乳清蛋白水解产物中抗氧化肽的分离纯化及应用  

Isolation,Purification and Applications of Antioxidant Peptides from Whey Protein Hydrolysis

作　　者：王昊乾 刘怡婷 张静雯 张和平[1] 孙天松[1] 姚国强[1] Wang Haoqian;Liu Yiting;Zhang Jingwen;Zhang Heping;Sun Tiansong;Yao Guoqiang(Inner Mongolia Agricultural University,Key Laboratory of Dairy Biotechnology and Engineering,Ministry of Education,Key Laboratory of Dairy Products Processing,Ministry of Agriculture and Rural Affairs,Key Laboratory of Dairy Biotechnology and Engineering,Inner Mongolia Autonomous Region,Hohhot 010018)

机构地区：[1]内蒙古农业大学,乳品生物技术与工程教育部重点实验室,农业农村部奶制品加工重点实验室,内蒙古自治区乳品生物技术与工程重点实验室,呼和浩特010018

出　　处：《中国食品学报》2023年第4期35-47,共13页Journal of Chinese Institute Of Food Science and Technology

基　　金：内蒙古自治区高等学校科学技术研究项目(NJZZ22488);内蒙古自治区自然科学基金项目(2022QN03021);财政部和农业农村部:国家现代农业产业技术体系资助项目(CARS36)。

摘　　要：目的:从乳清蛋白水解物中筛选出一种具有较强抗氧化活性的多肽,减少真空冷冻干燥过程中对乳双歧杆菌Probio-M8的细胞损伤。方法:通过碱性蛋白酶水解乳清蛋白,经超滤分离后得到3种分子质量k1(>8 ku)、k2(3~8 ku)和k3(<3 ku)的多肽样品,以强阳离子交换色谱、反向高效液相色谱(RP-HPLC)、液相色谱质谱联用进行分离、纯化和鉴定,以1,1-二苯基-2-三硝基苯肼(DPPH)清除率、抗超氧阴离子自由基和对氧化自由基的吸收能力作为检测指标,筛选具有较强抗氧化能力的组分作为保护剂,应用于乳双歧杆菌Probio-M8的真空冷冻干燥过程中。结果:经超滤、蛋白层析及液相色谱分离后,k3样品中的3-2和4-6两个组分具有较强抗氧化活性,从组分3-2和4-6中分别鉴定出5条和6条潜在的抗氧化肽序列。在乳双歧杆菌Probio-M8真空冷冻干燥中的应用结果表明,多肽组分3-2能够显著提高菌种存活率至(88.31±0.02)%,同时还能够降低胞内活性氧水平并保护细胞活力。结论:抗氧化多肽组分3-2能够作为冻干保护剂,在真空冷冻干燥过程中减少乳双歧杆菌Probio-M8受到细胞损伤。Objective:A peptide with antioxidant activity was screened from whey protein hydrolysate and used to protect Probio-M8 during the freeze-drying process.The whey proteins were hydrolyzed by alkaline protease and separated by ultrafiltration and three different molecular weight peptide samples were obtained,k1(>8 ku),k2(3-8 ku),and k3(<3 ku).The peptides were then purified and identified by strong cation exchange chromatography,reversed-phase high-performance liquid chromatography(RP-HPLC),and liquid chromatography-mass spectrometry(LC-MS).1,1-Diphenyl-2-trinitrophenylhydrazine(DPPH)scavenging,resistance to superoxide anion radicals,and absorption of oxidative radicals were used as assays to assess antioxidant activity and were screened.Results:Samples 3-2 and 4-6 showed high antioxidant activity.Five and six antioxidant peptide sequences were identified for samples 3-2 and 4-6,respectively.Sample 3-2 was able to protect B.lactis Probio-M8 in vacuum freeze-drying,increasing the survival rate of the strain to(88.31±0.02)%and also reducing intracellular reactive oxygen species levels and protecting cellular enzyme activity.Conclusion:Sample 3-2 was effective in reducing cell damage caused by the vacuum freeze-drying process of B.lactis Probio-M8.

关 键 词：乳清蛋白 抗氧化肽 分离纯化 乳双歧杆菌 真空冷冻干燥 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程]