utilization
of
the
different
proteins
were
as
follows:
soya
flour,
63.5
and
53.3;
soya
flour
+
MHA,
71.5
and
61.4;
soya
flour
methio-
nine,
74.9
and
64.7;
and
skim
milk
powder,
82.6
and
72.0.
Introduction
During
recent
years,
studies
have
been
carried
out
by
several
workers
on
the
use
of
oilseed
meals
and
legumes
as
supplements
to
human
diets
and
also
for
the
treatment
of
protein
malnutrition
in
children
(1-3).
Legume
proteins,
in
general,
are
deficient
in
methionine
(4).
It
has
been
shown
by
certain
workers
in
experiments
with
animals
that
fortification
of
legume
proteins
with
dl-
methionine
brings
about
a
marked
improvement
in
their
nutritive
value
(5,
6).
In
an
earlier
publication
from
this
laboratory,
it
was
reported
that
fortification
of
soya-bean
proteins
with
dl-methionine
hydroxy
analogue
(MHA)
increased
the
protein
efficiency
ratio
and
net
protein
utilization
in
albino
rats
almost
to
the
same
extent
as
that
obtained
with
dl-methionine
(7).
The
present
paper
describes
the
results
of
studies
with
children.
Experimental
Materials
Spray-dried
skim
milk
powder
of
good
quality
was
used.
Processed
full-fat
soya
flour
was
prepared
according
to
Narayana
Rao
et
al.
(8).
The
essential
amino
acid
composition
of
the
proteins
of
the
soya
flour,
skim
milk
powder,
and
the
low-protein
diet
was
determined
according
to
the
methods
used
by
Krishnamurthy
et
al.
(9).
The
mean
intakes
of
the
essential
amino
acids
from
the
different
diets
were
calculated
by
using
the
above
values.
The
sample
of
calcium
salt
of
dl-methionine
hydroxy
analogue
(90%
purity)
used
in
this
study
was
kindly
supplied
by
Monsanto
Chemical
Company,
Canadian
Journal
of
Biochemistry.
Volume
42
(1964)
377
378
CANADIAN
JOURNAL
OF
BIOCHEMISTRY.
VOL.
42,
1964
U.S.A.
The
product
was
reported
by
the
manufacturers
to
contain
78.8%
pure
acid.
dl-Methionine
(E.
Merck,
U.S.A.)
was
used
as
the
source
of
methionine.
The
required
quantities
of
soya-bean
flour
were
fortified
with
dl-methionine
or
MHA
at
a
level
of
1.2
g/16
g
N
by
dry
mixing
in
a
mechanical
mixer.
Subjects
The
subjects
were
eight
girls
aged
8-9
years
and
were
residents
of
a
boarding
home
in
Mysore
city.
The
ages,
heights,
and
weights
of
the
girls
are
given
in
Table
I.
All
of
them
belonged
to
the
low-income
groups
of
the
population
and
were
accustomed
to
consuming
diets
based
on
cereals,
millets,
and
legumes.
TABLE
I
Ages,
heights,
and
weights
of
the
children
at
the
beginning
of
the
test
Girl
No.
Age
(years)
Height
(cm)
Weight
(kg)
1
9
127.7
23.8
2
9
124.S
21.8
3
9
122.6
21.5
4
9
122.8
20.9
5
9
120.6
19.6
6
S
118.8
18.7
7
8
117.1
20.5
8
8
114.3
17.7
Diets
The
composition
of
the
low-protein
diet
used
in
this
study
is
given
in
Table
II.
This
diet
provided
1459
calories
and
contained
about
2.8
g
protein
(N
X
6.25).
The
children
received
the
low-protein
diet
throughout
the
experiment.
In
addition,
the
children
received
either
soya
flour
(with
or
without
added
d/-methionine
or
MHA)
or
skim
milk
powder
as
a
source
of
protein
during
the
first
four
periods
of
the
experiment.
They
were
fed
three
times
a
day,
i.e.
in
the
morning,
noon,
and
night.
The
mineral
salts
and
vitaminized
starch
were
mixed
with
the
tapioca
flour
and
corn
starch.
Tapioca
flour
was
given
in
the
form
of
unleavened
bread
while
the
corn
starch
was
made
into
sweet
and
savory
vermicelli-like
preparations.
