亚麻籽粕制备小分子抗氧化活性肽  

Preparation of Small Molecular Antioxidant Peptides from Flaxseed Meal

作　　者：张文敏 张健 周浩纯 赵迪 李赫 刘新旗 肖林 ZHANG Wenmin;ZHANG Jian;ZHOU Haochun;ZHAO Di;LI He;LIU Xinqi;XIAO Lin(Beijing Advanced Innovation Center for Food Nutrition and Human Health,Beijing Higher Institution Engineering Research Center of Food Additives and Ingredients,School of Food Chemical Engineering,Beijing Technology and Business University,Beijing 100048,China;Shandong Longlive Bio-Technology Co.Ltd.,Yucheng 251200,China)

机构地区：[1]北京食品营养与人类健康高精尖创新中心,北京市食品添加剂工程技术研究中心,北京工商大学食品学院,北京100048 [2]山东龙力生物科技股份有限公司,山东禹城251200

出　　处：《食品科学》2020年第8期36-44,共9页Food Science

基　　金：“十三五”国家重点研发计划重点专项(2016YFD0400401);北京市人才培养质量建设-一流专业建设(市级)-食品科学与工程项目(PXM2019_014213_000010)。

摘　　要：目的:以亚麻籽粕为原料,提取蛋白并制备抗氧化活性肽。方法:利用碱性蛋白酶、中性蛋白酶和胰蛋白酶酶解蛋白,并通过测定总抗氧化能力(Ferric ion reducing antioxidant power,FRAP)、羟自由基清除能力和2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)(2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS)阳离子自由基清除能力表征酶解产物的活性。使用十二烷基硫酸钠-聚丙烯酰氨凝胶电泳和凝胶色谱法分析蛋白和酶解产物的分子质量(mw)分布。利用高效液相色谱-串联质谱联用技术分析鉴定最具有抗氧化活性组分。结果表明,对于高分子质量(3~10 kDa)和低分子质量(<3 kDa)的蛋白多肽,低分子质量多肽(碱性蛋白酶)的抗氧化活性优于其他。10 mg/mL低分子质量多肽(碱性蛋白酶)的FRAP相当于(1.77±0.03)mmol/L的FeSO4,羟自由基清除率为(57.13±1.47)%,清除ABTS阳离子自由基的能力(以Trolox当量浓度计算)为(1.19±0.03)mmol/L。通过凝胶色谱分离的最具抗氧化活性组分F2的蛋白质量占整体组分的20%,0.1 mg/mL组分F2的FRAP相当于0.29 mmol/L的FeSO4,占整体组分活性的45.3%。亚麻籽粕蛋白氨基酸组成齐全,含人体所需的8种必需氨基酸,是一种优质的植物蛋白;蛋白酶解的特异性和肽段分子质量共同影响亚麻籽蛋白多肽的抗氧化活性;亚麻籽粕可以制备高抗氧化活性多肽。Objective:To extract protein from flaxseed meal and then use it to prepare antioxidant active peptides.Methods:Flaxseed protein was digested with alcalase,neutral protease or trypsin,and the antioxidant activities of the hydrolysates were characterized by measuring total antioxidant capacity,Ferric ion reducing antioxidant power(FRAP),hydroxyl radical scavenging ability and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)radical cation scavenging ability.The molecular mass(mw)distribution of the protein and its hydrolysates were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and gel permeation chromatography.The most antioxidant-active components were identified by HPLC-MS/MS analysis.Results:The smaller peptides(<3 kDa)isolated from the alcalase hydrolysate had stronger antioxidant activity than did the larger ones(3–10 kDa).The total antioxidant capacity of the smaller peptides at 10 mg/mL was equivalent to(1.77±0.03)mmol/L FeSO4,the hydroxyl radical scavenging rate was(57.13±1.47)%,and the ability to scavenge ABTS radical cation was equivalent to(1.19±0.03)mmol/L Trolox equivalent concentration.The protein content of fraction F2 with the strongest antioxidant activity separated by gel permeation chromatography accounted for 20%of the total amount.The antioxidant activity of F2 at 0.1 mg/mL was equivalent to 0.29 mmol/L FeSO4,accounting for 45.3%of the total.The flaxseed meal protein contained a full range of amino acids including the eight essential ones and thus could be considered as a high-quality plant protein.The antioxidant activity of its hydrolysates seemed to be affected by the specificity of the hydrolases and the molecular mass of the peptides.Flaxseed meal can be used to produce high antioxidant-active peptides.

关 键 词：亚麻籽粕蛋白 亚麻籽蛋白多肽 抗氧化活性 碱性蛋白酶 

分 类 号：TS201[轻工技术与工程—食品科学]