葡萄籽蛋白的碱性蛋白酶酶解工艺及其产物抗氧化活性研究

Study on Enzymatic Hydrolysis of Grape Seed Protein by Alcalase and Antioxidant Activities of Its Hydrolysate

采用响应曲面法对葡萄籽蛋白的碱性蛋白酶酶解工艺进行优化,并研究此酶解物的抗氧化活性。在单因素试验的基础上,选择pH、酶解温度和酶解时间进行3因素3水平的Box-Behnken试验设计,采用响应曲面法(RSM)分析3个因素对响应值的影响。结果表明:最佳酶解工艺参数为pH 8.2,酶解温度50.4℃和酶解时间2.71h,在此条件下水解度为24.66%;葡萄籽蛋白碱性蛋白酶酶解物在质量浓度112.5～562.5μg/mL和180～900μg/mL范围内,其对DPPH自由基和羟基自由基的清除率分别为16.96%39.02%和20.79%～86.73%,且都存在明显的量效关系。

To optimizethe enzymatic hydrolysis process for grape seed protein with alcalase by response surface methodology and explore antioxidant activities of its hydrolysate. Based on single factor experiments, a three-factor and three-level Box-Behnken design coupled with response surface analysis was employed to probe the effects of pH value, enzymolysis temperature and enzymolysis time on the degree of hydrolysis. It was found that the best extraction conditions were as follows： pH 8.2, enzymolysis temperature 50.4 ~C with enzymolysis time 2.71 h, and that the degree of hydrolysis was 24.66% under such conditions. When the concentrations were 112.5-562.5 ~tg/mL and 180~900 Ixg/ mL, the clearance rates of DPPH and .OH free radical were 16.96%-39.02% and 20.79%-86.73% individually, and the scavenging activity was correlated with the concentrations.