Journal of Food and Nutrition Research
ISSN (Print): 2333-1119 ISSN (Online): 2333-1240 Website: Editor-in-chief: Prabhat Kumar Mandal
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Journal of Food and Nutrition Research. 2020, 8(10), 561-567
DOI: 10.12691/jfnr-8-10-4
Open AccessArticle

Optimizing of Enzyme Hydrolysis Condition for Bitterness Suppression of Soybean Protein Using Response Surface Methodology (RSM)

Ha-Na Ra1, Yong-Sik Cho1, Young Hwang1, Hyun-Wook Jang1 and Kyung-Mi Kim1,

1Fermented & Processed Food Science Division, Department of Agrofood Resources, National Institute of Agricultural Sciences, Rural Development Administration, Jeollabuk-do, Republic of Korea

Pub. Date: October 22, 2020

Cite this paper:
Ha-Na Ra, Yong-Sik Cho, Young Hwang, Hyun-Wook Jang and Kyung-Mi Kim. Optimizing of Enzyme Hydrolysis Condition for Bitterness Suppression of Soybean Protein Using Response Surface Methodology (RSM). Journal of Food and Nutrition Research. 2020; 8(10):561-567. doi: 10.12691/jfnr-8-10-4


Enzyme hydrolysis of soybean protein can be adjusted according to purposes, but should be optimized for reaction time, concentration and substrate conditions to prevent bitterness caused by excessive hydrolysis. This study was carried out to find the optimal condition of the soybean protein hydrolysis process using a response surface methodology. The experiment was designed based on a central composite design, and the independent variables were the soybean protein extraction pH (X1, 10-12), enzyme concentration (X2, 0.1-0.5%) and hydrolysis time (X3, 150-210 minutes). The results of the degree of hydrolysis (Y1), pH (Y2) and soluble solid contents (Y3) were fitted to a response surface methodology model (R2= 0.91, 0.98, and 0.86, respectively). The optimal hydrolysis condition for soybean protein hydrolysis was as follows; pH 12, enzyme concentration 0.27% and 187.88 min, respectively. While richness and sourness were increased with soybean protein hydrolysis, the saltiness was decreased. The bitterness of the hydrolysate prepared at the optimal condition did not show any difference compared with that of the soybean protein, whereas richness and sourness showed a significant increase. The enzyme hydrolysate at the optimal condition showed distribution of a molecular weight lower than the soybean protein did to form intense bands at 35, and 25 kDa. Therefore, this hydrolysate was expected to be used as a high-value plant protein food material.

