榛子粕抗氧化肽的分离纯化及序列分析

陈艳; 吕春茂; 韩金晶; 姬昱佳; 付沁璇; 刘璐; 徐嘉一

doi:10.13386/j.issn1002-0306.2018.15.021

榛子粕抗氧化肽的分离纯化及序列分析

Separation,Purification and Sequence Analysis of Antioxidative Peptides from the Halnut Meal Protein Hydrolysate

摘要

摘要: 为研究榛子粕抗氧化肽的抗氧化性和氨基酸组成的关系,本实验通过中性蛋白酶水解榛子粕蛋白得到榛子粕抗氧化肽,以DPPH自由基清除率为指标,采用超滤分离和葡聚糖凝胶纯化,获得DPPH自由基清除率高的榛子粕抗氧化肽P2,采用超高压液相色谱串联四级杆质谱(LC-MS/MS)测定榛子粕抗氧化肽的氨基酸序列。P2中7个短肽的氨基酸序列分别为His-ILe/Leu-Pro(362 Da)、ILe/Leu-Val-Asp-Glu(475 Da)、Thr-Pro-Pro-His-Lys(588 Da)、His-ILe/Leu-His-Ser-Ala-Thr(662 Da)、Cys-His-Ser-ILe/Leu-Met-ILe/Leu(701 Da)、Thr-His-Ala(327 Da)、Phe-ILe/Leu(279 Da)。由此得出结论,疏水性氨基酸和芳香性氨基酸可以提高榛子粕抗氧化肽的抗氧化性。

Abstract: In order to study the relationship between antioxidant activity and amino acid composition of halnut meal antioxidant peptides. The hydrolysate from halnut meal protein was prepared by neutral protease. Halnut meal antioxidant peptides were separated by ultraflitration and gel filtration chromatography(Sephadex G-10),the fractions were evaluated for their antioxidant activity by DPPH free redical scavenging activity.The frations P2 with high antioxidant activity were examined by liquid charomatography-mass spectrometry(LC-MS/MS). Seven novel peptides with weight of 362,475,588,662,701,327,279 Da were identified as His-ILe/Leu-Pro,ILe/Leu-Val-Asp,Thr-Pro-Pro-His-Lys,His-ILe/Leu-His-Ser-Ala-Thr,Cys-His-Ser-ILe/Leu-Met-ILe/Leu,Thr-His-Ala and Phe-ILe/Leu.The conclusion was that hydrophobic amino acids and aromatic amino acids could improve antioxidant activity of halnut meal antioxidant peptides.