In
addition,
the
children
received
a
vege-
table
soup
and
a
sweetened
drink
containing
ascorbic
acid.
Vitamins
A
and
D
were
added
to
the
vegetable
oil.
Full-fat
soya
flour
(41.0
g)
was
given
in
three
equal
doses
along
with
breakfast,
lunch,
and
dinner
in
the
form
of
sweet
pud-
ding.
Skim
milk
powder
(56.8
g)
was
also
given
in
three
equal
doses
(after
reconstitution
in
6
times
the
weight
of
water and
addition
of
cane
sugar)
along
with
the
three
meals.
The
mean
daily
intake
of
protein
oil
the
soya
flour
or
skim
milk
powder
diets
was
maintained
at
a
level
of
about
1.2
g/
kg
body
weight.
Feeding
of
Children
and
Collection
of
Urine
and
Faeces
The
metabolism
period
consisted
of
five
periods
of
10
days
each;
period
1,
PARTHASARATHY
ET
AL.:
PROTEIN
UTILIZATION
379
TABLE
II
Mean
daily
intake
(g)
of
foodstuffs
by
the
children
on
different
diets*t
Foodstuffs
Low-
protein
diet
Diets
based
on
Soya
flours
Skim
milk
powder
Basal
low-protein
diet
Tapioca
flour
(washed
with
dilute
alkali)
134.0 115.0
105.0
Corn
starch
120.0
91.0
91.0
Sugar
58.0
58.0
5S.0
Peanut
oil
(fortified
with
vitamins
A
and
D)
37.0
37.0
37.0
Salt
mixture§
5.0 5.0
5.0
Vitaminized
starchll
5.0
5.0
5.0
Supplements
Processed
full-fat
soya
flour
41.0
Skim
milk
powder
56.8
*All
the
diets
supplied
in
addition
(g/day):
common
salt,
8.0;
onion,
14.0;
tamarind
fruit
pulp,
5.0;
non-leafy
vegetables
(knolkhol,
brinjals,
ladies
finger,
and
radish
white),
60.0;
condiments
(red
chillies
and
mustard)
3.0.
The
protein
content
(N
X
6.25)
of
the
different
diets
were
as
follows:
low-protein
diet,
2.8
g;
soya
flour
diet,
24.4
g;
and
skim
milk
powder
diet,
25.8
g.
t
Each
child
received
50
mg
of
ascorbic
acid
daily
in
the
form
of
a
sweetened
drink.
Tin
the
2nd
and
3rd
period,
each
child
in
addition
was
given
260
mg
of
dl-methionine
or
methionine
hydroxy
analogue
respectively.
§Osborne
and
Mendel
salt
mixture.
11Provided
the
daily
requirements
of
B
vitamins
as
recommended
by
the
(U.S.A.)
National
Research
Council,
Food
and
Nutrition
Board
(1958).
soya
flour
diet;
period
2,
soya
flour
+
dl-methionine
diet;
period
3,
soya
flour
MHA
diet;
period
4,
skim
milk
powder
diet;
and
period
5,
low-protein
diet.
The
first
5
days
on
each
diet
were
treated
as
a
preliminary
period
for
the
children
to
get
accustomed
to
the
diet
and
the
collection
of
urine
and
faeces
was
confined
to
the
last
5
days
in
each
period.
Carmine
was
used
as
a
marker
for
the
collection
of
faeces.
The
daily
excretion
of
creatinine
in
urine
was
deter-
mined
as
a
check
for
the
quantitative
collection
of
urine.
The
daily
excretion
of
creatinine
in
the
subjects
ranged
from
457
mg
to
511
mg
per
day
and
of
creatine
48
to
75
mg
per
day.
In
the
same
subject
the
daily
excretion
of
creati-
nine
and
creatine
on
different
days
during
the
metabolism
period
did
not
differ
by
more
than
4%,
indicating
thereby
that
the
collection
of
urine
was
almost
quantitative.
Duplicate
samples
of
the
different
diets
consumed
daily
by
each
child
were
collected
and
dried
at
60-65°
C
in
a
cabinet
drier.
They
were
powdered
and
kept
in
glass-stoppered
bottles
for
analysis.
The
urine
and
faeces
were
preserved
according
to
Murthy
et
al.
(10).
