海参蒸煮废液多肽提取响应面优化及抗氧化研究

Optimization of Preparation Procedure and Antioxidant Activity of Polypeptide from Sea Cucumber Cooking Waste Liquid

以海参蒸煮废液为原料,采用蛋白酶酶解法提取废液中的海参多肽,筛选出水解海参蛋白的最适蛋白酶。通过单因素试验,研究了加酶量、酶解pH、酶解温度和酶解时间四个因素对水解度的影响,并通过Box-Benhnken中心组合试验及响应面分析法对酶解条件进行优化。结果表明,中性蛋白酶水解效果最好,其最优酶解工艺条件为:酶解时间4 h,加酶量3.85%,温度45℃, pH 7.0。在此条件下海参蛋白水解度为44.5%。制得的海参多肽对羟自由基和超氧自由基具有较强的清除能力, IC_(50)值依次为6.901和8.804 mg/mL。

In this experiment, the sea cucumber cooking waste liquid was used as raw material, and the sea cucumber polypeptide in the waste liquid was extracted by using enzymatic hydrolysis method with the optimal protease（neutral protease）. On the basis of single-factor test, the mathematical regression model was established about the dependent variable（degree of hydrolysis） and in dependent variables（pH, hydrolysis temperature and dosage of neutral protease） through BoxBenhnken center composite design and response surface methodology. The results indicated that the optimal extraction conditions were hydrolysis time 4 h, dosage of neutral protease 3.85%, hydrolysis temperature 45 ℃, and pH 7.0. Under these conditions, the degree of hydrolysis was 44.5%. The sea cucumber polypeptide had strong scavenging capacity for hydroxyl radical and superoxide radical, and the IC50 value was 6.901 mg/mL and 8.804 mg/mL, respectively.