Radiation Processing And Functional Properties Of Soybean (Glycine Max)
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Radiation Physics and Chemistry 79 (2010) 490–494
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Radiation Physics and Chemistry
journal homepage: www.elsevier.com/locate/radphyschem
Radiation processing and functional properties of soybean (Glycine max)
Mrinal Pednekar a,n, Amit K. Das b, Rajalakshmi Va, Arun Sharma a
Food Technology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085, Maharashtra, India
Department of Food Engineering, CFTRI, Mysore 570020, Karnataka, India
a r t i c l e in f o a b s t r a c t
Article history: Effect of radiation processing (10, 20 and 30 kGy) on soybean for better utilization was studied.
Received 25 June 2009 Radiation processing reduced the cooking time of soybean and increased the oil absorption capacity of
Accepted 28 October 2009 soy ﬂour without affecting its proximate composition. Irradiation improved the functional properties
like solubility, emulsiﬁcation activity and foam stability of soybean protein isolate. The value addition
Keywords: effect of radiation processing has been discussed for the products (soy milk, tofu and tofu fortiﬁed
Radiation processing patties) prepared from soybean.
Soybean & 2009 Published by Elsevier Ltd.
1. Introduction Radiation processing is an ecofriendly technology utilized for
nutritional safety and security. It can be used for value addition
In most developing countries, per capita consumption of such as elimination of ﬂatulence factors (Machaiah et al., 1999;
protein is below the recommended level resulting in widespread Machaiah and Pednekar, 2002), increase in starch and protein
protein calorie malnutrition. Soybeans with 40% protein and 20% depolymerization (Sharif and Farkas, 1993; Nene et al., 1975) and
oil have a great potential of solving the problem of protein calorie extractability.
malnutrition. Soy protein efﬁciently supplements cereal grain We have conducted studies to evaluate the effects of radiation
protein, because it corrects the lysine deﬁciency of cereals. In processing on functional properties like cooking time and the
some cases, for example in corn, it also corrects tryptophan yield of soy milk and tofu. Potato patties prepared from tofu
deﬁciency. Because of its quality, soybean protein can replace incorporation indicated good acceptability. Functional properties
animal protein without a signiﬁcant decrease in nutritive value like solubility, foaming, emulsiﬁcation capacity and gelling of
(Tripathi and Misra, 2005). In fact, soy protein has an advantage protein extracted from soybean were also evaluated.
over animal protein as it does not raise the serum cholesterol
values (Fukushima, 2001) and hence, is useful for people suffering
from cardiovascular disorder (De Kleijn et al., 2002). Soybean also 2. Materials and Methods
has a number of phytochemicals found to be effective in ﬁghting
osteoporosis (Anderson and Garner, 1997), obesity, cancer 2.1. Irradiation
(Messina, 1999) and postmenopausal problems (Albertazzi et al.,
1998). Soy milk is used in cases of lactose intolerance. However, Soybean was purchased from a local market, cleaned and
soybean has a long cooking time, beany ﬂavor, antinutritional irradiated at 10, 20 and 30 kGy. Irradiation was carried out in a
factors like rafﬁnose family oligosaccharides and protease Gamma Cell-5000 loaded with Co60(Board of Radiation and Isotope,
inhibitors (Liener, 1994). Hence, processing of soybean is essential Mumbai, India) at an effective dose rate of 152.3 Gy/min. Dose rate
for better utilization. Most native proteins do not show desirable was determined using standard Fricke dosimetry (Sehsted, 1970).
functional properties and modiﬁcations for improving the nutri- Calibration was done by keeping the dosimeter vials in the
tional value and/or functional properties like protein solubility, irradiation chamber at different positions. Dosimeters were
foaming and gelling need to be induced (Chove et al., 2001). Such analyzed using a UV spectrophotometer. 9% variation in the dose
modiﬁcations imply changes in both protein structure and distribution was recorded.
conformation at different levels by changing molecular composi-
tion or size. 2.2. Soybean ﬂour
Corresponding author. Tel.: + 91 22 25595375. Whole soybean seeds were pulverized into ﬁne ﬂour (710 mm)
E-mail address: email@example.com (M. Pednekar). with a mixer grinder and stored in self sealable polyethylene bags.
0969-806X/$ - see front matter & 2009 Published by Elsevier Ltd.
