Journal of Cereal Science 33 (2001) 183–191

doi:10.1006/jcrs.2000.0355, available online at http://www.idealibrary.com on

Comparative Protein Digestibility in Growing Rats of

Cooked Rice and Protein Properties of Indica and
Japonica Milled Rices
Sigurd Boisen∗, Joy Bartolome A. Duldulao†, Evelyn Mae T. Mendoza‡ and
Bienvenido O. Juliano§

∗ Danish Institute of Animal Science, Department of Nutrition, Research Centre Foulum, DK-8830 Tjele,
Denmark; † Philippine Rice Research Institute, Maligaya, 3119 Muñoz, Nueva Ecija, Philippines;
‡ University of the Philippines Los Baños, Biochemistry Laboratory, Institute of Plant Breeding,
4031 College, Laguna, Philippines; § Philippine Rice Research, Institute Los Baños, 4031 College,
Laguna, Philippines.

Received 1 June 1999

ABSTRACT
Eight indica and japonica milled rices with low amylose content and low starch gelatinisation
temperature were analysed for cooked rice energy and N balance in growing rats and for protein
properties. Digestible energy values were similar. Japonica rices Koshihikari and Sasanishiki had
higher true digestibility (TD) and net protein utilisation in rats than indica rices IR24 and PR23383-
15. Similar results were obtained from in vitro proteolysis. High TD of cooked rices was not significantly
correlated with low prolamin content in raw rice and with low waxy gene product (protein bound
to starch granule) but was significantly correlated with low cysteine content in protein and with low
denatured prolamin content in cooked rice. In two pairs of cooked waxy milled rices, which are
devoid of waxy gene product, japonica rice still tended to show higher rat TD and in vitro protein
digestibility than indica rice, consistent with lower cysteine content in japonica protein.
 2001 Academic Press

Keywords: cooked-rice protein digestibility, indica vs. japonica, protein properties, waxy gene product,
cysteine content, protein content, glutelin content, prolamin content.

INTRODUCTION to be the core proteins of the prolamin-rich PB

The storage protein bodies (PB) in the rice en-
dosperm are the prolamin-rich spherical PB I and
the glutelin-rich segmented crystalline PB II1. True
protein digestibility (TD) of milled rice in growing
rats have been shown2 to be reduced from 100%
to 85–90% by cooking the raw rice in boiling
water. The poorly digestible fraction was shown
Corresponding author: B. Juliano. Tel: (63-49) 536-3633; Fax:
(63-49) 536-3515; E-mail: bjuliano@laguna.net. S. Boisen and
J. B. A. Duldulao are the senior authors.
I3,4. But indica and japonica rices are reported to
have similar (85–90%) TDs3,5 in man.
Collaborative N balance studies in growing rats
with the late Dr Bjørn O. Eggum of the Danish
Institute of Animal Science (DIAS) involved 12
indica cooked rices (TD 83·3–97·4%) up to 19922,4,6.
Higher TD (94·6–99·8%) were obtained with more
recent studies on seven japonica cooked rices9,10,
but there was a tendency for TD to decrease
with increase in amylose content (AC, r=−0·76,
p<0·05). The TD of the combined 19 cooked
milled rices correlated both with protein content
(r=−0·83, p<0·01) and AC (r=−0·68, p<0·01).

