响应面法优化大黄花鱼肉蛋白水解工艺及多肽抗氧化性研究

以大黄花鱼为实验材料,利用酶法水解大黄花鱼肉蛋白制备抗氧化肽。以还原力为响应值,通过单因素结合响应面法对中性蛋白酶酶解大黄花鱼肉蛋白的酶用量、酶解温度、底物浓度以及酶解时间进行了优化,结果表明:四种酶中,中性蛋白酶酶解的酶解液水解度(DH)和还原能力最高。最优酶解工艺条件为酶用量为0.4%、酶解温度45 ℃、底物浓度25.0%、酶解时间7 h、体系pH7.0时,还原力为0.951。酶解液DH为37.51%,超氧阴离子自由基清除力(O₂-·)为82.42%。SDS-PAGE(聚丙烯酰氨凝胶电泳)结果显示,酶解7 h大黄花鱼肉蛋白肌动蛋白完全消失,水解形成肌球蛋白轻链分子量为27、15和6 kDa。

Optimization of the Protein Hydrolysis Process by Response Surface Method and the Antioxidant Properties of Polypeptides in Large Yellow Croaker

The antioxidant peptide was prepared by enzymatic hydrolysis of the protein from the large yellow croaker. Taking reduction force as the response value, the enzymatic dosage, enzymatic hydrolysis temperature, substrate concentration and enzymatic hydrolysis time of neutral protease were optimized by combining single factor and response surface method. The results showed that, among the four enzymes, neutral protease enzymatic hydrolysis had the highest hydrolytic degree (DH) and reducing capacity.The optimal enzymatic hydrolysis process conditions were as follows:The enzyme dosage was 0.4%, the enzymatic hydrolysis temperature was 45℃, the substrate concentration was 25.0%, the enzymatic hydrolysis time was 7 h, and the system pH was 7.0. The reduction force was 0.951.DH of the enzymatic hydrolysate was 37.51%, and the scavenging power (O₂-·) of superoxide anion radical was 82.42%.SDS-PAGE (polyacrylamide gel electrophoresis) showed that actin disappeared completely after 7 h of enzymatic hydrolysis, and the molecular weights of light chain of hydrolyzed myosin were 27, 15 and 6 kDa.