Total
nitrogen
in
diet,
urine,
and
faeces
were
determined
by
the
micro-Kjeldhal
method.
The
pattern
of
diets
consumed
by
the
children
during
the
different
periods
is
given
in
Table
II.
The
essential
amino
acid
composition
of
the
diets
is
given
in
Table
III.
Data
regarding
the
amino
acid
intake
of
children
on
the
different
diets,
as
compared
with
children's
amino
acid
requirements
as
reported
by
Nakagawa
et
al.
(11-
14),
are
given
in
Table
IV.
The
digestibility
coefficient,
biological
value,
net
protein
utilization
(NPU)
380
CANADIAN
JOURNAL
OF
BIOCHEMISTRY.
VOL.
42,
1964
TABLE
III
Essential
amino
acid
content
(g/16
g
N)
of
the
mixed
proteins
of
different
diets
Amino
acid
Diets
based
on
FAO
Ideal
reference
protein
pattern
(16)
Soya
flour
Soya
flour
+
methionine
or
MHA
Skim
milk
powder
reference
protein
pattern
(15)
Arginine
7.3
7.3
4.1
6.6
Histidine
2.6
2.6
2.3
-
2.4
Lysine
6.6
6.6
7.8
4.2
7.5
Leucine
7.7
7.7
9.9
4.8
10.0
Isoleucine
5.5
5.5
6.7
4.2
6.6
Methionine
1.5
1.5
+
1.2*
2.4
2.2
2.8
Cystine
1.7 1.7
0.9
-
2.0
Total
sulphur
amino
acids
3.2
f$
3.2
+
1.2*
3.3
f$
4.2
4.8
Phenylalanine
5.0
5.0
5.5
2.8
5.8
Threonine
3.9
3.9t
4.5
2.8
5.0
Tryptophan
1.3
1.3f
1.4 1.4
1.6
Valine
5.3
5.3
5.0
4.2
7.0
Protein
score
f
67
78
68
-
100
76
93
79
100
*Methionine
or
MHA.
tAmino
acids
limiting
as
compared
to
FAO
pattern.
Amino
acids
limiting
as
compared
to
Ideal
reference
protein
pattern.
TABLE
IV
Mean
daily
intake
(mg/kg)
of
essential
amino
acids
by
the
children
from
the
different
diets
as
compared
with
the
amino
acid
requirements
Amino
acid
Basal
low-protein
diet
Diets
based
on
Amino
acid*
require-
ments
Soya
flour
Soya
flour
+
methionine
or
MHA
Skim
milk
powder
Arginine
6.8
86.5
91.3
51.5
Histidine
1.9
31.3
33.0
28.7
Lysine
4.4
77.8
82.2
97.1
60
Leucine
10.2
92.0
96.9
124.1
45
Isoleucine
5.3
65.1
68.7
84.6
30
Methionine
1.9
17.6
18.6
+
13.3t
29.8
27
Cystine
1.5
20.4
21.5
11.6
Total
sulphur
amino
acids
3.4
38.0
40.1
+
13.31*
41.4
Phenylalanine
5.8
59.3
62.5
68.3
27
Threonine
4.4
46.3
48.9
55.8
35
Tryptophan
1.5
15.1
16.0
17.1
9
Valine
8.2
62.7
66.0
62.9
33
*Data
of
Nakagawa
et
al.
(11-14).
tMethionine
or
MHA.
and
net
available
protein
were
calculated
according
to
the
following
formulae:
Apparent
digestibility
coefficient
=
100
X
N
intake
-
faecal
N
N
intake
N
intake
N
intake
-
(faecal
N
-
endogenous
True
digestibility
coefficient
=
100
X
faecal
N)
PARTHASARATHY
ET
AL.:
PROTEIN
UTILIZATION
381
N
intake
(faecal
N
endogenous
faecal
N)
true
digestibility
coefficient
X
biological
value
100
Net
available
protein
protein
intake
X
NPU
(o
p
)
100
Statistical
Treatment
of
Data
The
data
were
analyzed
by
the
analysis
of
variance
method
appropriate
for
randomized
block
design,
considering
each
subject
as
a
block
and
differences
tested
for
significance
by
using
a
one-sided
or
two-sided
t
test,
whichever
is
appropriate.