enzyme hydrolysis soybean protein optimization RSM

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[1]  Spada, J.C., Marczak, L.D.F., Tessaro, I.C. and Cardozo, N.S.M, “Interactions between soy protein from water-soluble soy extract and polysaccharides in solutions with polydextrose,” Carbohydrate Polymers, 134 (10). 119-127. Dec.2015.
[2]  Zhou, Y., Li, X., Hua, Y., Kong, X.Z., Zhang, C.M., Chen, Y.M. and Wang, S.D, “The absence of lipoxygenase and 7S globulin of soybeans and heating temperatures on the properties of soymilks and soy yogurts,” LWT, 115 (1). 108431. Nov.2019.
[3]  McCann, T.H., Guyon, L., Fischer, P. and Day, L, “Rheological properties and microstructure of soy-whey protein,” Food Hydrocolloids, 82 (9). 434-441. Sept.2018.
[4]  Li, D., Li, X., Wu, G., Li, P.Y., Zhang, H., Qi, X.G. Wang, L., and Qian, H.F, “The characterization and stability of the soy protein isolate/1-Octacosanol nanocomplex,” Food Chemistry, 297 (1). 124766. Nov.2019.
[5]  Jose, J., Pouvreau, L. and Martin, A.H, “Mixing whey and soy proteins: Consequences for the gel mechanical response and water holding,” Food Hydrocolloids, 60 (10). 216-224. Oct.2016.
[6]  Marengo, M., Akoto, H.F., Zanoletti, M., Carpen, A., Buratti, S., Benedetti, S., Barbiroli, A., Johnson, P.N.T., Dawson, E.O.S., Saalia, F.K., Bonomi, F., Pagani, M.A., Manful, J. and Iametti, S, “Soybean-enriched snacks based on Arfican rice,” Foods, 5 (2). 38-48. May.2016.
[7]  Kim J.S. and Shin, J.K, “Sensory characteristics of enzymatically hydrolyzed isolated soy protein by descriptive analysis,” Food Engineering Progress, 23 (1). 39-46. Feb.2019.
[8]  Jeon, S.Y., Lee, Y.M., Kim, S.S. and Kim, K.O, “Effect of added hydrolyzed vegetable proteins on consumers’ response for Doenjang (Korean traditional fermented soybean paste) soup,” Food Science and Biotechnology, 29 (1). 45-53. Aug.2020.
[9]  Cho, Y.J. and Chun, S.S, “Effect of phytase, protease and the mixed enzyme of phytase and protease on the extraction and properteis of the protein from abolished soybean meal,” Journal of the Korean Society of Food Science and Nutrition, 29 (1). 57-63. Feb.2000.
[10]  Puppo, C., Chapleau, N., Speroni, F., Anton, M.L., Michel, F., Anon, C. and Anton, M, “Physicochemical modifications of high-pressure-treated soybean protein isolates,” Journal of Agricultural and Food Chemistry, 52 (6). 1564-1571. Mar.2004.
[11]  Moulton, K.J. and Wang, L.C, “A pilot-plant study of continuous ultrasonic extraction of soybean protein,” Journal of Food Science, 47 (4). 1127-1129. Jul.1982.
[12]  Ra, H.N., Park, S.R., Kim, H.Y., Cho, Y.S. and Kim, K.M, “Optimization of the high-pressure condition for rice protein extracting using response surface methodology (RSM)” Journal of the Korean Society of Food Culture, 34 (6). 779-784. Dec.2019.
[13]  Liu, C., Wang, H., Cui, Z., He, X., Wang, X., Zeng, X. and Ma, H, “Optimization of extraction and isolation for 11S and 7S globulins of soybean seed storage protein,” Food Chemistry, 102 (4). 1310-1316. Jul.2007.
[14]  Jiang, J., Chen, Y.J. and Xiong, L, “Structural and emulsifying properties of soy protein isolate subjected to acid and alkaline pH-shifting processes,” Journal of Agricultural and Food Chemistry, 57 (16). 7576-7583. Jul.2009.
[15]  Zheng, Z., Li, J., Li, J., Sun, H. and Liu, Y.F, “Physicochemical and antioxidative characteristics of black bean protein hydrolysates obtained from different enzymes,” Food Hydrocolloids, 97 (12). 105222. Dec.2019.
[16]  Singh, T.P., Siddiqi, R. A. and Sogi, D.S, “Statistical optimization of enzymatic hydrolysis of rice bran protein concentrate for enhanced hydrolysate production by papain,” LWT, 99 (1). 77-83. Jan.2019.
[17]  Zang, X., Yue, C., Wang, Y., Shao, M. and Yu, G.P, “Effect of limited enzymatic hydrolysis on the structure and emulsifying properties of rice bran protein,” Journal of Cereal Science, 85 (1). 168-174. Jan.2019.
[18]  Fernández A. and Kelly, P, “pH-stat vs. free-fall pH techniques in the enzymatic hydrolysis of whey proteins,” Food Chemistry, 199 (1). 409-415. May.2016.
[19]  Fu, Y., Liu, J., Hansen, E.T., Bredie, W.L.P. and Lanetsch, R, “Structural characteristics of low bitter and high umami protein hydrolysates prepared from bovine muscle and porcine plasma,” Food Chemistry, 257 (1). 163-171. Aug.2018.
[20]  Maehashi, K. and Huang, L, “Bitter peptides and bitter taste receptors,” Cellular and Molecular Life Sciences, 66 (10). 1661-1671. Jan.2009.
[21]  Kim, M.R. “Bitterness and solubility of soy protein, casein, gluten, and gelatin hydrolysates treated with various enzymes,” Journal of the Korean Society of Food Science and Nutrition, 39 (4). 587-594. Apr.2010.
[22]  Adler-Nissen, J. “Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid,” Journal of Agricultural and Food Chemistry, 27 (6). 1256-1262. Nov.1979.
[23]  Lee, M.K., Kim, J.K. and Lee, S.Y, “Effects of fermentation on SDS-PAGE patterns, total peptide, isoflavone contents and antioxidant activity of freeze-thawed tofu fermented with Bacillus subtilis,” Food Chemistry, 249 (1). 60-65. May.2018.