0733–5210/01/030183+09 $35.00/0  2001 Academic Press

184 S. Boisen et al.
The waxy gene product (granule-bound starch
synthase) of japonica and indica nonwaxy rice
differ only by 16 individual nucleotides11. Indica
rice starch has more than twice as much 60-kD
subunit waxy gene product12,13 as japonica rice
starch, reflecting lower specific activity of the gran-
ule-bound starch synthase14. The starch synthase
is relatively stable and extractable only after starch
gelatinisation. Interestingly, high-AC indica rice
in Taiwan has 2% higher protein content than
low-AC japonica rice15.
Dr K. Tanaka of Kyoto Prefectural University
has reported (unpubl., 1985) on the higher in vitro
pepsin digestibility of protein in raw rice in three
japonica (78–80%) than in three indica rices (68,
69 and 78%). Protein contents were 7·4–8·8% for
japonica and 7·5–10·4% for indica rices. The
protein of japonica rice has been reported to be
20% prolamin, but indica rice protein has 30%
prolamin16. The prolamin and glutelin fractions
of milled rice protein increase with an increase in
grain protein content17.
Because of these considerations, a collaborative
study was undertaken on cooked milled rice at
similar low AC to determine; (1) whether or not
japonica rice protein has greater TD in growing
rats than indica rice protein; (2) whether the
difference is related to the increase in prolamin
content with an increase in protein content of
milled rice; and (3) whether this difference is in
any way related to the greater amount of waxy
gene product (granule-bound starch; synthase) in
indica rice.
MATERIALS AND METHODS
Materials
Brown rice samples of low AC and low starch
gelatinisation temperature (GT) were obtained
from the 1996 crop from various sources: two
popular Japanese japonica rices, Koshihikari and
Sasanishiki from Masao Yokoo, National Agri-
cultural Research Center, Tsukuba, Japan; four
Philippine rices: IR24, Sinandomeng, PR23383-
15, and PRJ 3 from University of the Philippines
Los Baños (UPLB)-PhilRice Los Baños and Phil-
Rice Maligaya; Thai variety Khao Dawk Mali 105
from Ngamchuen Kongseree, Pathumthani Rice
Research Center, Thanyaburi, Pathumthani,
Thailand; and U.S. long-grain variety Toro 2 from
Anna M. McClung, ARS-USDA Rice Research,
Beaumont, TX, USA. All had low AC (15–20%)
and low GT (alkali spreading value 6–7).
Malagkit Sungsong and IR29 low GT waxy
rices were obtained early in 1999 from PhilRice
Maligaya. Taikeng Glu. 5 and Taichung Sen
Glu. 1 low-GT waxy milled rices were obtained
from Sheu-chih Sheng, Taichung District Agri-
cultural Improvement Station, Tatsuen Hsiang,
Changhua, Taiwan.
Methods
Chemical studies on protein of these cooked and
raw milled rices were undertaken at the Institute
of Plant Breeding by J. B. A. Duldulao as his M.
S. thesis at UPLB. Indica-japonica classification
was checked by the seedling isozyme method18 at
the Isozyme Laboratory of D. S. Brar at the
International Rice Research Institute, and on waxy
gene product content of the rice starch13. Rice
flour (40 mg) was purified by five protein ex-
tractions with buffer A (55 m Tris HCl pH 6·8
with 2% (w/v) SDS, 5% (w/v)
-mercaptoethanol
and 10% (v/v) glycerol). The resulting starch gran-
ules were boiled in 10 volumes of buffer A for
10 min. After cooling, 10 volumes of buffer were
added and the slurry was centrifuged. A 20-L
supernate was used for SDS-PAGE. Crude protein
content was determined by micro-Kjeldahl
method using the factor 6·25 or by Lowry method.
Prolamin, albumin-globulin and glutelin contents
were determined on raw and cooked milled rice
flour by sequential extraction of 400 mg flour for
2 h each with 8 mL of 60% 1-PrOH, 0·7  NaCl
(pH 7·5), and 3% (w/w) SDS and 0·5% di-
thiothreitol (pH 8)19.
Rough and brown rices that pass the chemical
studies at PhilRice were milled after at least 4
months after harvest (aged), washed and boiled in
1·3 times its weight in water, freeze-dried and
airshipped to DIAS. At DIAS, the eight samples
were run together for amino acid analysis10, di-
gestible energy (DE), true digestibility (TD), bio-
logical value (BV), and net protein utilisation
(NPU) in five growing rats each10 and their in vitro
protein (EDN) and dry matter enzymic digestibility
(EDDM) values at the ileal level20 and in vitro
organic matter enzymic digestibility (EDOM) at
the faecal level21. Standard deviation (S.D.) was
0·2 g/16 g N for lysine, 0·1 g/16 g N for cysteine
and methionine10, 2% for in vitro EDN20, and 5%
for in vitro EDDM20 and EDOM21. Amino acid
 Rice protein digestibility and properties 185

Table I Classification of eight milled rices in terms of indica-japonica and starch properties (PhilRice Los Baños)