Results
Data
regarding
the
daily
urinary
and
faecal
endogenous
nitrogen
on
the
low-
protein
diet
are
given
in
Table
V.
The
mean
daily
balance
of
nitrogen,
digesti-
bility
coefficient,
biological
value,
and
net
protein
utilization
of
the
protein
in
children
fed
on
diets
based
on
soya
(with
or
without
added
MHA
or
dl-methio-
nine)
or
on
skim
milk
powder
is
given
in
Table
VI.
The
net
available
protein
on
the
different
diets
is
given
in
Table
VII.
TABLE
V
Daily
urinary
and
faecal
excretion
(g)
of
nitrogen
by
the
children
on
the
low-protein
diet
Girl
No.
Urinary
Faecal
Total
1
1.14
0.78
1.92
2
1.14
0.72
1.86
3
1.05
0.75
1.80
4
1.05
0.74
1.79
5
0.98
0.76
1.74
6
0.98
0.76
1.74
7
0.99
0.71
1.70
8
1.06
0.63
1.69
Mean
value
with
its
standard
error
(7
d.f.)
1.05±0.023
0.73±0.016
1.78±0.028
Essential
Amino
Acid
Intake
and
Requirements
(Tables
III
and
IV)
The
protein
scores
of
the
different
diets
as
compared
with
FAO
reference
protein
pattern
and
Ideal
reference
protein
pattern
(16)
calculated
according
to
the
method
of
FAO
Committee
(15)
are
as
follows:
soya
flour
diet,
76
and
67;
soya
flour
+
methionine
or
MHA
diet,
93
and
78;
and
milk
diet,
79
and
68
respectively.
Data
regarding
the
essential
amino
acid
intakes
and
requirements
of
the
children
are
given
in
Table
IV.
It
is
evident
that
soya-bean
protein
at
a
level
of
1.2
gjkg
body
weight
provided
the
essential
amino
acid
requirements
of
children
as
assessed
by
Nakagawa
et
al.
(11-14)
even
after
allowance
is
made
for
the
loss
of
16%
of
the
protein
in
digestion.
N
intake
(faecal
N
endogenous
faecal
N)
Biological
value
=
100
X
(urinary
N
endogenous
urinary
N)
NPU
(o
p
)
TABLE
VI
Mean
daily
balance
of
nitrogen
and
digestibility
coefficient,
biological
value,
and
net
protein
utilization
of
the
proteins
of
diets
based
on
soya
flour
supplemented
with
MHA
or
dl-methionine
Intake
Excretion
(g)
Balance
Appa-
rent
digesti-
bility
True
digesti-
bility
Bio-
logical
value
NPU
(c
,p
)
%
mg/kgintake
Diets*
gmg/kg
Urinary
Faecal
Total
Soya
flour
3.91
190
2.25
1.36
3.61
0.30
15.1
7.7
65.3
84.0
63.5
53.3
Soya
flour
+
methionine
4.11
200
1.95
1.28
3.23
0.88
43.5
21.4
68.7
86.4
74.9
64.7
Soya
flour
+
MHA
4.11
200
2.05
1.32
3.37
0.74
36.3
18.1
68.0
85.8
71.5
61.4
Skim
milk
powder
4.13
200
1.68
1.26
2.94
1.19
58.6
28.8
69.4
87.1
82.6
72.0
Standard
error
of
the
mean
(21
d.f.)
±0.03
±1.67
±0.75
±0.86
±0.76
±0.90
±0.75
*Calorie
value:
1460
kcal.
TABLE
VII
Mean
protein
intake
and
net
available
protein
in
children
on
different
diets
FAO
reference
protein
requirementst
Protein
intake
Net
available
protein*
(g/kg)
Ideal
protein
requirementst
(g/kg)
Diet
g
g/kg
g
g/kg
Minimum
Optimum
Minimum
Optimum
Soya
flour
Soya
flour
+
methionine
Soya
flour
+
MHA
Skim
milk
powder
24.4
25.7
25.7
25.8
1.19
1.25
1.25
1.26
13.0
16.6
15.8
18.6
0.63
0.81
0.77
0.90
0.6
0.90
0.64
0.96
*(Protein
intake
X
NPU)
÷
100.
tFAO
rept.
No.
16.
FAO,
Rome,
1957.
M.
