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Journal of Food Protection, Vol. 47, No. 6, Pages 441-444 (June 1984)
Copyright®, International Association of Milk, Food, and Environmental Sanitarians
Germination Effects on Flatus-Causing Factors
and Antinutrients of Mungbeans and Two Strains
of Small-Seeded Soybeans
1,2 1 3
AMINAH ABDULLAH , RUTH E. BALDWIN * and HARRY MINOR Downloaded from http://meridian.allenpress.com/jfp/article-pdf/47/6/441/1656196/0362-028x-47_6_441.pdf by Indonesia user on 04 January 2023
Department of Food Science and Nutrition and Department of Agronomy, University of Missouri-Columbia, Columbia, Missouri 65211
(Received for publication October 13, 1983)
ABSTRACT before they are consumed, but they are important in spr-
outs which receive only a mild heat treatment before they
Two strains of small-seeded soybeans contained greater are eaten.
amounts of stachyose and raffinose than market samples of The interaction of phytic acid (PA) with protein, vita-
mungbean seeds. These sugars either disappeared or were re- mins and several minerals is one of the primary factors
duced to trace amounts during 3 d of germination for mung- limiting the nutritive value of cereal grains and pulse pro-
beans and 4 d for soybeans. Both soybean strains contained teins. Interference of dietary phytate with mineral
more phytic acid and trypsin inhibitor than mungbeans. Germi- metabolism has been well-documented (16,19). If hydro-
nation reduced these antinutrients in both legumes. Phytic acid lyzed, however, PA would be a source of inositol and
(wet weight basis) in one strain of soybean sprouts did not dif- inorganic phosphate. Thus, if the phytate in legume seeds
fer from that in the mungbean sprouts. could be hydrolyzed, their nutritional value would be im-
proved.
Reports of the beneficial effect of germination on the PA is apparently utilized by legumes as a source of
flatus-causing factors of soybeans and other legumes are phosphorus during germination. Increased phytase activity
somewhat contradictory. Reductions ranging from 70 to resulting in an increase in inorganic phosphorus and a
100% in the oligosaccharides, stachyose and raffinose, decrease in PA was reported by Lolas and Markakis (17)
after 3 or 4 d of germination of beans have been reported and Mandal et al. (18). Also, Chen and Pan (6) observed
(1,9,12,23). In contrast, Calloway et al. (4) observed no that phytase activity increased over 200% during the first
reduction in flatus activity in germinated soybeans. The 5 d of germination of soybeans.
wide variance in findings may be due to differences in Much of the available information regarding composi-
type of bean, conditions of germination or activity of en- tional changes during soybean germination is based on
zymes, particularly a-galactosidase. studies of cultivars developed for grain markets. For this
There is also some lack of agreement in results of in- study, two small-seeded soybean [Glycine max (L.) Mer-
vestigations of the effects of germination on the antinutri- rill] strains, SP 75051 and Jaeraejong 320-7, were in-
tional effects of protease inhibitors in soybeans. Rackis troduced from the Crop Experiment Station, Organization
(20) reviewed the pertinent literature and attributed a for Rural Development, Suweon, Korea. In Korea, these
marked pancreatic hypertrophic response in rats to trypsin lines had been selected for their ability to produce high
inhibitors in raw soybean meal. Desikachar and De (7) quality sprouts (personal communication, Y. W. Kim).
reported trypsin inhibitor activity was not lost during ger- The investigation was designed to evaluate the effects of
mination, but Bates et al. (2) stated trypsin inhibitor ac- germination on the flatulence factors (raffinose and
tivity was decreased by two-thirds during germination of stachyose) and antinutrients (PA and trypsin inhibitor ac-
soybeans. Freed and Ryan (//) demonstrated only minor tivity) in these two strains of small-seeded soybeans de-
changes in trypsin inhibitor activity during germination veloped for sprout production. A single strain of mung-
and noted the amount was dependent on the soybean beans was used for comparison, because mungbeans are
strain. Since trypsin inhibitors are thermolabile, they are preferentially used for sprout production in the United
not important in products that receive a heat treatment States.