Propertya Koshihikari Sasanishiki Toro Sinandomeng PRJ Khao PR23383-15 IR24

2 3 Dawk
Mali
105

Glaszmann’s group VI VI 0 I I I I I
Waxy gene product Wxb Wxb Wxb Wxb Wxb Wxb Wxa Wxa
Waxy gene protein 0·07 0·06 0·11 0·08 0·06 0·08 0·13 0·20
(% dry basis)
Apparent amylose 15·2 15·5 14·0 13·0 11·8 11·1 14·8 13·0
(% dry basis)
Alkali spreading value 7·0 7·0 6·0 5·7 7·0 6·3 4·1 7·0
a
Glaszmann’s18 group: 1 (indica), group VI ( japonica); waxy gene product: Wxa (indica), Wxb ( japonica).

score was calculated based on the FAO/WHO/
UNU amino acid pattern for the preschool child
with lysine content of 5·8% as 100%22. Amino
acid score X TD was also calculated23.
Part of the raw milled rice was tested for alkali
spreading value24 as index of starch GT and for
AC by iodine colorimetry25.
Protein extracts and fractions and waxy gene
product were dispersed in buffer A in a boiling
water bath for 2 min and subjected to SDS-10%
gel PAGE26 with protein standards lysozyme
(14·3 kD), trypsinogen (24 kD), pepsin (34·7kD),
ovalbumin (45 kD), and bovine serum albumin
(66 kD). Polyacrylamide gels used were 0·75 mm
thick, 8 cm long and 7 mm wide. Gels were stained
with Coomassie blue and protein bands quantified
by QuantiScan software (Biosoft).
In vitro protein TD was also determined at UPLB
on raw and cooked milled rice flours20. Residual
proteins after pepsin and pancreatin digestion were
extracted with cold buffer A (residual proteins I)
and with boiling buffer A (residual proteins II) and
the extracts also subjected to SDS-PAGE. S.D.
were calculated and some data were subjected to
Duncan’s multiple range test27.
RESULTS AND DISCUSSION
Classification into indica and japonic types
The eight rices were all verified to have low AC
and low GT (Table I). An exception was the
mixture of high-GT grains (5 of 12) in PR23383-
15. One out of 12 grains had high GT in Si-
nandomeng. The rices had their expected Glasz-
mann18 grouping of I for indica and VI for
japonica, except for Toro 2 which could not be
classified (group 0). Khao Dawk Mali 105 was in
group I, although an earlier sample gave group
VI28. Classification by waxy gene product in the
endosperm using purified rice starch showed high
waxy gene product (indica, Wxa) only for
PR23383-15 and IR24 and Wxb ( japonica) for
the other six samples, including Khao Dawk Mali
105 and Toro 2, in conformity with previous
results28. The presence or absence of the 3 subunit
of glutelin28 could not be resolved from the SDS-
PAGE and could not be used for indica-japonica
classification28.
Aminogram and balance in rats
The freeze-dried cooked milled rices had 91·3–
93·2% dry matter or 6·8–8·7% moisture (Table
II). Protein content range was 6·19–10·87% and
was lower in japonica than in indica rices. Ash
content was 2·87–5·09%. Energy content was
17·5–18·4 J/g dry matter.
The amino acid values were used without cor-
rection for N recovery since the N recovery was
already high at 88·9–93·4% (Table II). Lysine was
the first limiting amino acid (3·1–3·4 g/16 g N) of
rice protein corresponding to an amino acid score
of 53–59%. Koshihikari and Sasanishiki protein
had the highest lysine content and amino acid
score. IR24, PR23383-15 and PRJ 3 proteins had
the lowest lysine value. Other essential amino acids
were present in amounts closer to the reference
pattern. However, Koshihikari and Sasanishiki
had lower cysteine content of protein than the
other rices and had lower tyrosine content than
IR24 and PR23383-15. Koshihikari protein has
the highest phenylalanine, arginine and leucine
levels.
Digestible energy (DE) in growing rats was sim-
186 S. Boisen et al.

Table II Properties, aminogram, rat balance data and in vitro digestibility of japonica and indica cooked milled rices
(Danish Institute of Animal Science)

