大黄鱼鱼肉蛋白水解制备抗氧化肽的工艺

Enzymatic preparation of antioxidant peptides from large yellow croaker

以大黄鱼为原料,采用不同蛋白酶(碱性蛋白酶、中性蛋白酶、木瓜蛋白酶、胃蛋白酶、胰蛋白酶和复合蛋白酶)分别水解鱼肉,制备抗氧化肽;研究不同酶的水解效果,筛选出最佳的水解用酶;采用单因素试验,以蛋白水解度及水解液的还原力和超氧阴离子自由基(O-·2)清除力为指标,研究不同水解p H、酶添加量、水解温度、底物浓度和水解时间对水解效果的影响;在此基础上,采用Box-Behnken响应面设计法,以水解液的还原力为评价指标,对水解工艺参数进行优化.结果表明,中性蛋白酶为水解大黄鱼的最适用酶,其最佳工艺条件为:底物浓度26.55%、水解温度46℃、水解时间7.2 h、p H 7.0、酶添加量2400 U·g-1,在此条件下,水解液的还原力达到最大(0.967),此时,O-·2清除力为87.46%,蛋白水解度为36.51%.

In order to obtain the antioxidant peptides,proteases were used to hydrolyze large yellow croaker( Pseudosciaena crocea)and the hydrolytic process was optimized. We compared the hydrolysis efficiencies of Alcalase,Neutrase,Papain,Pepsin,Trypsin and Protamex. By measuring the hydrolysis degree and reducing ability,the hydrolytic enzyme was selected. Then the hydrolysis p H,hydrolysis temperature,enzyme dose,substrate concentration and hydrolysis time were studied in single factor test. On the basis,Box-Behnken experimental design combined with response surface methodology was employed to explore the optimum conditions. The results showed that Neutrase was the optimal hydrolytic enzyme and the best conditons were determined as follows: substrate concentration 26.55%,hydrolysis temperature 46 ℃,hydrolysis time 7.2 h,p H 7.0 and enzyme dose 2400 U·g-1. These conditions gave a hydrolysis degree of large yellow croaker of 36.51%,and the reducing ability and superoxide anion free radical scavenging ability were 0.967 and 87.46%,respectively.