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复合蛋白酶水解黑豆粕制备多肽的工艺优化
Optimization of hydrolysis process of black soybean meal by composite proteases for preparation of polypeptides
  
DOI：10.12166/j.zgyz.1003-7969/2019.11.015
中文关键词:  为充分利用黑豆粕中的蛋白质资源，研究了复合蛋白酶水解黑豆粕制备多肽的工艺路线。从蛋白酶的筛选入手，考察了不同蛋白酶对黑豆粕水解效果的差异，并通过复配比例优化了最佳复合蛋白酶的组成。采用单因素试验及响应面试验确定了复合蛋白酶水解黑豆粕的最佳工艺条件。结果表明：最佳复合蛋白酶为胰蛋白酶、碱性蛋白酶与桔青霉蛋白酶按酶活比1∶ 3∶ 2复配  最佳工艺条件为加酶量5 670 U/g，底物蛋白质量浓度40 g/L，pH 9.55，水解温度50 ℃，水解时间6 h。在优化的工艺条件下水解黑豆粕，多肽得率可以达到82.44%。水解液中多肽的相对分子质量均在7.8 kDa以下，大部分多肽的相对分子质量小于3.4 kDa  多肽基本上保持了黑豆粕蛋白的氨基酸组成，具有较高的营养价值。
英文关键词:：black soybean meal  composite protease  hydrolysis  polypeptide
基金项目:广东省公益研究与能力建设专项资金项目（2015 A010107016，2016A010105024）；国家星火计划项目（2013GA 780084）
Author NameAffiliation
PAN Jinquan, JIANG Bian, ZHANG Enshu, LI Huayan School of Life Science and Technology, Lingnan Normal University, Zhanjiang 524048, Guangdong, China 
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中文摘要:
      为充分利用黑豆粕中的蛋白质资源，研究了复合蛋白酶水解黑豆粕制备多肽的工艺路线。从蛋白酶的筛选入手，考察了不同蛋白酶对黑豆粕水解效果的差异，并通过复配比例优化了最佳复合蛋白酶的组成。采用单因素试验及响应面试验确定了复合蛋白酶水解黑豆粕的最佳工艺条件。结果表明：最佳复合蛋白酶为胰蛋白酶、碱性蛋白酶与桔青霉蛋白酶按酶活比1∶ 3∶ 2复配；最佳工艺条件为加酶量5 670 U/g，底物蛋白质量浓度40 g/L，pH 9.55，水解温度50 ℃，水解时间6 h。在优化的工艺条件下水解黑豆粕，多肽得率可以达到82.44%。水解液中多肽的相对分子质量均在7.8 kDa以下，大部分多肽的相对分子质量小于3.3 kDa；多肽基本上保持了黑豆粕蛋白的氨基酸组成，具有较高的营养价值。
英文摘要:
      In order to make full use of the protein in black soybean meal, the hydrolysis process of black soybean meal using composite proteases for preparation of polypeptides was investigated. The hydrolysis effects of different proteases on black soybean meal were compared, and the optimal composite protease composition was determined by optimizing the mixing proportion. The hydrolysis process conditions were optimized by single factor experiment and response surface methodology. The results showed that the optimal composite protease was composed of trypsin, alkaline protease and the protease from Penicillium citrinum in a proportion of 1∶ 3∶ 2. The optimal process conditions were obtained as follows: enzyme dosage 5 670 U/g, substrate protein mass concentration 40 g/L, pH 9.55, enzymatic hydrolysis temperature 50 ℃ and enzymatic hydrolysis time 6 h. Under these conditions, the yield of polypeptides could reach 82.44%. The relative molecular weights of polypeptides in hydrolysate were below 7.8 kDa, and most of them were less than 3.3 kDa. Polypeptides basically maintained the amino acid composition of protein from black soybean meal, and had high nutritional value.
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