Property Koshihikari Sasanishiki Toro Sinandomeng PRJ Khao PR23383-15 IR24

2 3 Dawk
Mali
105

Dry matter (%) 93·2 92·7 91·8 91·3 92·6 92·3 92·0 91·8
Protein (% N×6·25) 6·19 6·47 8·38 7·82 9·41 10·87 8·63 8·97
Crude ash (%) 2·98 2·87 2·98 3·20 5·09 3·09 4·51 3·44
Gross energy ( J/g 17·5 17·8 18·2 18·3 18·4 18·3 18·3 18·2
dry
matter)
Lys (g/16 g N) 3·4 3·4 3·2 3·2 3·1 3·2 3·1 3·1
Met (g/16 g N) 2·4 2·4 2·7 3·1 2·7 2·3 2·7 2·7
Cys (g/16 g N) 2·0 2·0 2·1 2·3 2·2 2·1 2·3 2·2
Thr (g/16 g N) 3·3 3·2 3·3 3·4 3·3 3·2 3·3 3·3
Ile (g/16 g N) 4·4 4·2 4·1 4·1 4·3 4·3 4·1 4·2
Leu (g/16 g N) 8·1 7·7 7·4 7·4 7·9 7·8 7·6 7·7
Val (g/16 g N) 6·1 5·9 5·8 5·8 5·9 6·0 5·9 5·9
Phe (g/16 g N) 5·3 4·9 4·9 5·0 5·1 5·1 5·1 5·1
Tyr (g/16 g N) 3·7 3·5 3·8 3·7 4·2 4·3 4·1 4·0
Arg (g/16 g N) 8·3 7·9 7·9 7·4 7·9 8·0 7·6 7·5
His (g/16 g N) 2·5 2·4 2·5 2·3 2·4 2·4 2·4 2·3
Ala (g/16 g N) 5·5 5·2 5·1 5·2 5·3 5·2 5·2 5·3
Asp (g/16 g N) 8·9 8·5 8·5 8·1 8·2 8·4 8·1 8·1
Glu (g/16 g N) 17·3 16·5 16·4 16·7 17·6 17·3 17·0 17·2
Gly (6/16 g N) 4·5 4·3 4·2 4·1 4·1 4·1 4·1 4·1
Pro (g/16 g N) 4·7 4·4 4·4 4·4 4·7 4·5 4·6 4·4
Ser (g/16 g N) 5·3 5·0 4·9 4·7 5·0 5·0 4·9 4·9
N recovery (%) 93·9 89·8 89·0 88·4 91·5 91·7 89·9 89·6
Amino acid scorea 59 59 55 55 53 55 53 53
(%)
Rat balance data
Digestible energy 97·4 97·5 97·4 97·2 96·9 97·2 96·9 97·0
(% of intake)±SD ±0·5 ±0·4 ±0·4 ±0·6 ±0·7 ±0·7 ±0·2 ±0·3
True digestibility 98·8 98·6 97·2 94·6 93·2 94·1 92·5 92·6
(% of N intake)±SD ±2·5 ±2·0 ±1·7 ±3·1 ±3·3 ±3·1 ±1·1 ±1·8
Biological value (% 77·3 81·5 70·6 75·8 73·9 72·5 74·1 71·7
of absorbed N)±SD ±3·1 ±2·0 ±2·5 ±3·1 ±3·1 ±2·1 ±3·4 ±2·5
NPU (% of N intake) 76·3 80·3 68·2 71·6 68·9 68·2 68·5 66·5
±SD ±3·9 ±1·7 ±1·4 ±1·6 ±4·8 ±1·7 ±2·9 ±3·4
Amino acid score X
TD (%) 58 58 54 52 50 52 52 49
EDNb (% at ileal 90·0 88·5 87·1 82·3 82·6 85·0 83·3 81·5
level)
EDDMb (% at ileal 82·8 77·4 84·5 80·1 78·3 79·4 71·7 76·7
level)
EDOMb (% at faecal 98·3 98·3 98·1 98·0 97·6 97·5 97·2 97·6
level)
a
Based on 5·8% lysine in FAO/WHO/UNU22 pattern as 100%.
b
EDN means enzyme digestible N (protein) at ileal level; EDDM means enzyme digestible dry matter at ileal level;
EDOM is enzyme digestible organic matter at faecal level.

