多指标优化珠蚌抗氧化肽制备工艺

Multi-index optimization of preparation parameters for antioxidant peptide from freshwater mussel

采用取珠后珠蚌肉为原料,选用木瓜蛋白酶进行水解。以DPPH自由基清除率、.OH清除率及蛋白水解度作为评价指标,研究珠蚌抗氧化活性肽的制备工艺。在单因素基础上,通过三个指标的综合考虑,设计L18(37)多指标正交试验,对制备工艺进行优化,得出最佳工艺条件。结果表明,最佳条件为:酶解温度60℃、酶解时间5h、料液比3:10(w/v)、酶解pH6.5、加酶量6000U/g,制得的多肽DPPH自由基清除率85.5%,.OH清除率50.7%,水解度23.09%;此时,DPPH自由基清除率EC50为0.4mg/mL,.OH清除率EC50为0.7mg/mL。

The protein taken off the freshwater mussel was hydrolyzed by pepsin. The optimal hydrolysis parameters of strongest antioxidant capacity of peptide were obtained using multi-index orthogonal experiment. The effects of reaction temperature, hydrolysis time, solid/liquid ratio, pH value and enzyme concentration on the scavenging rate of free radicals DPPH, and .OH of the peptide,and the degree of hydrolysis of protein were well fitted to a high determination coefficients. The hydrolysate with optimal antioxidant activity was predicted to be obtained at .. temperature 60℃,hydrolysis time 5h,solid/liquid ratio 3：10（w/v）,pH value 6.5,and enzyme concentration 6000U/g. Under these conditions,the scavenging rate on DPPH. was 85.5%,and the scavenging rate on -OH reached 50.7%,the degree of hydrolysis was 23.09%. Under this condition,the concentration for 50% of maximal effect （ECs0） of mussel antioxidative peptide on DPPH. and .OH radicals were 0.4mg/mL and 0.7mg/mL.