虾壳源蛋白水解物降糖降脂活性评价及肽序分析  

Study on hypoglycemic and lipid-lowering activity of shrimp shell-derived enzymatic hydrolysate and peptide sequence function analysis

作　　者：韩鹏薇 易彤 李虹辉 吴昊[1] 李彦[1] 褚武英[2] HAN Pengwei;YI Tong;LI Honghui;WU Hao;LI Yan;CHU Wuying(School of Food Science and Bioengineering,Changsha University of Science&Technology,Changsha,Hunan 410114,China;College of Biology and Chemical Engineering,Changsha University,Changsha,Hunan 410022,China)

机构地区：[1]长沙理工大学食品与生物工程学院,湖南长沙410114 [2]长沙学院生物与化学工程学院,湖南长沙410022

出　　处：《食品与机械》2024年第4期148-157,共10页Food and Machinery

基　　金：湖南省教育厅科学研究优秀青年项目(编号:22B0327);湖南省教育厅重点科学研究项目(编号:22A0237);国家自然科学基金青年项目(编号:32102816);湖南省科技创新计划资助(编号:2023RC3137)。

摘　　要：目的:采用酶解法制备克氏原螯虾壳蛋白水解物(Procambarus clarkii shell protein hydrolysates,PCSPHs),并分析其体外降糖降脂活性及肽序。方法:分别采用胃蛋白酶、碱性蛋白酶、胰蛋白酶、风味蛋白酶和木瓜蛋白酶水解制备不同虾壳蛋白水解物,分析其体外降糖降脂活性和肽序列;运用Peptide Ranker及BIOPEP-UWM网站在线分析,再以核受体PPARγ配体结合区域的晶体结构作为靶点,使用Autodock vina进行分子对接模拟,获得具有潜在降糖降脂活性的虾壳肽。结果:胃蛋白酶水解物(PEP-PCSPHs)对α-淀粉酶和α-葡萄糖苷酶活性具有较强的抑制作用,IC 50值分别为(5.42±0.05),(7.11±1.01)mg/mL;胰蛋白酶水解物(TRY-PCSPHs)对胰脂肪酶活性具有最强的抑制能力,IC 50值为(4.71±1.12)mg/mL,且对甘氨胆酸钠表现出最好的体外结合效果。此外,经质谱鉴定PEP-PCSPHs和TRY-PCSPHs中分别得到3391,2086条肽序;通过在线网站预测和分子对接筛选出多条均能与PPARγ结合的降糖降脂虾壳活性肽(PCSAPs)。结论:酶解克氏原螯虾壳制备的虾壳蛋白水解物具有潜在的降糖降脂活性,可能改善糖脂代谢紊乱。Objective:The PCSPHs were prepared by enzymatic hydrolysis of Procambarus clarkii shells,and their hypoglycemic and lipid-lowering activities in vitro were evaluated and peptide sequence were analyzed.Methods:Different crayfish shell proteolysates were prepared by hydrolysis of pepsin,alcalase protease,trypsin,flavor protease and papain,and their in vitro hypoglycemic and lipid-lowering activities were evaluated and peptide sequences were determined.The peptides sequence of Procambarus clarkii shells was identified by LC-MS/MS.Taking the crystal structure of the nuclear receptor PPARγligand binding region as the target,Autodock vina was used to simulate molecular docking to obtain crayfish shell peptides with potential hypoglycemic/lipid-lowering activities.Results:The PEP-PCSPHs had significant inhibitory effects onα-amylase andα-glucosidase activity,with IC 50 values of(5.42±0.05)mg/mL and(7.11±1.01)mg/mL,respectively.The TRY-PCSPHs had the strongest inhibitory effect on pancreatic lipase activity,with an IC 50 of(4.71±1.12)mg/mL,and exhibited the best in vitro binding effects on sodium glycinocholate.In addition,3391 peptide sequences were identified in pepsin hydrolysates and 2086 peptide sequences were identified in trypsin hydrolysates,and multiple hypoglycemic/lipid-lowering crayfish shell active peptides that could bind to PPARγwere screened through online website prediction and molecular docking.Conclusion:The shrimp shell peptides prepared by enzymatic hydrolysis of crayfish shells have potential hypoglycemic and lipid-lowering activities,which may play a role in improving glucose and lipid metabolism disorders.

关 键 词：克氏原螯虾壳 蛋白水解物 PPARΓ 分子对接 降糖降脂活性 

分 类 号：TS254.9[轻工技术与工程—水产品加工及贮藏工程]