食品与机械2024，Vol.40Issue(5)：51-61,11.DOI:10.13652/j.spjx.1003.5788.2024.60028
核桃粕源抗氧化活性肽的酶解制备及活性分析
Enzymatic hydrolysis preparation and activity analysis of antioxidant peptides derived from walnut dregs
摘要
Abstract
Objective:This study focused on the identification and activity analysis of antioxidant active peptides in walnut dregs protein hydrolysates.Methods:Protein was extracted from walnut meal by fractionation method,and five kinds of proteolytic enzymes,including trypsin,papain,neutral protease,alkaline protease and flavor protease,were used to hydrolyze the proteins.The antioxidant activity of trypsin hydrolysate(TH),papain hydrolysate(PH),dispase hydrolysate(DH),alcalase hydrolysate(AH)and flavourzyme hydrolysate(FPH)were evaluated and compared.The in vitro antioxidant activity of different concentrations of AH was determined by the detection indexes of DPPH free radical scavenging rate,ABTS free radical scavenging rate and hydroxyl free radical scavenging rate.The sequences of all peptides in AH were identified by LC-MS/MS technology,and the potential antioxidant active peptides of walnut were screened by PeptideRanker,BIOPEP-UWM database and molecular docking tool AutoDock1.5.6,and the two antioxidant active peptides were obtained.The above two peptides were simulated with Keap1 protein,and the antioxidant activity of the two antioxidant peptides was analyzed by using the zebrafish embryo oxidative damage model.Results:The optimal scavenging rates of AH with better antioxidant activity among the five enzymatic hydrolysates were(71.56±0.75)％,(60.63±0.45)％and(98.63±0.06)％,respectively.LC-MS/MS was identified by computer analysis to obtain two active ALWPF and PLRWPF peptides,both of which could bind to the active sites of Keap1 protein,with binding energies of-9.2 and-9.6 kJ/mol,respectively.The safe concentrations of zebrafish embryos treated with ALWPF and PLRWPF peptides ranged from 0～50 μg/mL and 0～30 μg/mL,respectively.Conclusion:Two antioxidant peptides were obtained from walnut meal,and the antioxidant effects of them were verified by zebrafish embryo model.The results of this study can provide a basis for the in-depth utilization of walnut protein.关键词
核桃蛋白/酶解工艺/抗氧化活性/LC-MS/MS/生物信息学分析/斑马鱼胚胎模型Key words
walnut protein/enzymatic hydrolysis process/antioxidant activity/LC-MS/MS/bioinformatics analysis/zebrafish embryo model引用本文复制引用
倪策,曹天红,陈敏,喻勋,吴昊,文李..核桃粕源抗氧化活性肽的酶解制备及活性分析[J].食品与机械,2024,40(5):51-61,11.基金项目
湖南省科技创新计划资助(编号:2023RC3137) （编号:2023RC3137）
国家级大学生创新创业训练计划项目(编号:S202310536041) （编号:S202310536041）