ilar at 96·9–97·5% of intake (Table II). TD was
92·5–98·8% and highest for Koshihikari, Sa-
sanishiki and Toro 2, as in EDN values. PR23383-
15 and IR24 had lower TD than japonica rices.
Koshihikari and Sasanishiki had the lowest cyst-
eine content in protein. Amino acid digestibility
in cooked milled has been shown to be lowest for
cysteine2, suggesting disulphide bond formation.
Biological value was 70·6–81·5% and was highest
for Sasanishiki and lowest for Toro 2 and IR24.
 Rice protein digestibility and properties
NPU was 66·5–80·3% and highest for Sasanishiki
and lowest for IR24. Rat TD correlated positively
with NPU and lysine level in protein and negatively
with cysteine and tyrosine levels in protein (Table
IV).
Amino acid score corrected for TD was highest
for Koshihikari and Sasanishiki and lowest for
IR24 and PRJ 3 (Table II). Toro 2 had inter-
mediate value.
In vitro digestibility
In vitro EDN at the ileal level was 81·5–90·0% and
was highest for the two japonica samples and Toro
2 and lowest for IR24. In vitro EDDM at the ileal
level was 71·7–84·5%, highest for Toro 2 and
lowest for PR23383-15, and did not follow TD in
rats (Table II). EDOM at the faecal level was high
at 97·2–98·3% and was similar to DE values in
rats. In vitro digestible protein (DP) of raw milled
rice was 81·1–89·2% (Table III). It was also highest
for Koshihikari and Sasanishiki, followed by Toro
2. By contrast, TD of raw rice protein in growing
rats was 100%2. Corresponding DP values for
cooked milled rice were lower at 77·0–82·6%.
IR24 had the lowest value in both sets, whereas
japonica rices, Koshihikari and Sasanishiki, and
Toro 2 had the highest values. Cooked rice DP
values were less than the EDN values reported in
Table II, probably due to differences in the batches
of proteases used. Undigested proteins in raw rice
were soluble in cold (residual proteins I) and boiling
buffer A (residual proteins II), but was only ex-
tracted in boiling buffer A from protease-treated
cooked rice (Table III). Protease-treated cooked
rice had higher residual protein than the cor-
responding raw rice.
Rat TD correlated positively with the three in
vitro assays, EDN of cooked rice and DP of raw
and cooked rice (Table IV). EDN correlated sig-
nificantly with DP of raw and cooked rice and
NPU and with lysine content of protein and neg-
atively with cysteine content in protein and be-
haved closer to in vitro DP of raw rice than to DP
of cooked rice. In vitro DP of cooked rice correlated
with DP of raw rice and negatively with cysteine
content of protein. In vitro DP of raw rice correlated
with NPU and lysine content of protein and neg-
atively with cysteine content of protein.
Prolamin and other protein fractions
Total protein extraction from raw rice was 94–
106% (mean 100%)(Table III). Albumin-globulin
187
values were low at 0·25–0·40% (4–6% of protein).
Low prolamin values of 0·23–0·44% (3–7% of pro-
tein) were obtained using 60% 1-PrOH solvent
without
-mercaptoethanol. Glutelin values were
high (4·80–6·87% of milled rice; 87–91% of protein)
but included the 10–16 kD prolamin subunits, as
verified by SDS-PAGE. The addition of
-mer-
captoethanol to 60% 1-PrOH should have ex-
tracted the 10–16 kD prolamin subunits. Rat TD
did not correlate significantly with albumin-glob-
ulin and prolamin contents, but correlated neg-
atively with glutelin content (Table IV). Correcting
for the prolamin contamination in glutelin, the cor-
rected prolamin values of raw rice were 0·72–1·22%
(11–17% of protein) and corrected glutelin values
were 3·90–6·09% (72–83% of protein).
Cooking, as expected, reduced protein ex-
tractability to 64–81% (mean 71%) due to heat
denaturation (Table III). ‘Albumin-globulin’ val-
ues were 0·12–0·26% (2–5% of protein), ‘pro-
lamin’ content was 0·05–0·09% and ‘glutelin’
content was 4·16–5·63%.
The denatured prolamin (10–16 kD) values in
cooked rice flour, which was extracted only by
boiling and not by cold buffer A, was 0·13–0·34%
of cooked rice and was lower in Koshihikari,
Sasanishiki, Sinandomeng and Toro 2 than in
PR23383-15, IR24, PRJ 3 and Khao Dawk Mali
105 (Table III). Correlation coefficient with rat
TD was significant (Table IV). The denatured
prolamin probably represents the high cysteine
prolamin fraction29 that became denatured on
cooking due to disulphide bonding to cystine.
Which explains why TD correlated with cysteine
content of protein (Table IV). Protein content of
cooked rice in Table III was lower than that in
Table II, except for Koshihikari. However, the
protein values from the two laboratories were
correlated (r=0·96, p<0·01).
Rat TD correlated positively with lysine content
in the protein and negatively with protein content,
glutelin content and with cysteine and tyrosine
levels in protein (Table IV). Interestingly DIAS
protein content correlated with lysine (r=−0·75,
p<0·05) and tyrosine (r=0·91, p<0·01) in protein
and with glutelin content (r=0·85, p<0·01) and
may explain the significant correlation of TD with
these properties.
Waxy gene product
Rat TD did not correlate significantly negatively
with waxy gene product content of milled rice
188 S. Boisen et al.