MATERIALS AND METHODS
'Department of Food Science and Nutrition. SP 75051 and Jaeraejong 320-7 soybean strains were multiplied near
-Present address: Department of Food Science and Nutrition, National Columbia, MO, to obtain adequate quantities of seed for study. A
University of Malaysia, Bangi, Selangor, Malaysia. mungbean strain was purchased from a local market (Park Oriental
^Department of Agronomy. Store, Columbia, MO). The bean seeds were sorted by hand to remove
JOURNAL OF FOOD PROTECTION, VOL. 47, JUNE 1984
442 ABDULLAH, BALDWIN AND MINOR
broken units and extraneous material, and were then stored in a tightly seeds. The seeds of the Jaeraejong 320-7 strain contained
sealed container at room temperature (max. 6 months). Portions of the 0.89% raffinose and 4.87% stachyose, whereas the SP
seeds were prepared for analyses by grinding to pass through a 40-mesh 75051 strain of soybean contained 0.99% raffinose and
screen and kept in tightly sealed plastic bags at 5°C until analyzed. 4.33% stachyose. The mungbean seeds contained 0.45%
Seeds to be germinated were first immersed for 2 min in a 1.6% and 2.19% raffinose and stachyose, respectively (Table
sodium hypochlorite solution to prevent mold growth. Then, they were
soaked in tap water for 8 h. For germination, seeds were held for 3 d 1). Hymowitz and Collins (73) reported considerable var-
at 28°C in a moist atmosphere. iation in sugar content among 195 soybean cultivators.
Analysis of raffinose and stachyose The raffinose content of both Jaeraejong 320-7 and SP
The extraction procedure for gas liquid chromatographic (GLC) anal- 75051 strains of soybeans fell within the range for this
ysis of oligosaccharides was that of Hymowitz et al. (14). One gram sugar, but stachyose exceeded the range found by
of powdered seeds or sprouts (dry weight basis) was used. The derivati- Hymowitz and Collins (13).
zation of sugars was the method of Brobst and Lott (3), with minor Raffinose and stachyose either completely disappeared
modifications. or remained in only trace amounts by the third day of
A Bendix series 2600 chromatograph (Bendix Corp., Lewisburg, germination for mungbeans and the fourth day for soy-Downloaded from http://meridian.allenpress.com/jfp/article-pdf/47/6/441/1656196/0362-028x-47_6_441.pdf by Indonesia user on 04 January 2023
WV) was employed. The stainless steel column was U-shaped, 1.8 m beans. By the third day of germination, the quantities of
in length and 0.64 cm in diameter. It was packed with 3% OV 107
on Chromosorb W (80 to 100 mesh). Before use, the column was con- stachyose had been reduced to levels which did not differ
ditioned for at least 12 h at 300°C with a carrier gas flow rate of 20 significantly (P<0.05) from those of day 4 (Table 1).
ml/min. Once each week, the column was injected with Silyl-8 GLC Prolongation of germination beyond 3 d was deemed in-
column conditioner (10 to 50 jj.g) with the column temperature at advisable because of increased rootlet growth and in-
150°C. To ensure efficient resolution and reproducibility, the oven tem- creased probability of mold growth. In general, trends
perature was maintained continuously at 300°C when the chromatograph
was not in operation. The septum was changed daily and the detector were similar to those reported by Adjei-Twum (1), East
inlet was cleaned as needed. et al. (9) and Hsu et al. (12). Variation in the number
Chromatograms were recorded by a Honeywell Electronic 194 two- of days of germination required for complete removal of
pen 10-in. laboratory recorder (Honeywell Industrial Division, Ft. stachyose and raffinose was attributed by Hymowitz et
Washington, PA). The areas of the peaks were integrated manually al. (14) to varietal differences. Cartter and Hopper (5)
using a computing integrator (Infotronics Model CRS-309, International indicated variation may also be due to the location in
Tech. Instruments, Boulder, CO).
