响应面法优化秋刀鱼酶解制备抗氧化活性肽的工艺  

The Optimization of the Preparation Process of Antioxidative Active Peptide from the Enzymatic Hydrolysate of Pacific Saury(Cololabis saira)by Response Surface Methodology

作　　者：栾俊家 张尚悦[2] 李昂达 李学鹏 励建荣 林洪[3] 王明丽 郭晓华 于建洋 周小敏 LUAN Junjia;ZHANG Shangyue;LI Angda;LI Xuepeng;LI Jianrong;LIN Hong;WANG Mingli;GUO Xiaohua;YU Jianyang;ZHOU Xiaomin(College of Food Science and Engineering,Bohai University,National&Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products,National R&D Branch Center of Surimi and Surimi Products Processing,Jinzhou 121013,China;Department of Navigation,PLA Dalian Naval Academy,Dalian 116018,China;College of Food Science and Technology,Ocean University of China,Qingdao 266100,China;Penglai Jinglu Fishery Co.,Ltd.,Yantai 265600,China;Shandong Meijia Group Co.,Ltd.,Rizhao 276815,China;Rongcheng Taixiang Food Products Co.,Ltd.,Weihai 264300,China;Zhejiang Xingye Industrial Group Co.,Ltd.,Zhoushan 316120,China)

机构地区：[1]渤海大学食品科学与工程学院,生鲜农产品贮藏加工及安全控制技术国家地方联合工程研究中心,国家鱼糜及鱼糜制品加工技术研发分中心,辽宁锦州121013 [2]海军大连舰艇学院航海系,辽宁大连116018 [3]中国海洋大学食品科学与工程学院,山东青岛266100 [4]蓬莱京鲁渔业有限公司,山东烟台265600 [5]山东美佳集团有限公司,山东日照276815 [6]荣成泰祥食品股份有限公司,山东威海264300 [7]浙江兴业集团有限公司,浙江舟山316120

出　　处：《食品工业科技》2022年第5期172-181,共10页Science and Technology of Food Industry

基　　金：国家重点研发计划项目(2016YFD0400705);辽宁省兴辽英才计划项目(XLYC1807133)。

摘　　要：采用酶解法从秋刀鱼肌肉中提取抗氧化活性肽,以水解度(DH)、TCA-可溶性肽含量、DPPH自由基清除率、Fe^(3+)还原力、·OH清除率以及O_(2)^(-)·清除率为指标,从六种商业用酶中(中性蛋白酶、碱性蛋白酶、风味蛋白酶、木瓜蛋白酶、菠萝蛋白酶、胰蛋白酶)筛选出最适蛋白酶。以料液比、酶添加量、酶解时间、温度、pH五个因素进行单因素实验,选取影响酶解显著的三个因素进行响应面优化。并对最优条件下的酶解液进行分子量分布的测定和游离氨基酸含量的测定。结果显示:最适蛋白酶为中性蛋白酶。三因素对酶解液抗氧化活性的影响大小为料液比>酶添加量>酶解时间,秋刀鱼抗氧化活性肽的最佳制备工艺为料液比为1:2.67(质量体积比,g/mL)、酶添加量为0.60%、酶解时间为4.04 h、温度为50℃、pH7.0,在此条件下制得的酶解液DPPH自由基清除率为92.33%,Fe^(3+)还原力为0.711(20 mg/mL),与预测值无显著性差异(P>0.05)。可见,采用响应面法优化酶解工艺制备的秋刀鱼抗氧化活性肽具有较高的抗氧化活性。最佳酶解工艺条件下制得的酶解液中,分子量<3000 Da的组分占97.87%。共检测出16种游离氨基酸,其中必需氨基酸占总氨基酸含量的39.79%,抗氧化活性氨基酸占总氨基酸含量的37.01%。本文为秋刀鱼的高值化利用和多肽类功能性食品的开发提供了一定的参考。The antioxidative peptides were prepared from the muscle of Pacific saury by enzymatic hydrolysis.Based on the degree of hydrolysis(DH),the content of TCA-soluble peptide,DPPH free radical scavenging rate,Fe^(3+) reducing power,·OH radical scavenging rate and O_(2)^(-)·radical scavenging rate,the optimal protease was selected from six kinds of commercial enzyme(neutral protease,alkaline protease,flavour protease,papain,bromelain and trypsin).Single factor experiments were carried out with five factors:Solid-liquid ratio,the amount of enzyme,enzymatic hydrolysis time,temperature and pH,and three factors(solid-liquid ratio,the amount of enzyme and enzymatic hydrolysis time)were selected to optimize the response surface methodology(RSM).And the molecular weight distribution and free amino acid content of the hydrolysate were determined under the optimal conditions.The results showed that the optimal protease was neutral protease.The influence of three factors on the antioxidant activity of the enzymatic hydrolysate were the solid-liquid ratio>the amount of enzyme>enzymatic hydrolysis time.The optimal preparation processes of the antioxidative active peptide were determined as follows:the solid-liquid ratio was 1:2.67(mass-volume ratio,g/mL),the amount of enzyme was 0.60%,enzymatic hydrolysis time was 4.04 h,temperature was 50℃,and pH7.0.Under these conditions,DPPH free radical scavenging rate and Fe^(3+) reducing power were 92.33%and 0.711(20 mg/mL),respectively,which were no significant difference with the predicted value(P>0.05).It could be seen that the antioxidative peptide peptide optimized by response surface methodology had high antioxidant activity.Under the optimal conditions,97.87%of the components with molecular weight less than 3000 Da were the main components of the hydrolysate.A total of 16 kinds of free amino acids were detected,among which essential amino acids accounted for 39.79%and antioxidant active amino acids accounted for37.01%of the total amino acids.This study provided some reference for

关 键 词：秋刀鱼 酶解 抗氧化活性肽 氨基酸组成 分子量 

分 类 号：TS254.4[轻工技术与工程—水产品加工及贮藏工程]