Table III Properties of raw and cooked milled rices (Institute of Plant Breeding, UP Los Baños-PhilRice Los Baños)

Property of Koshihikari Sasanishiki Toro 2 Sinandomeng PRJ 3 Khao PR23383-15 IR24

milled rice Dawk
Mali 105

Raw milled rice

Protein (% N×62·5) 5·35 5·46 6·44 5·83 7·38 7·74 7·91 6·64
±SD ±0·06 ±0·25 ±0·42 ±0·06 ±0·16 ±0·16 ±0·07 ±0·05
In vitro digestible 89·2 88·6 87·2 81·4 82·2 82·8 82·7 81·1
protein (%) ±SD ±0·2 ±0·0 ±0·4 ±0·0 ±1·2 ±0·4 ±1·0 ±0·4
Res. proteins I (%) 2·5 1·6 1·5 3·6 1·6 1·8 1·0 1·6
Res. proteins II (%) 9·5 8·6 12·7 9·9 6·1 4·7 6·0 8·0
Albumin-globulina,b (%) 0·34c 0·25f 0·34c 0·27e 0·38b 0·27e 0·31d 0·40a
Prolamina,b (%) 0·30d 0·27e 0·23f 0·23f 0·32c 0·44a 0·34b 0·44a
Glutelina (%) 4·80f 5·18e 5·54a 5·68c 6·77a 6·87a 6·49b 5·39d
±SD ±0·02 ±0·00 ±0·01 ±0·02 ±0·11 ±0·07 ±0·05 ±0·05
Percent N extraction 102 104 95 106 101 98 102 94
Waxy gene product (%)
Total 0·07 0·06 0·11 0·08 0·06 0·08 0·13 0·20
After proteolysis 0·09 0·06 0·10 0·08 0·06 0·08 0·11 0·08
Cooked milled rice
Protein (% N×6·25) 6·10 6·21 7·79 6·91 7·92 8·35 7·51 7·17
±SD ±0·02 ±0·25 ±0·13 ±0·02 ±0·27 ±0·06 ±0·09 ±0·36
In vitro digestible 80·0 81·1 82·6 78·0 77·9 79·1 77·6 77·0
protein (%) ±SD ±0·4 ±0·3 ±0·1 ±0·3 ±1·3 ±0·7 ±0·6 ±0·8
Res. protein II (%) 20·5 20·5 12·7 13·1 16·5 17·5 14·3 16·3
“Albumin-globulin”a,b (%) 0·26a 0·22b 0·19d 0·21c 0·12g 0·15f 0·16e 0·19d
“Prolamin”a,b (%) 0·08b 0·07c 0·07c 0·06d 0·09a 0·06d 0·05e 0·07c
“Glutelin”b (%) 4·56e 4·16f 5·63a 4·20f 4·85c 5·63a 4·75d 5·25b
±SD ±0·01 ±0·00 ±0·02 ±0·00 ±0·07 ±0·03 ±0·06 ±0·03
Percent N extraction 80 72 76 65 64 70 66 77
Denatured “prolamin”
(%) 0·17 0·15 0·13 0·17 0·34 0·32 0·31 0·27
Waxy gene product (%)
Total 0·07 0·06 0·11 0·08 0·06 0·08 0·13 0·20
After proteolysis 0·00 0·00 0·00 0·00 0·00 0·00 0·00 0·00
a
SD<0·01%.
b
Protein values in the same line followed by the same letter are not significantly different at p=0·05 by Duncan’s27 multiple
range test.
c
No residual protein I was extracted.