Swaminathan.
Indian
J.
Pediat.
30,
189
(1963).
CAD
00
t•0
C
A
NADIA
N
JO
UR
NAL
OF
B
I
OC
H
EM
IS
T
RY
.
PARTHASARATHY
ET
AL.:
PROTEIN
UTILIZATION
383
Nitrogen
Balance
in
Children
and
Digestibility
Coefficient
and
Biological
Value
of
the
Proteins
(Table
VI)
The
mean
daily
N
intake
from
the
different
diets
ranged
from
:3.91
to
4.13
g
(about
200
mg/kg
body
weight).
The
mean
daily
N
retention
ranged
from
0.30
g
on
soya
flour
diet
to
1.19
g
on
milk
diet
(15.1
mg
to
58.6
mg/kg
body
weight).
The
mean
true
digestibility
coefficient
of
the
proteins
ranged
from
84.0
to
86.4
on
the
soya
flour
diet
and
the
same
fortified
with
dl-methionine
and
MHA
as
compared
with
87.1
for
milk
diet.
The
biological
value
of
soya
proteins
was
63.5,
which
significantly
increased
(P
<
0.001)
to
74.9,
when
fortified
with
dl-Methionine
and
to
71.5
(P
<
0.001)
when
fortified
with
MHA.
dl-Methio-
nine
hydroxy
analogue
was,
however,
significantly
less
effective
(P
<
0.01)
than
dl-methionine
in
increasing
the
biological
value
of
soya
proteins.
Net
Protein
Utilization
and
Net
Available
Protein
(Tables
VI
and
VII)
The
NPU
(0
)
of
diet
based
on
soya
flour
+
MHA
(61.4)
was
significantly
less
(P
<
0.01)
than
that
of
a
diet
based
on
soya
flour
+
dl-methionine
(64.7),
which
in
turn
was
significantly
less
(P
<
0.001)
than
that
(72.0)
of
milk
proteins.
The
net
available
protein
(g/kg
body
weight)
from
the
different
diets
were
as
follows:
soya
flour,
0.63;
soya
flour
+
dl-methionine,
0.81;
soya
flour
+
MHA,
0.77;
and
skim
milk
powder,
0.90
as
compared
with
FAO
reference
protein
requirements
(15)
of
0.6
g
(minimal)
and
0.9
g
(safe
practical
allowance)
and
Ideal
reference
protein
requirements
of
0.64
g
(minimal)
and
0.96
(optimal)
suggested
by
one
of
us
(16).
Discussion
The
results
obtained
in
the
present
study
with
children
have
shown
that
fortification
of
soya
flour
with
dl-methionine
or
dl-methionine
hydroxy
analogue
(MHA)
(at
a
level
of
1.2
g/16
g
N)
brings
about
a
significant
increase
in
the
biological
value
and
net
protein
utilization
of
the
proteins.
dl-Methionine
hydroxy
analogue,
however,
was
significantly
less
effective
than
dl-methionine
in
this
respect.
The
biological
value
and
net
protein
utilization
of
soya
protein
fortified
with
d/-methionine
or
MHA
were
significantly
less
than
those
of
milk
proteins.
Studies
reported
earlier
with albino
rats,
however,
showed
that
supplementation
of
soya
flour
with
dl-methionine
or
MHA
increased
the
pro-
tein
efficiency
ratio
and
net
protein
utilization,
almost
to
the
same
extent
as
those
of
milk
proteins
(7).
The
results
obtained
with
children
in
the
present
study,
therefore,
differ
to
some
extent
from
those
obtained
with
albino
rats.
The
mean
daily
intake
of
protein
was
maintained
at
a
level
of
about
1.2
g/kg.
The
net
available
protein
from
soya
flour
diet
(0.63
g)
was
nearly
equal
to
the
`minimal'
protein
requirements
as
FAO
reference
protein
(0.6
g)
or
Ideal
reference
protein
(0.64
g)
but
less
than
the
'optimal'
requirements
(0.9
and
0.96
g
respectively).
Supplementation
of
soya
flour
with
MHA
or
dl-methionine
increased
the
net
available
protein
to
0.77
g
and
0.81
g/kg
respectively,
as
compared
with
a
value
of
0.90
obtained
for
milk
proteins.
