紫苏粕蛋白抗氧化活性肽的制备、分离纯化及序列鉴定  

Preparation,Isolation,Purification and Sequence Identification of Antioxidant Peptide from Perilla Meal Protein

作　　者：张恒慧[1,3] 张志军 陈士国 叶兴乾[3] 张国华[4] Zhang Henghui;Zhang Zhijun;Chen Shiguo;Ye Xingqian;Zhang Guohua(Biology and Food Laboratory,Taiyuan Institute of Technology,Taiyuan 030008;School of Chemical Engineering and Technology,North University of China,Taiyuan 030051;College of Biosystems Engineering and Food Science,Zhejiang University,Hangzhou 310058;School of Life Science,Shanxi University,Taiyuan 030006)

机构地区：[1]太原工业学院生物与食品教研室,太原030008 [2]中北大学化学工程与技术学院,太原030051 [3]浙江大学生物系统工程与食品科学学院,杭州310058 [4]山西大学生命科学学院,太原030006

出　　处：《中国食品学报》2023年第9期347-355,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：山西省高等学校科技创新项目(2020L0625);山西省应用基础研究项目(201801D221279);教育部中西部高等学校青年骨干教师国内访学项目;山西现代农业产业技术体系建设项目(2021-09)。

摘　　要：以紫苏粕粉为原料,使用碱性蛋白酶进行水解反应,制备抗氧化活性肽。采用超滤离心、凝胶过滤色谱和反相色谱等分离纯化手段对其富集。结果表明,相较其它3种蛋白酶,碱性蛋白酶Alcalase对紫苏粕蛋白的水解程度最高,约为(25.94±0.21)%;碱性蛋白酶作用紫苏蛋白后的酶解产物的抗氧化活性最好,对DPPH自由基清除率约为(91.01±0.73)%。酶解上清液经超滤离心分离后最小分子质量组分F1(小于3 ku)抗氧化活性最强,F1组分经Sephadex G-25凝胶过滤色谱分离后按照分子质量大小依次得到P1、P2、P33个组分,其中分子质量最小的P3组分抗氧化活性最高。P3组分经反相色谱分离所得4个组分中,最后被洗脱出来的P3-4组分疏水性最强且DPPH·自由基清除率最高,质量浓度为3μg/mL的P3-4溶液的DPPH·清除率为(58.8±0.78)%。通过LC-MS-MS鉴定,抗氧化活性肽P3-4组分为十二肽,其氨基酸序列为Lys-Leu-Lys-Asp-Ser-Phe-Glu-Arg-Gln-Gly-Met-Val,分子质量为1437.8 u。本研究结果为深入开发紫苏蛋白资源,研发紫苏抗氧化活性肽提供理论参考。Antioxidant peptides were prepared from Perilla meal protein by hydrolysis with Alcalase,and were enriched by ultrafiltration centrifugation,gel filtration chromatography and reverse phase chromatography.The results showed that compared with the other three proteases,the hydrolysis degree of alcalase to perilla meal protein was the highest,about(25.94±0.21)%.The hydrolysate of perilla meal protein treated by alcalase had the best antioxidant activity,and the scavenging rate of DPPH free radical was about(91.01±0.73)%.The smallest molecular weight component F1(less than 3 ku)of hydrolysate supernatant separated by ultrafiltration and centrifugation had the strongest antioxidant activity.The F1 component was separated by Sephadex G-25 gel filtration chromatography to obtain three components P1,P2 and P3 in order of molecular weight,among which P3 component with the smallest molecular weight had the highest antioxidant activity.Among the four components of P3 separated by reversed-phase chromatography,P3-4 eluting was the most hydrophobic and had the highest DPPH·scavenging capacity,and the DPPH·scavenging rate of P3-4 with concentration of 3μg/mL was(58.8±0.78)%.By LC-MS-MS identification,the antioxidant peptide in component P3-4 was confirmed as a dodeceptide,whose amino acid sequence was Lys-Leu-Lys-Asp-Ser-Phe-Glu-Arg-Gln-GlY-Met-Val,and the molecular weight was 1437.8 u.The results of this study provide a theoretical reference for further development of Perilla protein resources and research of Perilla antioxidant peptides.

关 键 词：紫苏粕蛋白 抗氧化 分离纯化 生物活性肽 氨基酸序列 

分 类 号：TS229[轻工技术与工程—粮食、油脂及植物蛋白工程]