(Table IV). SDS-PAGE of residual protein of in
vitro protease-digested milled rice showed mainly
the 13±3 kD prolamin band in both raw and
cooked rice, but only the raw rice had some waxy
gene product at MW 60 kD. By contrast, TD of
raw rice protein in rats is 100%2. The enzymes
used probably did not have adequate amylase
activity to hydrolyze the raw starch protecting the
waxy gene product in vitro. However, the decrease
in waxy gene product in IR24 raw rice from 0·20%
to 0·08% due to proteolysis (Table III) is difficult
to explain. The gelatinized starch in cooked rice
facilitated the in vitro hydrolysis of waxy gene
product.
The positive but not significant correlation be-
tween AC and rat TD (Table IV) was because
of the higher AC content of Koshihikari and
Sasanishiki (Table I). This contrasts with the earlier
negative value on 19 cooked milled rices differing
in AC and GT2,6–10. Protein content and AC were
also correlated (r=−0·87, p<0·01).
The poor starch digestibility of IR36 based
 Rice protein digestibility and properties 189

Table IV Simple correlation coefficients between digestibility values and other grain properties of eight
milled rices

Property of milled rice Simple correlation coeeficients with

TD EDN DPcooked DPraw

True digestibility (TD) in rats 1·00 0·93∗∗ 0·83∗ 0·95∗∗

(% of cooked rice protein intake)
Enzyme digestible nitrogen (EDN) 1·00 0·81∗ 0·98∗
(% of cooked rice protein)
Digestible protein of cooked rice 1·00 0·83∗
(DPcooked)(% of protein)
Digestible protein of raw rice 1·00
(DPraw)(% of protein)
Enzyme digestible dry matter 0·63 0·51 0·66 0·48
(% of cooked rice dry matter)
Biological value (% of absorbed N) 0·58 0·50 0·18 0·52
Net protein utilisation (% of 0·80∗ 0·72∗ 0·44 0·74∗
cooked rice ingested N)
Protein content (DIAS) (% in −0·75∗ −0·62 −0·39 −0·71∗
cooked rice)
Protein content (LB) (% in cooked) −0·64 −0·51 −0·17 −0·57
Protein content (LB) (% in raw) −0·76∗ −0·57 −0·39 −0·64
Lysine in protein (g/16 g N) 0·93∗∗ 0·89∗∗ 0·62 0·86∗∗
Cysteine in protein (g/16 g N) −0·83∗ −0·88∗∗ −0·72∗ −0·85∗∗
Tyrosine in protein (g/16 g N) −0·78∗ −0·56 −0·52 −0·65
Albumin-globulin (% in raw rice) −0·34 −0·31 −0·30 −0·23
Prolamin (% in raw rice) −0·57 −0·37 −0·55 −0·47
Glutelin (% in raw rice) −0·71∗ −0·55 −0·41 −0·63
Denatured prolamin (% in cooked −0·82∗ −0·61 −0·72∗ −0·68
rice)
Amylose content (% in raw rice) 0·61 0·61 0·41 0·70
Waxy gene product (% in rice) −0·52 −0·49 −0·40 −0·43
∗ ∗∗
Significant at p=0·05. Highly significant at p=0·01.

amylose extender mutant even in raw form8 is prob-
ably due to the presence, together with the poly-
hedral granules, of irregularly shaped ae granules29,
which waxy gene product is not readily digested
probably because of a different granule structure
and higher GT. By contrast its polyhedral granules
are readily digested with their waxy gene product.
Waxy milled rices
Verification of the above results was undertaken
with two pairs of japonica and indica waxy cooked
milled rices, which are devoid of the waxy gene
product11,12. Glaszmann18 method verified the clas-
sification for the Philippine rices (Table V), but
the Taiwanese rices were received as milled rice
and could not be tested. However, Taikeng Glu.
5 is a short grain typical of japonica rice and
Taichung Sen Glu. 1 is a long grain typical of
indica rice. The indica rice had higher protein
content, particularly for the Taiwanese pair (Table
V).
The waxy samples had similar digestible energy
values (Table V). The indica samples tended to
have lower TD but only the difference for the
Taikeng-Taichung pair was significant. Differ-
ences in BV and NPU were not significant. The
EDN and EDDM values tended to be lower also for
indica rices but the differences were also significant
only for the Taiwanese pair. Amino acid score X
TD was also lowest for Taichung Sen Glu. 1.
Lysine content of protein and amino acid score
were lower in Taichung Sen Glu. 1 than in the
others. Indica rice protein had higher cysteine and
methionine contents than japonica rice protein.
The tyrosine trend in the eight low-AC rices (Table
IV) was not shown by the waxy rices. The lysine
difference probably reflected the difference in pro-
tein content31, but protein content showed a neg-
ative correlation with cysteine in protein and a
190 S. Boisen et al.

