三酶法制备罗非鱼鱼皮胶原蛋白抗氧化肽及活性研究

Three-Step Enzymatic Hydrolysis of Tilapia Skin Collagen for Preparation of Antioxidant Peptides and Their Antioxidant Activity

为了探索制备罗非鱼鱼皮胶原蛋白抗氧化肽的最佳工艺,在碱性蛋白酶和胰蛋白酶酶解的基础上,以酶解温度、酶解时间、酶解pH值和酶与底物质量比([E]/[S])为试验因素,以水解度和DPPH自由基清除率为响应值,采用响应面分析法优化Orientase 20A酶的酶解条件。结果表明:最优工艺参数为:酶解温度为41.74℃、pH3.97、酶解时间为6h、[E]/[S]为1.5%,酶解液水解度和DPPH自由基清除率的预测值分别为9.42%和35.01%,验证值分别为9.57%和35.21%。响应面对酶解法制备罗非鱼鱼皮胶原蛋白抗氧化肽的酶解条件的优化是可行的,可以用于实际预测。抗氧化活性实验表明,酶解肽具有较强的清除DPPH自由基和ABTS+.能力,其IC50值分别为10.78mg/mL和8.26mg/mL。

This study explored optimal conditions for the enzymatic preparation of antioxidant peptides from tilapia skin collagen and the antioxidant activity of antioxidant peptides.Sequential hydrolysis with alkaline protease,trypsin and Orientase 20A was the best way to prepare antioxidant peptides from tilapia skin collagen,as shown by a comparison of the effects of single use and sequential combinations of proteases on DPPH radical scavenging activity of hydrolyzed tilapia skin collagen.In the final step（Orientase 20A） of the hydrolysis process,process parameters including temperature,time,pH and enzyme/substrate ratio were optimized using response surface methodology based on degree of hydrolysis（DH） and DPPH radical scavenging activity of hydrolyzed tilapia skin collagen to be 41.74 ℃,3.97,6 h and 1.5%,respectively.Under these hydrolysis conditions,the predicted DH and DPPH radical scavenging activity of hydrolyzed tilapia skin collagen were 9.42% and 35.01%,respectively,and the observed values were 9.57% and 35.21%,respectively.Thus,the developed response surface model is feasible and can be used for practical prediction.The tilapia skin collagen hydrolysate obtained under the optimized hydrolysis conditions had potent scavenging activity against DPPH and ABTS＋radicals with an IC50of 10.78 mg/mL and 8.26 mg/mL,respectively.