橄榄仁抗氧化肽的分离鉴定及分子对接  

Isolation,identification and molecular docking of antioxidant peptides from olive kernel

作　　者：张迎阳[1] 董亚云 邹平[1] 郑文武 ZHANG Yingyang;DONG Yayun;ZOU Ping;ZHENG Wenwu(School of Pharmacy and School of Biology and Food Engineering,Changzhou University,Changzhou 213164,China)

机构地区：[1]常州大学药学院/生物与食品工程学院,江苏常州213164

出　　处：《食品与发酵工业》2024年第5期189-198,共10页Food and Fermentation Industries

基　　金：江苏省科技项目(BY20212150);常州市科技项目(CE20212030)。

摘　　要：该研究利用酶解法结合凝胶色谱柱从橄榄仁中分离纯化得到6个抗氧化肽,采用分子对接模拟6种抗氧化肽与超氧化物歧化酶(superoxide dismutase, SOD)的结合机制。采用酶解法从橄榄仁中提取抗氧化活性肽,以水解度、DPPH自由基清除率为指标,从6种商用酶中筛选出最适蛋白酶为木瓜蛋白酶,以底物浓度、酶底比、酶解时间、温度进行单因素试验,选取影响酶解反应的3个因素(酶底比>温度>时间)进行响应面法优化得出最佳工艺条件为底物质量浓度50 g/L、酶底比为3.3%(质量分数)、温度20℃、时间88 min。对最优条件下的粗肽液进行SephadexG-25和SephadexG-15分离纯化得到6个具有良好活性的抗氧化肽,分别为WLSF、TSVLR、LVYLVR、AGGKPFQ、YYPFKGFA、AFNVDERLAR。TSVLR显示出最高的DPPH自由基清除活性81.07%和羟自由基清除能力42.48%。分子对接研究表明,纯化的肽通过疏水效应和氢键与抗氧化相关的关键蛋白SOD有效结合。结果表明,橄榄仁可作为天然食物来源制取功能性食品的一个新的探究方向。In this study,six antioxidant peptides were isolated and purified from the olive kernel by enzymatic hydrolysis combined with gel chromatography.Molecular docking was used to simulate the binding mechanism of six antioxidant peptides with the superoxide dismutase(SOD)enzyme.The antioxidant peptides were extracted from the olive kernel by enzymatic hydrolysis.Taking the degree of hy-drolysis and the scavenging rate of DPPH free radicals as indexes,the optimum protease was selected as papain from six commercial en-zymes.The single-factor test was carried out with substrate concentration,enzyme bottom ratio,and enzymatic hydrolysis time and temper-ature.Three factors affecting the enzymatic hydrolysis reaction(enzyme bottom ratio>temperature>time)were optimized by response sur-face methodology,and the optimum process conditions were obtained as follows:substrate concentration of 50 g/L,enzyme bottom ratio of 3.3%,the temperature of 61℃,time of 88 min.Six antioxidant peptides with good activity were isolated and purified by SephadexG-25 and SephadexG-15 from the crude peptide solution under optimal conditions.They were WLSF,TSVLR,LVYLVR,AGGKPFQ,YYPFKGFA,and AFNVDERLAR.TSVLR showed the highest DPPH radical scavenging activity of 81.07%and·OH radical scavenging activity of 42.48%.Molecular docking studies showed that the purified peptide effectively bound to the key protein SOD related to antioxi-dation through hydrophobic effect and hydrogen bond.The results show that olive kernel can be used as a natural food source to produce functional food.

关 键 词：橄榄仁 抗氧化肽 响应面优化 分离鉴定 分子对接 

分 类 号：TS201.2[轻工技术与工程—食品科学]