响应面法优化草菇抗氧化肽的酶法制备工艺

Optimization of Enzymatic Preparation Technology of Antioxidant Peptide from Volvariella volvacea by Response Surface Methodology

以草菇为原料提取蛋白质,蛋白酶酶解蛋白制备抗氧化肽。以DPPH自由基清除率为指标,在单因素实验基础上,结合响应面法优化草菇抗氧化肽的提取工艺。通过超滤分离纯化获得不同分子量的肽段,采用DPPH自由基清除率、Fe^(2+)螯合率和还原力法测定超滤组分的抗氧化活性。结果表明:中性蛋白酶为最优酶解蛋白酶,最佳酶解工艺条件为酶解时间3.70 h,加酶量3.81%,底物质量浓度3.11 g/100 mL,在此条件下,酶解产物的DPPH自由基清除率为69.85%±2.52%。通过超滤分级制备所得分子量最小的肽段F1(<3 kDa)具有最高的抗氧化活性,其DPPH自由基清除率、Fe螯合率和还原力分别为78.81%±1.56%、91.05%±1.65%、0.47±0.02。草菇抗氧化肽可作为潜在的天然抗氧化剂来源得到开发利用。

The antioxidant peptides were prepared from Volvariella volvacea protein by protease hydrolysis.Based on the single-factor test,the extraction conditions of antioxidant peptides from V.volvacea protein were optimized using response surface methodology.At the same time,peptides with different retention molecular weights were isolated by ultrafitration,and their antioxidant activity were detected through 2,2-dipheny1-1-picrylhydrazyl(DPPH)radical scavenging activity,Fe^(2+)chelating ability and reducing power.The results showed that the optimum enzyme for enzymatic hydrolysis was neutral protease,and the optimum process for antioxidant peptide was as follows:The enzymatic hydrolysis time was 3.70 h,the enzyme dosage was 3.81%,and the substrate mass concentration was 3.11 g/100 mL.Under the optimal enzymalysis conditions,DPPH free radical scavenging rate of the optimal hydrolysis products was 69.85%±2.52%.The F1 fraction with the lowest molecular weight(<3 kDa)exhibited the highest antioxidant activity.DPPH free radical scavenging rate,Fechelating ability and reducing power of F1 were 78.81%±1.56%,91.05%±1.65%,0.47±0.02.The antioxidant peptides from V.volvacea protein can be explored as a potential natural antioxidant.