麦胚清蛋白抗氧化肽的筛选及对细胞氧化损伤的保护作用

doi:10.7506/spkx1002-6630-20200901-012

食品科学 ›› 2021, Vol. 42 ›› Issue (17): 10-18.doi: 10.7506/spkx1002-6630-20200901-012

麦胚清蛋白抗氧化肽的筛选及对细胞氧化损伤的保护作用

张羽，汪芳，翁泽斌，包伊凡，宋海昭，沈新春

（1.南京财经大学食品科学与工程学院，江苏省现代粮食流通与安全协同创新中心，江苏高校粮油质量安全控制及深加工重点实验室，江苏南京 210023；2.南京中医药大学中医学院·中医学院中西医结合学院，江苏南京 210000）

发布日期:2021-09-29
基金资助:
国家自然科学基金青年科学基金项目（31800280）；江苏省自然科学基金项目（BK20181415）；江苏省高等学校自然科学研究项目（18KJB550006）；江苏省研究生科研与实践创新计划项目（KYCX19_1413）

Screening of Wheat Germ Albumin-Derived Antioxidant Peptides and Its Protective Activity against Cellular Oxidative Damage

ZHANG Yu, WANG Fang, WENG Zebin, BAO Yifan, SONG Haizhao, SHEN Xinchun

(1. Collaborative Innovation Center for Modern Grain Circulation and Safety, Key Laboratory of Grains and Oils Quality Control and Processing, College of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing 210023, China; 2. School of Chinese Medicine, School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing 210000, China)

Published:2021-09-29

摘要/Abstract

摘要： 本研究运用超滤与凝胶层析等分离纯化技术逐级分离麦胚清蛋白中性蛋白酶酶解产物，以氧化自由基吸收能力（oxygen radical absorbance capacity，ORAC）与2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonicacid)，ABTS）阳离子自由基清除率为评价指标，筛选具有较高抗氧化活性的产物组分，并以该组分为研究对象，利用液相色谱-串联质谱结合PeptideRanker数据库活性预测技术筛选出具有高抗氧化活性的小肽，并通过高糖诱导血管平滑肌细胞（vascular smooth muscle cells，VSMCs）及H2O2诱导HepG2细胞与VSMCs建立细胞氧化应激模型，评价小肽对细胞氧化损伤的保护作用。结果表明，经超滤与层析筛选出分子质量较小的V组分抗氧化活性最高，液相色谱-串联质谱鉴定出该组分主要由20 个小肽组成，PeptideRanker预测与抗氧化实验验证发现肽序列LNYPPY（765.3 Da）抗氧化活性最高（ORAC 3.01 μmol TE/μmol）、ABTS阳离子自由基清除率95.58%）。肽序列LNYPPY对H2O2诱导的HepG2细胞和VSMCs氧化损伤具有很好的保护作用，但对高糖诱导的VSMCs异常增殖以及活性氧水平上升无明显影响。综上，肽LNYPPY具有良好的体外抗氧化活性，且对H2O2诱导的细胞氧化损伤具有保护作用，研究可为麦胚清蛋白的开发利用提供重要的理论参考。

关键词: 麦胚清蛋白；分离；抗氧化肽；细胞氧化损伤

Abstract: In this study, neutral protease enzymatic hydrolysates of wheat germ albumin were separated step by step using ultrafiltration and chromatography techniques. Oxygen radical absorbance capacity (ORAC) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonicacid (ABTS) cation radical scavenging capacity were used as antioxidant activity evaluation indicators for selecting hydrolysate components with high antioxidant activity. The selected component was served as the initial material to further screen small peptides with high antioxidant activity by liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with PeptideRanker database activity prediction technology. High glucose-induced vascular smooth muscle cells (VSMCs) and H2O2-induced HepG2 and VSMCs were served as oxidative stress injury models to explore the protective effect of obtained small peptides. Results: Component V with small molecular weight obtained by ultrafiltration and chromatography had the highest antioxidant activity. Twenty small peptides were identified with LC/MS-MS analysis from this fraction. PeptideRanker prediction and antioxidant experiment validation showed that the peptide sequence LNYPPY (765.3 Da) had the highest antioxidant activity (ORAC was 3.01 μmol TE/μmol), and ABTS cation radical clearance rate was 95.58%). This peptide had a good protective effect on the H2O2-induced oxidative damage in HepG2 cells and vascular smooth muscle cells (VSMCs), but had no significant effect on the abnormal proliferation and the increase of reactive oxygen species (ROS) levels in VSMCs induced by high glucose. In conclusion, the peptide LNYPPY has good antioxidant properties in vitro and has protective effect against cellular oxidative damage induced by H2O2, which provides an important theoretical reference for the development and utilization of wheat germ albumin.

Key words: wheat germ albumin; isolation; antioxidant peptide; cellular oxidative damage

中图分类号:

TS201.2

张羽，汪芳，翁泽斌，包伊凡，宋海昭，沈新春. 麦胚清蛋白抗氧化肽的筛选及对细胞氧化损伤的保护作用[J]. 食品科学, 2021, 42(17): 10-18.

ZHANG Yu, WANG Fang, WENG Zebin, BAO Yifan, SONG Haizhao, SHEN Xinchun. Screening of Wheat Germ Albumin-Derived Antioxidant Peptides and Its Protective Activity against Cellular Oxidative Damage[J]. FOOD SCIENCE, 2021, 42(17): 10-18.

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麦胚清蛋白抗氧化肽的筛选及对细胞氧化损伤的保护作用

Screening of Wheat Germ Albumin-Derived Antioxidant Peptides and Its Protective Activity against Cellular Oxidative Damage

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