酶解糙刺参内脏团蛋白制备抗氧化肽的工艺研究

Optimization of enzymatic hydrolysis parameters for antioxidant capacity of peptide from sea cucumber(Stichopus horrens) viscera

为了能够有效利用糙刺参内脏蛋白,以水解度为主要指标,对酶解蛋白酶进行了筛选,并系统研究了酶解温度、p H、加酶量和水解时间等单因素对水解度的影响。在此基础上,利用响应面中心组合设计实验,对酶解工艺进行了优化,得到以温度、p H、加酶量及水解时间为因子的二次方程,通过方差分析和验证性实验得出,此二次方程能较好反应海参内脏团蛋白水解度的变化规律,得到最佳水解条件为:温度52.66℃,p H 6.71,加酶量2.5%(质量分数),酶解时间10 h,预测最高水解度为49.58%。最佳酶解条件下所得多肽74%以上分子质量小于500 Da。利用DPPH法测定最佳条件酶解所得多肽的抗氧化活性,3 g/L的抗氧化率达到了24.8%,利用ABTS法测定最佳条件酶解所得多肽的抗氧化活性,3 g/L的抗氧化率达到了0.255μmol/g。为海参内脏蛋白的利用提供了参考。

In order to increase the utilization value of sea source of peptides, protamex was selected for the hydrolysis cucumber （Stichopus horrens） viscera as a potential of sea cucumber viscera. One-factor-at-a-time method was used to investigate the effects of temperature, pH, enzyme dosage, and hydrolysis time on hydrolysis degree. Following these investigation, a quadratic regression model describing hydrolysis degree as a response to s tem- perature, pH, enzyme dosage, and hydrolysis time was set up using a Box-Behnken experimental design. As dem- onstrated by the results analysis of variance and validation experiments, the model could well indicate the hydroly- sis degree of sea cucumber viscera. The optimal hydrolysis conditions of temperature, pH, enzyme dosage, and hy- drolysis time were determined to be 52.66℃, 6.71, 2.5% （dry mass ratio） and 10 hour, respectively. Under these conditions, the hydrolysis degree of sea cucumber viscera is 49.58%. Under the best condition of enzymatic hy- drolysis, more than 74% of the peptides were less than 500 Da. The antioxidant activity of peptides was measured by DPPH method, showing that the radical scavenging activity is 24.8% when the concentration was about 3 mg/mL The antioxidant activity of peptides was measured by ABTS method, showing that the Vitamin C Equivalent Anti- oxidant Capacity is 0.255 μmol/g when the concentration was about 3 mg/mL.