which the seeds were grown. According to Reddy and
Analysis of phytic acid (PA) and trypsin inhibitor activity Salunkhe (22), the disappearance of oligosaccharides dur-
The PA in bean seeds and sprouts was determined by ferric chloride ing germination is caused by enzymatic degradation, as
precipitation from an extract as described by Early (§). The amount confirmed by increased a-galactoside activity in germi-
of phosphorus (phytate phosphorus) in the extract was determined using nated seeds. Tada and Kawamura (25) observed the de-
the Fiske and Subbarow (10) procedure. crease in oligosaccharides during germination was fol-
Trypsin inhibitor activity was determined according to the method
of Kakade et al. (15). This is a colorimetric procedure with absorbance lowed by an increase in reducing sugars.
measured at 410 nm. An increase of 0.01 absorbance unit represents
one trypsin unit. Trypsin inhibitor activity is expressed as trypsin in- Phytic acid (PA)
hibitor units per mg (TlU/mg). On both a wet and a dry weight basis, the seeds of
RESULTS AND DISCUSSION the two soybean strains were significantly higher in PA
content than mungbean seeds or sprouts. Germination re-
In contrast to the 12 to 18 g/100 seeds considered nor- sulted in a significant (P<0.05) reduction in PA in both
mal for soybeans, SP 75051 and Jaeraejong 320-7 had soybeans and mungbeans on a wet weight basis. Because
mean 100-seed weights of 7.7 g and 9.1 g, respectively. of the large reduction of PA in Jaeraejong 320-7, the PA
The mungbeans averaged 5.9 g per 100 seeds. in sprouts of this variety did not differ from that of
mungbean sprouts on a wet weight basis. The reduction
Flatulence factors: Raffinose and stachyose of PA was approximately four-, three-, and four-fold for
The soybean seeds contained greater amounts (% dry soybean Jaeraejong 320-7, SP 75051, and mungbean, re-
weight basis) of raffinose and stachyose than mungbean spectively, on a wet weight basis. On a dry weight basis,
TABLE 1. Mean" percent (dry weight basis) rqffinose and stachyose of soybean and mungbean seeds during germination.
Raffinose Stachyose
Soybean Mungbean Soybean Mungbean
Germination Jaeraejong Jaeraejong
period 320-7 SP 75051 320-7 SP 75051
Seed 0.89A 0.99A 0.45A 4.87 A 4.33A 2.19A
Day 1 0.75B 0.88B 0.39A 2.92B 3.17B 2.18A
Day 2 0.45C 0.32C 0.06B 1.71C 1.05C 0.06B
Day 3 0.19D 0.14D 0.02B 0.37D 0.29D 0.01B
Day 4 0.01E 0.00E 0.00B 0.00D 0.00D 0.00B
aN = 4. Where letters differ within a column, means differ significantly (P<0.05) from each other.
JOURNAL OF FOOD PROTECTION. VOL. 47, JUNE 1984
GERMINATION EFFECTS ON MUNGBEANS AND SOYBEANS 443
there was a reduction in PA content, but it was statisti- REFERENCES
cally significant (P<0.05) only for the Jaeraejong 320-7
soybean (Table 2). 1. Adjei-Twum, D. C. 1976. Studies on the germination, growth, and
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TABLE 2. Mean" phytic acid content and trypsin inhibitor activity in soybean and mungbean seeds and sprouts.
Phytic acid Trypsin inhibitor activity
Legume and
germination state Wet weight Dry weight Wet weight Dry weight
(mg/g) (mg/g) (TlU/mg) (TlU/mg)
Soybean
Jaeraejong 320-7 seed 8.10A 8.72A 31.85A 34.32A
Jaeraejong 320-7 sprout 1.50DE 6.02B 6.75B 27.66B
SP 75051 seed 6.93B 7.44AB 30.25A 32.46A
SP 75051 sprout 1.63D 6.64B 5.85B 25.11B
Mungbean seed 2.83C 3.12C 5.41B 5.96C
Mungbean sprout 0.53E 2.52C 0.80C 3.88C
a
N = 5. Where letters differ within a column, means differ significantly (P<0.05) from each other.
JOURNAL OF FOOD PROTECTION, VOL. 47, JUNE 1984
444 ABDULLAH, BALDWIN AND MINOR
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JOURNAL OF FOOD PROTECTION. VOL. 47, JUNE 1984
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