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2.3. Proximate analysis different sections of the Food Technology Division, BARC. The
panelists belonged to age group of 25–55. The hedonic scale used
Proximate analysis was carried out by standard AOAC methods was as follows:
(AOAC, 2007) for all samples including control after storing at 7=like very much (LVM), 6= like moderately (LM), 5 =like
room temperature for 15 days. Lipid content of soy milk was slightly (LS),
determined by the Folch method (Folch et al., 1957). 4=neither like nor dislike (NLND), 3= dislike slightly (DS),
2= dislike moderately (DM),
2.4. Determination of water and oil absorption capacity 1=dislike very much (DVM).
For water and oil absorption capacities of soy ﬂour the method 2.10. Preparation of soy protein isolate (SPI)
of Sathe et al., (1982) was followed. Soy ﬂour (1 g) was mixed
with 10 mL of distilled water or 10 mL of edible oil and vortexed. Soybean protein isolate was prepared by the isoelectric point
The samples were then allowed to stand at room temperature precipitation method. A modiﬁed method of Sathe et al. (1982) was
(21 1C) for 30 min, and centrifuged at 5000 rpm for 30 min. used to extract protein from the defatted soybean. Finely ground soy
The volume of the supernatant was measured in a 10 mL ﬂour was defatted with petroleum ether (1:20, W/V). Defatted
graduated cylinder. soy ﬂour was treated with 0.2% NaOH for 24 h (1:5, W/V). After
Calculation: centrifuging (10,000 rpm, 30 min), the residue was re-extracted
Water Absorption Capacity (g/g)=(Initial volume of the waterÀ with 0.2% NaOH and again centrifuged. Protein from both the pooled
Final volume of water) supernatants was precipitated by adjusting the pH to 4 with 1 N HCl.
Oil Absorption Capacity (g/g)=(Initial volume of the oil–Final The precipitate was dissolved in a minimum amount of 0.2% NaOH.
volume of oil) Â 0.91n This protein solution was dialyzed against distilled water for 72 h
(n = speciﬁc gravity of the oil used) and freeze-dried.
2.5. Cooking time 2.11. Protein solubility
Overnight soaked seeds (control and irradiated) were pressure Protein solubility of SPI was studied from pH 1–12. The sample
cooked for 2, 4, 6, 8 and 10 min (after ﬁrst pressure release).These (100 mg) was dissolved in distilled water and pH was adjusted to
seeds were then allowed to cool at room temperature. The degree the required value using 0.1 N HCl or NaOH. The volume was
of softness of the seeds was then measured using Texture made up to 20 mL. The protein solution was centrifuged at
Analyzer (TA.XT Plus, Stable Micro System, Surrey, U.K.). 8000 rpm for 15 min. The protein in the supernatant was
estimated using the Kjeldahl method.
2.6. Preparation of soy milk
2.12. Determination of gelling capacity
Soybean seeds (100 g) were soaked for 16 h in 300 mL water.
Gelling was studied using a modiﬁed method of Sathe et al.
Excess water was decanted off and 80 mL fresh water was added
(1982). SPI was dissolved in different concentrations of 10%, 12%,
and seeds were pressure cooked for 10 min after ﬁrst pressure
14% and 16% in phosphate buffer (pH 7.6) and 20% sugar solution.
release. The cooked seeds were blended in the grinder by adding
3 mL of the solution was dispensed in tubes and heated at 80 1C
300 mL of water. Further this homogenate was ﬁltered through
in a water bath for 30 min. The tubes were cooled rapidly by
muslin cloth, while ﬁltering 600 mL of water was added. The
keeping in cold water and kept refrigerated overnight. Gel
ﬁltrate was heated in a boiling water bath for 15 min.
formation was observed by tilting the tubes and observing the
ﬂow of the solution.
2.7. Preparation of tofu
2.13. Foaming properties
Tofu was prepared by a modiﬁed method of Nong Sun and
Breene (1991). Calcium sulfate (0.2 M) was added to warm soy
Whip ability and foam stability were studied according to the
milk in 1:10 ratio and was heated at 70 1C for 10 min for
method of Coffman and Garcia (1977) with slight modiﬁcation. SPI
coagulation to occur. The coagulated curd was separated using
(500 mg) was solubilized in 25 mL distilled water and pH was
cheese cloth and the collected mass (tofu) was weighed.
adjusted to 7.0. The solution was whipped in a homogenizer
(Polytron PT 2100, Kinematica, Switzerland) for 3 min at 10,000 rpm
2.8. Preparation of tofu–potato patties and poured into a 50 mL measuring cylinder. The total and drainage
volume were noted at 0, 1, 30 min, 1, 2, and 24 h intervals.
Potatoes (500 g) were cooked in a pressure cooker and mashed Whip ability and foam stability were calculated by the
with pre-moistened bread slices (4 slices each weighing 22 g). following formula:
This preparation (100 g) was mixed with tofu (15 g). This was Whip ability= (Total volume–Drainage volume)/Initial volume
molded desirably and a layer of semolina was applied to the Foam stability= (Initial volume–Drainage volume)/Initial
surface. This was then shallow fried with vegetable oil to a golden volume Â 100
2.9. Sensory evaluation of tofu–potato patties
Emulsiﬁcation was carried out according to the method of
Sensory evaluation of the tofu–potato patties was carried out Bandyopadhyay and Ghosh (2002). 3 mL of 0.2% SPI solution at pH
using a 7-point hedonic scale ranging from ‘like very much’ to 7 was homogenized with 1, 2 and 3 mL of peanut oil at
‘dislike very much’, with ‘neither like nor dislike’ as the midpoint 12,000 rpm. 100 ml of emulsion was added to 4900 ml of 0.1%
(ASTM, 1996). The taste panel consisted of 50 members from SDS and absorbance was read at 500 nm.
