Food and Fermentation Industries >

2023 , Vol. 49 >Issue 22: 102 - 109

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035225

Modifications of soy 7S globulin using enzymatic hydrolysis based on in silico analysis

  • CHEN Lin ,
  • ETTELAIE Rammile ,
  • YU Lin ,
  • CHEN Wei
Expand
  • 1(Guangdong Provincial Key Laboratory of Plant Resources Biorefinery, School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510006, China)
    2(School of Food Science and Nutrition, University of Leeds, Leeds LS29JT, United Kingdom)
    3(The Affiliated Huadu Hospital of Southern Medical University, Guangzhou 510800, China)

Received date: 2023-02-21

  Revised date: 2023-03-09

  Online published: 2023-12-25

Fold

Abstract

To efficiently select proteases appropriate for improving the protein solubility of soy 7S globulin under acidic conditions, 7S was proteolyzed in silico using the software PeptideCutter, and then the isoelectric points (pI) of produced peptides were predicted using the on-line software Compute pI/Mw. Peptides with pI>pH 6.0 or pI<pH 3.0 were selected to construct peptide library, whereby the appropriate proteases were screened and were verified through in vitro experiments. Results showed that proteases that could produce 20 or more target peptides from in silico analysis were protease K, pepsin, thermolysin, chymotrypsin, and trypsin in sequence. Results of in vitro experiments verified that these five proteases could improve the protein solubility (PS) of 7S at pH 3.0-4.5 to varying degrees, with protease K performing the best. The PS of 7S hydrolysates modified by protease K was approximately 81% at pH 3.0-5.0. Analysis of electrophoresis and Zeta potential of 7S hydrolysates demonstrated that the α′, α and β subunits of 7S underwent total degradation after proteolysis using the five advantageous proteases, and the resultant 7S hydrolysates mainly consisted of peptides with molecular weights<3 000 Da, and showed a shift in pI to acidic conditions. These findings were consistent with those of in silico analysis. However, due to the complexity of proteolysis of 7S, the in silico analysis was insufficiently accurate when the quantities of target peptides produced by different proteases were not much different. Stability analyzer results showed that 7S hydrolysate modified by protease K was more stable against flocculation and precipitation than the control 7S in orange juice. This study demonstrated that based on in silico analysis, the screening efficiency of appropriate proteases for enzymatic modifications of 7S could be increased markedly, and the resultant 7S hydrolysates showed good protein solubility under acidic conditions and have a potential application prospect in acidic beverages.

Key words： soy 7S globulin; in silico analysis; enzymatic modification; protein solubility; acidic beverage

Cite this article

CHEN Lin , ETTELAIE Rammile , YU Lin , CHEN Wei . Modifications of soy 7S globulin using enzymatic hydrolysis based on in silico analysis[J]. Food and Fermentation Industries, 2023 , 49(22) : 102 -109 . DOI: 10.13995/j.cnki.11-1802/ts.035225

