响应面法优化酶解鸢乌贼制备抗氧化肽的工艺研究

Optimization of enzymatic preparation of antioxidant peptides from purpleback flying squid (sthenoeuthis oualaniensis) protein by response surface methodology

利用响应面分析法对酶法水解鸢乌贼蛋白制备抗氧化肽的工艺条件进行优化。首先通过单因素实验最终确定木瓜蛋白酶为水解鸢乌贼蛋白制备抗氧化肽的最佳用酶,然后在单因素的基础上以加酶量、液料比、酶解时间为自变量,DPPH自由基清除率为响应值,建立二次回归设计模型,经修正后得到最佳酶解条件为:加酶量为0.66%(w/w),液料比为10.2(v/w),酶解时间为380 min,在此条件下DPPH自由基清除率为89.80%,与模型预测值相吻合。另外,氨基酸分析表明与鸢乌贼蛋白相比,优化后的酶解产物中必需氨基酸含量提高至原蛋白的2.2倍,蛋氨酸、色氨酸等抗氧化性氨基酸分别提高至7.2、25.6倍。

Response surface methods were used to optimize the hydrolysis conditions of purpleback flying squid （Sthenoeuthis oualaniensis）for production of antioxidant peptides.Papain was the optimum enzyme selected by the single-factor tests.Based on the single-factor tests,enzyme amount,liquid-material ratio and hydrolysis time were determined by single-factor tests as independent variables, 1, 1-diphenyl-2-picrylhydrazyl （DPPH）radical scavenging activity as the response value. The model of quadratic regression design was established. The hydrolysis conditions were adjusted to that enzyme amount was 0.66% （w/w）, liquid-material ratio was 10.2（v/w）, hydrolysis time was 380 min. Under such conditions,1, 1 - diphenyl- 2- picrylhydrazyl （DPPH） radical scavenging activity was 89.80%.The result was consistent with the model prediction.In addition, amino acid analysis showed that compared with purpleback flying squid protein,the content of essential amino acids of enzymatic hydrolysis product was increased to 2.2 times,antioxidative amino acids such as methionine and tryptophan was increased to 7.2,25.6 times, respectively.