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492 M. Pednekar et al. / Radiation Physics and Chemistry 79 (2010) 490–494
Proximate qualities of soybean.
Sample Protein % Lipid % Moisture% Ash % Carbohydrates%
Control 36.5357 1.704 22.7007 0.582 7.355 7 0.077 0.787 70.033 32.6237 0.293
10 kGy 37.590 7 1.730 22.22570.523 7.345 7 0.190 0.827 70.009 32.013 70.489
20 kGy 36.103 7 1.595 21.6007 1.015 6.475 7 2.27 0.826 70.139 34.9967 0.528
30 kGy 36.65 7 0.826 23.0507 0.825 7.22 70.155 0.733 70.094 32.3477 0.386
The values presented are mean 7S.D. computed from 6 replicates.
2.15. Statistical analysis Table 2
Texturo-metric analysis of cooked soybean.
All values are expressed as means and standard deviation of 6 Force (Kg)a
replicates with the exception of sensory evaluation. The results
are the mean and standard deviations of the observed values. Control 10 kGy 20 kGy 30 kGy
Differences were considered signiﬁcant at po0.05 after perform-
2 min 6.047 7 1.252 3.834 7 0.581 2.4487 1.282 2.505 7 0.781
ing students’t’ test.
4 min 6.646 7 1.923 3.3327 0.960 3.1977 0.964 2.038 7 0.364
6 min 7.140 7 0.764 3.323 7 1.538 3.6527 0.977 1.3117 0.551
8 min 4.581 7 0.942 1.7197 0.726 2.7617 0.133 1.5247 0.658
10 min 4.064 7 0.526 2.028 7 0.987 2.613 7 1.174 1.1577 0.748
3. Results and discussion
S.D. computed from 6 replicates.
3.1. Proximate analysis of soy ﬂour
Proximate analysis of control and irradiated (10, 20 and
3.4. Qualitative analysis of soy milk
30 kGy) soy ﬂour was carried out and the results obtained are
presented in Table 1. It is evident from the table that there was no
It was evident from the data that the protein concentration in
signiﬁcant effect of radiation processing on the macronutrient
soy milk from irradiated seeds nearly doubled (0.58270.08 in
content of the soybean.
control soy milk to 1.04470.1 in 30 kGy treated soy milk).
This can be correlated to better extractability of proteins
3.2. Oil and water absorption capacity from irradiated seeds. Similar results were reported by Byun
and Kang (1994).
Oil and water absorption capacity of ﬂours prepared from
control and radiation processed seeds was found to be within the 3.5. Preparation of tofu
range observed with various legume ﬂours (Adebowale and
Lawal, 2004). Variation in the presence of polar and non-polar The results indicated that irradiation increased the yield of
side chains among ﬂour which bind to water or hydrocarbon side tofu from 5.27% in control to 11.36% in 30 kGy treated samples.
chains of oil possibly make the difference in water or oil-binding The yield increased as the dose increased. This can be correlated
capacity of ﬂours. The data showed a non signiﬁcant increase to the higher concentration of protein in the milk from irradiated
in water absorption capacity due to radiation processing samples. Byun et al., (1993) have observed a similar pattern.
(2.5770.082 in control to 3.0870.075 in 30 kGy treated
samples). Water absorption capacity is important functional
3.6. Sensory evaluation
property for ﬂours as they swell and impart characteristics like
body thickness and viscosity. Radiation processing also led to
The data of the sensory analysis was analyzed and responses
increase in oil absorption capacity of soy ﬂours (1.587 0.16 in
(%) were plotted against hedonic ratings. As seen in Fig. 1, tofu–
control to 1.97 70.18 in 30 kGy samples). Rahma and Mustafa
potato patties, a tertiary product from soybean irradiated at
(1988) have reported similar observations with irradiation
20 kGy dose has scored maximum (40%) as ‘Like very much’,
of peanut ﬂours. Dissociation and denaturation results in the fat
whereas 55% of the panel members rated the same as ‘Like
and water absorption of treated proteins compared to native
moderately’. It is also noteworthy that products from soybean
proteins (Siddharaju et al., 2002). Radiation processing may
irradiated with other doses (10 and 30 kGy) scored more than the
have resulted in protein unfolding leading to exposure of certain
product prepared from control tofu which showed the lowest
buried functional groups resulting in increased oil absorption
scores among all. This can be attributed to the fact that soybean
has a typical beany ﬂavor and taste which are also expected to be
present in its secondary (control tofu) as well as tertiary (tofu–
3.3. Determination of cooking time of soybean potato patties) products. Irradiation may have caused reduction of
the beany ﬂavor, as the scores for the products prepared from
Cooking time is a signiﬁcant characteristic of legumes. It is the irradiated soybean were always higher than those for the control
time of boiling during which the legumes attain desirable softness soybean.
wherein at least 90% of the seeds are soft enough to masticate.