References

[1] 李佳笑, 石爱民, 刘红芝, 等.植物蛋白酸性条件下溶解性提高的改性方法及应用研究进展[J].中国油脂, 2019, 44(9):59-65.
LI J X, SHI A M, LIU H Z, et al.Progress in modification methods to improve plant protein solubility under acidic conditions and its application[J].China Oils and Fats, 2019, 44(9):59-65.
[2] 黄橙子. 酸溶性大豆蛋白的制备及其在酸性饮料中的应用[D].上海:华东师范大学, 2013.
HUANG C Z.Preparation of acid soluble soybean protein and its application in acidic drinks[D].Shanghai:East China Normal University, 2013.
[3] NISHINARI K, FANG Y, GUO S, et al.Soy proteins:A review on composition, aggregation and emulsification[J].Food Hydrocolloids, 2014, 39: 301-318.
[4] 孙雪, 赵晓燕, 朱运平, 等.酶解大豆分离蛋白的功能性及应用研究进展[J].粮食与油脂, 2021, 34(9):14-17.
SUN Y, ZHAO X Y, ZHU Y P, et al.Research progress on function and application of enzymatic hydrolysis of soy protein isolate[J].Cereals & Oils, 2021, 34(9):14-17.
[5] TAVANO O L.Protein hydrolysis using proteases:An important tool for food biotechnology[J].Journal of Molecular Catalysis B:Enzymatic, 2013, 90:1-11.
[6] ZEEB B, MCCLEMENTS D J, WEISS J.Enzyme-based strategies for structuring foods for improved functionality[J].Annual Review of Food Science and Technology, 2017, 8:21-34.
[7] 江连洲, 佟晓红, 刘宝华, 等.酶种类对生物解离大豆蛋白酶解物功能性和苦味的影响[J].农业机械学报, 2018, 49(8):368-374.
JIANG L Z, TONG X H, LIU B H, et al.Effect of enzymatic species on function and bitterness of enzyme-assisted aqueous extraction soybean protein hydrolysates[J].Transactions of the Chinese Society for Agricultural Machinery, 2018, 49(8):368-374.
[8] 周亭屹, 高新昌, 党亚丽, 等.基于生物信息技术的生物活性肽研究进展[J].食品工业科技, 2019, 40(12):335-340.
ZHOU T Y, GAO X C, DANG Y L, et al.Research development of the bioactive peptides based on bioinformatics[J].Science and Technology of Food Industry, 2019, 40(12):335-340.
[9] GASTEIGER E, HOOGLAND C, GATTIKER A, et al.Protein Identification and Analysis Tools on the ExPASy Server[M].Totowa:Humana Press, 2005.
[10] 于志鹏, 樊玥, 赵文竹, 等.鸡蛋蛋白ACE抑制肽的筛选、鉴定及其作用机制[J].食品科学, 2020, 41(12):129-135.
YU Z P, FAN Y, ZHAO W Z, et al.Identification and mechanism of action of angiotensin-I converting enzyme inhibitory peptides from hen egg proteins[J].Food Science, 2020, 41(12):129-135.
[11] BLEAKLEY S, HAYES M, O’SHEA N, et al.Predicted release and analysis of novel ACE-I, renin, and DPP-IV inhibitory peptides from common oat (Avena sativa) protein hydrolysates using in silico analysis[J].Foods, 2017,6(12):108.
[12] 舒文涛, 耿臻, 李金花, 等.大豆7S球蛋白β-伴大豆球蛋白亚基的氨基酸序列分析[J].中国农学通报, 2018, 34(26):48-51.
SHU W T, GENG Z, LI J H, et al.Amino acid sequence analysis of 7S globulin (β-conglycinin) of soybean[J].Chinese Agricultural Science Bulletin, 2018, 34(26):48-51.
[13] DEAK N A, MURPHY P A, JOHNSON L A.Characterization of fractionated soy proteins produced by a new simplified procedure[J].Journal of the American Oil Chemists′ Society, 2007, 84(2):137-149.
[14] ADLER-NISSEN J.Enzymic Hydrolysis of Food Proteins[M].London:Applied Science Publishers, 1986.
[15] 陈林, 吴克刚, 柴向华, 等.微射流均质预处理提高大豆分离蛋白酶解效率及酶解产物乳化性能[J].农业工程学报, 2015, 31(5):331-338.
CHEN L, WU K G, CHAI X H, et al.Microfluidization pretreatment improving enzymatic hydrolysis of soy isolated protein and emulsifying properties of hydrolysates[J].Transactions of the Chinese Society of Agricultural Engineering, 2015, 31(5):331-338.
[16] 姜振峰, 赫卫, 汪洋, 等.大豆种子7S、11S球蛋白及7S球蛋白亚基的研究[J].中国油料作物学报, 2007, 29(2):138-141.
JIANG Z F, HE W, WANG Y, et al.Study on 7S, 11S globulin and subunits of 7S globulin of soybean seed[J].Chinese Journal of Oil Crop Sciences, 2007, 29(2):138-141.
[17] TSUMURA K, SAITO T, TSUGE K, et al.Functional properties of soy protein hydrolysates obtained by selective proteolysis[J].LWT-Food Science & Technology, 2005, 38(3):255-261.
[18] KILARA A, PANYAM D.Peptides from milk proteins and their properties[J].Critical Reviews in Food Science and Nutrition, 2003, 43(6):607-633.
[19] DICKINSON E.Hydrocolloids as emulsifiers and emulsion stabilizers[J].Food Hydrocolloids, 2009, 23, 1473-1482.
[20] MENGUAL O, MEUNIER G, CAYRÉ I, et al.TURBISCAN MA 2000:Multiple light scattering measurement for concentrated emulsion and suspension instability analysis[J].Talanta, 1999, 50(2):445-456.
[21] NASTAJ M, TERPIŁOWSKI K, SOŁOWIEJ B G.The effect of native and polymerised whey protein isolate addition on surface and microstructural properties of processed cheeses and their meltability determined by Turbiscan[J].International Journal of Food Science and Technology, 2020, 55(5):2179-2187.
[22] 吴彤, 王瑞明, 黄磊, 等.蛋白酶K的研究进展[J].食品工业科技, 2013(14):380-384.
WU T, WANG R M, HUANG L, et al.Research progress in the Proteinase K[J].Science and Technology of Food Industry, 2013(14):380-384.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.