响应面法优化糙刺参内脏团制备抗氧化肽的酶解工艺

Screening for optimal enzyme for preparation of the visceral of stichopus horrens protein hydrolysate with antixodiant activities and process optimization by response surface methodology

以加工糙刺参的副产物内脏团为原料,选用碱性蛋白酶、复合蛋白酶、风味蛋白酶、木瓜蛋白酶和中性蛋白酶分别对其进行酶解,采用ABTS法对酶解产物进行抗氧化活性分析,筛选出酶解产物具有较强抗氧化活性的木瓜蛋白酶作为最适用酶,然后通过响应面法对酶解工艺条件进行了优化。结果表明:木瓜蛋白酶酶解糙刺参内脏团的最佳酶解参数为:温度为50.19℃,pH为8.0,加酶量为2.5%,酶解时间为9.0 h,此时3 mg/mL酶解物的还原能力(ABTS法)为0.2533μmol/g,实验验证结果为0.2552μmol/g;响应面法预测值与实验验证结果吻合性良好。此外,最佳酶解条件下所得酶解产物的氨基酸含量丰富,谷氨酸含量较多,分子量在2000 u以下的组分占总量的99.52%。

In the present study, the protein hydrolysates of Stichopus horrens viscera were prepared with alcalase, protamex, flavourzyme, neutrase and papain, respectively. Their Vc equivalent antioxidant capacity（VcEAC） was comparatively determined by the ABTS method, with the result that hydrolyzing Stichopus horrens visceral protein by papain to a hydrolysate with better VcEAC. Four main process conditions for the hydrolysis of Stichopus horrens visceral protein by protease, including temperature, pH, enzyme/substrate ratio and hydrolysis time were further optimized using response surface methodology（RSM）. The optimum technological parameter for preparing antioxidant peptide with papain was obtained as： pH 8.0 at 50.19 ℃, enzyme/substrate ratio 2.5% and the hydrolysis time 9.0 h, and the VcEAC of 3 mg/mL hydrolysates was 0.2533 μmol/g, the experimental result was 0.2552 μmol/g. The experimental results were good agreement with the predicted value. Furthermore, glutamic was the main amino acids in the enzymatic hydrolysis product, the molecular weight of under 2000 u components account for 99.52% of the total enzymatic hydrolysis product.