[1]苏子健,孙乐常,颜龙杰,等.黑豆胰蛋白酶抑制剂性质分析及高温对其消化特性的影响[J].大豆科学,2019,38(04):597-606.[doi:10.11861/j.issn.1000-9841.2019.04.0597]
　SU Zi-jian,SUN Le-chang,YAN Long-jie,et al.Purification, Characterization of a Trypsin Inhibitor from Black Soybean ［Glycine max(L.) merr.］ and Effect of High Temperature on Its Digestion[J].Soybean Science,2019,38(04):597-606.[doi:10.11861/j.issn.1000-9841.2019.04.0597]
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黑豆胰蛋白酶抑制剂性质分析及高温对其消化特性的影响

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第38卷 期数: 2019年04期 页码: 597-606 栏目: 出版日期: 2019-07-20
Title:
Purification, Characterization of a Trypsin Inhibitor from Black Soybean ［Glycine max(L.) merr.］ and Effect of High Temperature on Its Digestion
作者:
（1.集美大学 食品与生物工程学院，福建 厦门 361021； 2.水产品深加工技术国家地方联合工程研究中心，福建 厦门 361021)
Author(s):
SU Zi-jian1;  SUN Le-chang1; 2;  YAN Long-jie1;  ZHANG Ling-jing1; 2;  LIU Guang-ming1; 2;  CAO Min-jie1; 2
(1.College of Food and Biological Engineering,Jimei University,Xiamen 361021, China; 2.National & Local Joint Engineering Research Center of Processing Technology for Aquatic Products, Xiamen 361021, China)
关键词:
纯化; 黑豆; 胰蛋白酶抑制剂; 抑制; 消化
Keywords:
Purification;  Black soybean;  Trypsin inhibitor;  Inhibition;  Digestion
DOI:
10.11861/j.issn.1000-9841.2019.04.0597
文献标志码:
A
摘要:
胰蛋白酶抑制剂(trypsin inhibitor, TI)在食品科学和生物技术中具有重要的研究价值。为探讨黑豆胰蛋白酶抑制剂（black soybean trypsin inhibitor, BSTI）的性质及胃肠液消化特性，本研究通过脱脂、热处理、硫酸铵盐析以及阴离子柱层析等方法，从黑豆中纯化得到BSTI。SDS-PAGE显示BSTI呈单一条带，相对分子质量约为21 kDa。在60℃以下与pH2~11的范围内，BSTI呈现较高稳定性。当BSTI与胰蛋白酶的摩尔比达到1时，胰蛋白酶的活力被抑制在15%以下。抑制动力学结果表明，BSTI对胰蛋白酶的抑制属非竞争性抑制，其抑制常数Ki=0.24 nmol?L-1。圆二色谱分析发现，在常温下BSTI的二级结构为：β-折叠43.4%，无规则卷曲29.0%，β-转角21.2%，α-螺旋8.1%。BSTI的变性温度为61±0.9℃。BSTI在模拟胃肠液消化过程中表现出很强的抗酶解作用，但经121℃热处理30 min后，其抗酶解作用显著降低，易被胃蛋白酶、胰蛋白酶和胰凝乳蛋白酶等主要消化酶消化。
Abstract:
Trypsin inhibitor (TI) has significant research value in food science and biotechnology. In order to investigate the properties of black soybean trypsin inhibitor (BSTI) and the digestive properties of gastrointestinal fluids, BSTI was purified from black soybeans by degreasing, heat treatment, ammonium sulfate salting out and anion column chromatography. SDS-PAGE showed a single band of BSTI with a relative molecular mass of approximately 21 kDa. BSTI was stable below 60℃ and in the range of pH2-11. When the molar ratio of BSTI to trypsin reached 1, the activity of trypsin was suppressed to 15% or less. Inhibition kinetics indicated that the inhibition of trypsin by BSTI was non-competitive type, and its inhibition constantKi=0.24 nmol?L-1. The results of circular dichroism analysis showed that the secondary structure of BSTI at room temperature wasβ-sheet 43.4%, random coil 29.0%,β-turn 21.2%,α-helix 8.1% and its denaturation temperature was 61±0.9℃. BSTI was resistant to simulated gastrointestinal fluid digestion. However, after heat treatment at 121℃ for 30 min, the anti-enzymatic action of BSTI was significantly reduced, and was easily hydrolyzed by major digestive enzymes such as pepsin, trypsin and chymotrypsin.

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备注/Memo

收稿日期：2019-02-19

基金项目：国家自然科学基金（31772049）；福建省科技计划（2017N5011）。
第一作者简介：苏子健（1993-），男，硕士，主要从事食品生物技术研究。E-mali:15959452032@163.com。
通讯作者：曹敏杰（1964-），男，博士，教授，主要从事蛋白质化学及水产加工研究。E-mail:mjcao@jmu.edu.cn。

更新日期/Last Update: 2019-07-25