基于分子模拟技术筛选牛乳清蛋白源抗氧化肽  

Screening Antioxidant Peptides from Bovine Whey Protein Based on Molecular Simulation Technology

作　　者：马寅龙 郭锐斌 于雁松 马隆凯 许晓曦[1] Ma Yinlong;Guo Ruibin;Yu Yansong;Ma Longkai;Xu Xiaoxi(Key Laboratory of Dairy Science and Education,College of Food Science,North East Agricultural University,Harbin 150030)

机构地区：[1]东北农业大学食品学院、乳品科学教育部重点实验室,哈尔滨150030

出　　处：《中国食品学报》2024年第2期11-31,共21页Journal of Chinese Institute Of Food Science and Technology

基　　金：黑龙江省“百千万”工程科技重大专项(2020ZX07B01-2-5)。

摘　　要：传统抗氧化肽的筛选往往需要经过酶解、纯化、分离、鉴定及体外试验验证,时间和物力花费较大。本研究以生物信息学为理论基础,利用计算机虚拟消化、相关活性预测与评价、药物代谢动力学及分子模拟技术对牛乳清蛋白源抗氧化肽进行依次筛选,对其可能发生的Keap1-Nrf2-ARE抗氧化途径进行分析、解释,分析其分子反应过程中构象的稳定性。应用PeptideRanker程序对生物活性进行预测评分,筛选得到137条评分大于0.4的肽段,同时采用ToxinPred、Innovagen、Expasy-compute及Pepdraw程序进一步分析肽段的理化性质,并通过药物代谢动力学ADMET、TOPKAT分析对肽段进行筛选,得到14条无毒、无致敏性、无致突变性、无致癌性及水溶性良好的多肽,然后将其与Keap1受体进行分子对接。将分子对接评分前4位的多肽与Keap1的最优结合复合物进行构象作用机制分析,最终通过复合物构象分子间作用力评价CDEF序列最具抗氧化活性的潜力,并将CDEF-Keap1受体复合物进行分子动力学模拟,结果显示动力学模拟过程中构象变化较稳定,因此CDEF抗氧化肽可在人体中较好地发挥抗氧化活性。与传统酶解方法相比,本方法可快速、高效筛选抗氧化肽,为天然食品源抗氧化肽的快速筛选提供了新思路。The screening of traditional antioxidant peptides often needs enzymatic hydrolysis,purification,separation,identification and in vitro experimental verification,which costs a lot of time and material resources.Based on bioinformatics,this study used computer virtual digestion,prediction and evaluation of related activities,pharmacokinetics and molecular simulation techniques to screen antioxidant peptides from bovine whey protein in turn,analyze and explain the possible mechanism of Keap1-Nrf2-ARE antioxidant pathway,and analyze the stability of conformation in the process of molecular reaction.Using the Peptideranker program to predict and score the biological activity,137 peptides with a score greater than 0.4 were screened.At the same time,Toxinpred,Innovagen,Expasy-compute and Pepdraw programs were used to further predict and analyze the physical and chemical properties of the peptides.Through pharmacokinetic ADMET and TOPKAT analysis,14 peptides with no toxicity,no sensitization,no mutagenicity,no carcinogenicity and good water solubility were screened.Then it was docked with Keap1 receptor,and the conformational mechanism of the polypeptide with the top four docking scores was analyzed with the optimal binding complex of Keap1.Finally,through the evaluation of the intermolecular force of the complex conformation,it was concluded that the CDEF sequence had the most antioxidant activity potential,and the CDEF-keap1 receptor complex was simulated by molecular dynamics.The results showed that the conformational change was relatively stable in the process of dynamic simulation.Therefore,CDEF antioxidant peptides could play a better role in antioxidant activity in human body.Compared with traditional enzymatic hydrolysis methods,this study could quickly and efficiently screen antioxidant peptides,and provide a new idea for the rapid screening of antioxidant peptides from natural food sources.

关 键 词：乳清蛋白 抗氧化肽 生物活性肽 分子对接 

分 类 号：TS201.2[轻工技术与工程—食品科学]