Table V Properties, rat balance data, aminogram and in vitro digestibility of two pairs of
japonica and indica cooked waxy milled rices

Property Philippine Taiwanese

Malagkit IR29 Taikeng Taichung

Sungsong Glu. 5 Sen Glu. 1

Glaszmann’s18 group VI I (VI) (I)

Amylose content (% dry basis) 1·7 2·0 1·3 1·8
Gross energy ( J/g dry matter) 17·3 17·2 17·4 17·5
Rat balance data
Digestible energy (% ±SD) 98·0±0·4 96·9±0·7 97·2±0·6 97·0±0·5
True digestibility (% ±SD) 97·9±1·2 93·8±3·0 97·2±1·9 93·1±0·9
Biological value (% ±SD) 77·2±2·2 76·5±1·7 73·7±2·9 71·4±3·3
NPU (% ±SD) 75·5±2·1 71·7±2·1 71·6±3·0 66·5±3·7
EDNa (% at ileal level) 85·1 81·7 87·3 77·0
EDDMb (% at ileal level) 80·5 74·1 80·4 71·1
Protein (% N×6·25 dry basis) 9·19 9·58 9·88 10·75
Albumin-globulin (% of raw) 0·3 0·3 0·4 0·2
Prolamin (% of raw rice) 0·5 0·6 0·8 0·6
Glutelin (% of raw rice) 8·4 8·7 8·7 9·9
Lys (g/16 g N) 3·2 3·2 3·2 3·0
Met (g/16 g N) 2·4 2·7 2·4 2·6
Cys (g/16 g N) 2·1 2·3 2·1 2·3
Thr (g/16 g N) 3·3 3·3 3·2 3·3
Ile (g/16 g N) 4·3 4·4 4·3 4·3
Leu (g/16 g N) 8·0 7·9 7·8 7·7
Val (g/16 g N) 5·9 5·8 5·8 5·8
Phe (g/16 g N) 5·0 4·9 4·8 4·9
Tyr (g/16 g N) 4·0 3·9 4·0 4·1
Arg (g/16 g N) 7·9 7·3 8·0 7·4
His (g/16 g N) 2·4 2·3 2·4 2·3
Ala (g/16 g N) 5·3 5·3 5·2 5·2
Asp (g/16 g N) 8·5 8·1 8·4 8·0
Glu (g/16 g N) 17·4 17·0 16·9 17·0
Gly (g/16 g N) 4·2 4·1 4·3 4·0
Pro (g/16 g N) 4·3 4·4 4·4 4·3
Ser (g/16 g N) 5·2 5·0 5·1 5·0
N recovery (%) 90·7 89·3 89·0 90·4
Amino acid scoreb (%) 54 56 55 52
Amino acid scoreb×TD (%) 53 53 53 48
a
EDN means enzyme digestible protein (N) at ileal level; EDDM means enzyme digestible dry
matter at ileal level.
b
Based on 5·8% lysine in FAO/WHO/UNU22 pattern as 100%.

positive but not significant correlation with tyros-
ine in protein31. The prolamin content was the
highest for Taikeng Glu. 5. Denatured prolamin
in cooked rice could not be estimated because of
the greater solubility of gelatinised waxy starch in
buffer A.
Thus this study of eight low-AC and four waxy
low-GT cooked milled rices verified the higher rat
TD and in vitro protein digestibility of japonica
rice over indica rice. The difference was not related
to lower prolamin content and lower waxy gene
product level of starch in japonica, but was related
to lower cysteine level in protein and lower level
of denatured prolamin in the cooked rice. The
higher level of cysteine in indica protein probably
contributes to this greater prolamin denaturation.
The higher protein content in indica rice was
verified.
Acknowledgements
We would like to thank the suppliers of the rice samples,
the technical assistance of L.T. Roferos, the Bio-
chemistry and Analytical Services Laboratory, Institute
of Plant Breeding, UPLB and the IRRI Isozyme
Laboratory and editing by Tess Rola. J.B.A.D. ac-
knowledges the scholarship extended by PhilRice for
his M.S. programme at UPLB.
 Rice protein digestibility and properties
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