酶解明胶制备抗氧化多肽清除DPPH·自由基的响应面优化

Response Surface Optimization of Enzymatic Hydrolyze Gelatin Preparing Antioxidant Peptide Clearance Rate of DPPH·free Radicals

在测量样品蛋白质含量的基础上,利用六种不同的蛋白酶水解猪皮明胶,以DPPH·自由基的清除率为因素,筛选最适的水解蛋白酶和水解条件。结果表明:酸性蛋白酶和木瓜蛋白酶水解物抗氧化功效最优,水解前使用缓冲液对原料进行预处理,水解中进行搅拌可以显著提高水解物的抗氧化效果。从生产成本考虑,建议工业化生产中使用木瓜蛋白酶进行水解。进一步采用木瓜蛋白酶在单因素试验基础上采用响应面分析方法优化酶水解条件,得到最佳酶解工艺条件为:pH6.40、温度60.00℃、酶料比0.03、时间4.06 h,在此条件下,木瓜蛋白酶水解产物对DPPH·的理论清除率为90.91%,试验数据为88.50%。

On the basis of measuring protein content of the sample, six different proteases used to hydrolyzate pig skin gelatin. The rate of DPPH radical scavenging was a filtering factor. The results showed that hydrolyzates of acid and papain protease had optimal antioxidant activity. Pretreating raw materials with buffer reagents and mixing can significantly improve antioxidant effect of hydrolysates. Considering the cost of production, hydrolysates of papain was recommended to the industrial production.Next, we used papain as catalyst to hydrolyze pig gelatin. The hydrolysis conditions were optimized by response surface methodology on the basis of single factor experiments. The optimized conditions were as follows： pH 6.40, hydrolysis temperature 60.00℃, enzymatic enzyme concentration 3% and hydrolysis time 4.06 h. Under this conditions, the DPPH.scavenging rate of the hydrolyzate was up to 90.91%. The experimental result was 88.50%.