酶解大豆乳清蛋白制备抗氧化肽及其体外抗氧化活性评价  

Antioxidant Peptides Prepared by Enzymatic Hydrolysis of Whey Soy Proteins and Their Antioxidative Activities in Vitro

作　　者：林彦君 刘杨静 曲肖凤 杜媛媛 房文硕 尹艺童 李瑞 LIN Yanjun;LIU Yangjing;QU Xiaofeng;DU Yuanyuan;FANG Wenshuo;YIN Yitong;LI Rui(School of Biological Science,Jining Medical University,Rizhao 276826,China)

机构地区：[1]济宁医学院生物科学学院,山东日照276826

出　　处：《食品工业科技》2023年第20期230-238,共9页Science and Technology of Food Industry

基　　金：日照市自然科学基金(RZ2021ZR21);国家级大学生创新训练计划项目(201910443009);济宁医学院大学生创新训练计划项目(cx2021086)。

摘　　要：为了提高大豆乳清废水中大豆乳清蛋白的应用价值,以大豆乳清蛋白为原料采用酶解法在自制的超声-恒温反应体系中制备抗氧化肽。以总还原力、DPPH自由基清除率和羟基自由基清除率为指标,结合单因素实验和响应面法优化大豆乳清蛋白的酶解工艺。随后,利用超滤法(截留分子量30 kDa)回收水解液中的抗氧化肽,并以抗坏血酸为参照,评价其体外抗氧化活性。结果表明:中性蛋白酶和胃蛋白酶组合最适于大豆乳清蛋白的水解;在此基础上,确定的最佳工艺条件为酶-底物比5000 U/g大豆乳清蛋白、酶解时间6 h和大豆乳清蛋白浓度9.0 mg/mL,所得大豆乳清蛋白水解液的DPPH自由基清除率为62.3%±1.1%。从该水解液中超滤所得抗氧化肽的总还原力、DPPH自由基清除率和羟基自由基清除率的IC_(50)值分别为0.128、0.221和0.185 mg/mL,分别是抗坏血酸IC50值的2.56、40.76、1.47倍,表明抗氧化肽的活性略低于抗坏血酸。因此,大豆乳清蛋白可作为制备高活性抗氧化肽的来源。To improve the application value of whey soy proteins(WSP)in soy whey wastewater,they were used to prepare antioxidant peptides by enzymatic hydrolysis in a self-made ultrasound-thermostatic reaction system.Using sing-factor tests and response surface methodology,the technology for enzymatic hydrolysis of WSP was optimized with total reducing force,scavenging rate of DPPH radical and scavenging rate of hydroxyl radical as indicators.Then,ultrafiltration with a molecular weight cut-off of 30 kDa was adopted to separate antioxidant peptides from the hydrolysates and ascorbic acid as a reference to evaluate their antioxidant activities in vitro.The results showed that neutral protease and pepsin were the most suitable for the hydrolysis of WSP.On this basis,the optimal technological conditions were determined as enzyme-substrate ratio 5000 U/g WSP,hydrolysis time 6 h and WSP concentration 9.0 mg/mL.Under these conditions,the scavenging rate of DPPH of WSP hydrolysates reached 62.3%±1.1%.The antioxidant peptides isolated from the above hydrolysates had IC50 values for total reducing force,scavenging rate of DPPH radical and scavenging rate of hydroxyl radical of 0.128,0.221 and 0.185 mg/mL,respectively.These corresponding IC_(50) values were 2.56,40.76 and 1.47 times of those of ascorbic acid,respectively.It was indicated that the antioxidant peptides had slightly lower activity than ascorbic acid.Therefore,WSP is a suitable source of antioxidant peptides with high activities.

关 键 词：大豆乳清蛋白 抗氧化肽 酶解 工艺优化 

分 类 号：TS201.2[轻工技术与工程—食品科学]