Our results indicated a dose-dependent decrease in cooking time 3.7. Functional properties of soy protein isolate
of irradiated beans (Table 2). There was 30% decrease in cooking
time at 10 kGy which decreased to 60% at 30 kGy. The observed 3.7.1. Protein solubility
reduction in cooking time of irradiated soybean is consistent with The solubility proﬁles of SPI in water at different pH values are
reports of other workers (Rao and Vakil, 1985; Byun et al., 1993). presented in Fig. 2. At pH 4 and 5, which encompasses the
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Product A Product D 700 Control
Product B Product E
100 500 20kGy
0min 5min 0min 5min 0min 5min
1 ml oil 2 ml oil 3 ml oil
Fig. 3. Emulsifying activity index (EAI) of SPI.
Attributes hydrophilic groups which increase the interactions of
hydrophilic amino acids with water molecules. Protein from the
Fig. 1. Sensory evaluation of Tofu fortiﬁed potato patties. A–Control Product 20 kGy treated sample showed a slightly different pattern. It
(without Tofu); B–Control Tofu (mixed with Tofu obtained from un irradiated
Soya); C–Mixed with Tofu obtained from Soybean irradiated with 10 kGy dose;
showed the least solubility up to pH 3, but then increased after
D–Mixed with Tofu obtained from Soybean irradiated with 20 kGy dose; E–Mixed isoelectric point showing higher solubility than the control in the
with Tofu obtained from Soybean irradiated with 30 kGy dose. alkaline range. Physical treatments such as high pressure have
been found to unfold the protein resulting in exposure of
hydrophobic sites altering the functional properties of proteins
(Molina et al., 2002). Irradiation at high doses has been found to
100 alter protein structure. Such alterations must have changed the
protein solubility pattern at different doses due to alterations in
amino acids at the surface as well as length of polypeptides
(Bautista et al., 2006).
In order to form gels, partial denaturation is desirable, since
unfolding of the tertiary structure gives long chains without
breakage of covalent bonds. At the lowest concentration tried
(12%), proper gels could not be obtained with control protein
40 while SPI from irradiated soybean showed gelling at 12%
10 kGy concentration. It is a known fact that factors like pH, ionic
20 kGy strength, reducing agents and the presence of non-protein
20 30 kGy compounds (carbohydrates) affect gelling. Sucrose is a main
ingredient in commercial gel mixes and when gelling was done in
20% sugar solution, even the control sample showed gelling at 12%
0 2 4 6 8 10 12 concentration. The gels were of coagulant type indicating the
pH presence of more non-polar residues (Moure et al., 2006).
Moreover the gels from treated samples were observed to be
Fig. 2. Protein solubility of SPI. homogenous and smooth, when compared with control gels.
isoelectric point of soy proteins, nitrogen solubility was 3.7.3. Foaming properties
signiﬁcantly low for all the samples. This is in concurrence with There was no signiﬁcant difference observed between control
the data obtained by McWatters and Holmes (1979) for soy ﬂour. and irradiated sample at the concentration tried (2%). It is a
As the pH increased above 5, nitrogen solubility increased steeply known fact that ﬂexible protein molecules can have good
reaching almost a plateau around neutral pH. The solubility foamability by reducing the surface tension.
remained consistently higher in the case of protein extracted from
irradiated soybean. SPI extracted from 10 kGy treated samples 3.7.4. Emulsiﬁcation
showed higher solubility (100%) at most of the pH values. It can be seen from Fig. 3 that the emulsiﬁcation activity as
However, in the range encompassing the isoelectric point dose well as the stability of protein extracted from treated seeds was
dependency was evident. The amino-acid composition, signiﬁcantly higher compared to the control. Qi et al., (1997)
particularly at the protein surface inﬂuences protein solubility. observed an increased emulsiﬁcation activity index (EAI) after
Higher solubility is related to the presence of low numbers of pancreatic hydrolysis of soy protein isolate, reaching maximum at
hydrophobic residues (Moure et al., 2006). Irradiation at this dose 15% degree of hydrolysis (DH). It indicates that at 15% DH, the
must have led to formation of smaller peptides exposing hydrophilic and hydrophobic groups are well balanced. We
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494 M. Pednekar et al. / Radiation Physics and Chemistry 79 (2010) 490–494
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