Even
though
the
results
obtained
with
children
in
the
present
short
term
study
have
shown
that
384
CANADIAN
JOURNAL
OF
BIOCHEMISTRY.
VOL.
42,
1964
MHA
is
an
effective
supplement
to
soya
proteins
deficient
in
methionine,
there
is,
nevertheless,
need
for
conducting
long-term
studies
with
albino
rats
and
other
animals
and
also
with
human
subjects
to
ascertain
whether
NI
HA
will
be
as
effective
as
dl-methionine
as
a
supplement
to
proteins
deficient
in
sulphur
amino
acids
over
long
periods
of
feeding.
Acknowledgment
We
are
thankful
to
Dr.
Arnold
E.
Shaefer
for
kindly
arranging
supplies
of
MHA
used
in
the
experiment.
This
work
was
supported
by
P.L.480
funds
from
the
U.S.
Public
Health
Service.
References
1.
V.
SUBRAHMANYAN,
M.
NARAYANA
RAO,
and
M.
SWAMINATHAN.
Proc.
Natl.
Inst.
Sci.
India
Pt.
A,
26
(Suppl.
1),
99
(1960).
2.
N.
S.
SCRIMSHAW
and
R.
BRESSANI.
Federation
Proc.
20
(Suppl.
7),
80
(1961).
3.
M.
AUTRET
and
A.
G.
VAN
VEEN.
Am.
J.
Clin.
Nutr.
3,
234
(1955).
4.
S.
KUPPUSWAMY,
M.
SRINIVASAN,
and
V.
SUBRAHMANYAN.
Proteins
in
foods.
Indian
Council
Med.
Res.
Spec.
Rept.
No.
33.
I.C.M.R.,
New
Delhi.
1958.
5.
K.
JOSEPH,
M.
NARAYANA
RAO,
M.
SWAMINATHAN,
K.
INDIRAMMA,
and
V.
SUBRAHMAN-
VAN.
Ann.
Biochem.
Exptl.
Med.
Calcutta,
20,
243
(1960).
6.
W.
C.
RUSSELL,
M.
W.
TAYLOR,
T.
G.
MEHRHOFF,
and
R. R.
HIRSCH.
J.
Nutr.
32,
313
(1946).
7.
H.
N.
PARTHASARATHY,
K.
JOSEPH,
M.
NARAYANA
RAO,
M.
SWAMINATHAN,
A.
N.
SAN-
KARAN,
A.
SREENIVASAN,
and
V.
SUBRAHMANYAN.
J.
Nutr.
Dietet.
1,
14
(1964).
8.
M.
NARAYANA
RAO,
T.
K.
ANANTHACHAR,
K.
R.
KURUP,
R.
RAJAGOPALAN,
M.
SWAMINA-
THAN,
A.
SREENIVASAN,
and
V.
SUBRAHMANYAN.
J.
Nutr.
Dietet.
1,
1
(1964).
9.
K.
KRISHNAMURTHY,
P.
K.
TASKER,
T.
N.
RAMAKRISHNAN,
R.
RAJAGOPALAN,
and
M.
SWAMINATHAN.
Ann.
Biochem.
Exptl.
Med.
Calcutta,
20,
73
(1960).
10.
H.
B.
N.
MURTHY,
M.
SWAMINATHAN,
and
V.
SUBRAHMANYAN.
Brit.
J.
Nutr.
8,
11
(1954).
11.
I.
NAKAGAWA,
T.
TAKAHASHI,
and
T.
SUZIKI.
J.
Nutr.
71,
176
(1960).
12.
I.
NAKAGAWA,
T.
TAKAHASHI,
and
T.
SUZIKI.
J.
Nutr.
73,
186
(1961).
13.
I.
NAKAGAWA,
T.
TAKAHASHI,
and
T.
SUZIKI.
J.
Nutr.
74,
401
(1961).
14.
I.
NAKAGAWA,
T.
TAKAHASHI,
T.
SUZIKI,
and
K.
KOBAYASHI.
J.
Nutr.
77,
61
(1962).
15.
FAO
Nutr.
Studies
No.
16.
Protein
Requirements.
Food
Agr.
Organ.
U.N.,
Rome.
1957.
16.
M.
SWAMINATHAN.
Indian
J.
Pediat.
30,
189
(1963).