响应面法优化复合酶酶解制备可口革囊星虫胶原蛋白抗氧化肽工艺研究

以可口革囊星虫为原料提取胶原蛋白，以总抗氧化能力和超氧阴离子清除率为考察指标，通过单因素实验和响应面法优化复合酶酶解制备可口革囊星虫胶原蛋白抗氧化肽工艺条件。结果表明，选用酶筛选试验中多肽抗氧化活性较高的木瓜蛋白酶、中性蛋白酶、胃蛋白酶和酸性蛋白酶进行单酶和复合酶试验，最佳方案确定为木瓜蛋白酶与中性蛋白酶复合酶解。在单因素实验基础上确定复合酶添加量（U/g）、酶解温度（℃）、酶解pH、酶解时间（h）为自变量，通过响应面法优化并参考实际因素，确定复合酶酶解的最优条件是:复合酶添加量8135 U/g、酶解温度51.6 ℃、酶解pH6.4、酶解时间4.2 h，在此条件下酶解制备的胶原肽总抗氧化能力为(1.333±0.021）μmol/mL，超氧阴离子清除率为78.75%±0.94%。

Optimization of the Preparation of Collagen Antioxidant Peptides from Phascolosoma esculenta by Compound Enzymatic Hydrolysis by Response Surface Methodology

In this study, collagen was extracted from Phascolosoma esculenta. The hydrolysis conditions for preparing the collagen antioxidant peptides of Phascolosoma esculenta was optimized by single factor test and response surface methodology with the total antioxidant capacity and superoxide anion clearance rate as inspection indicators. In the enzyme screening test, papain, neutral protease, pepsin and acid protease, which obtained polypeptides with higher antioxidant activity were selected for single and compound enzymatic hydrolysis tests. As a result, the compound enzyme hydrolysis of papain and neutral protease was determined as the best solution. On the basis of single factor test results, the compound enzyme addition amount (U/g), enzymatic hydrolysis temperature (℃), enzymatic hydrolysis pH, enzymatic hydrolysis time (h) were determined as independent variables. According to response surface analysis combining with actual factors, the optimal enzymatic hydrolysis conditions were determined as follows: Compound enzyme addition amount was 8135 U/g, enzymatic hydrolysis temperature was 51.6 ℃, enzymatic hydrolysis pH was 6.4, enzymatic hydrolysis time was 4.2 h. Under these conditions, the total antioxidant capacity of the collagen peptide was (1.333±0.021) μmol/mL, and the superoxide anion clearance rate was 78.75%±0.94%.