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目的自鳕鱼皮中制备流感病毒神经氨酸酶抑制活性肽。方法选用9种蛋白酶对鳕鱼皮胶原蛋白进行酶解,以神经氨酸酶抑制率为指标进行比较,选择最佳水解蛋白酶,用响应面法对酶解条件进行优化。结果与结论胃蛋白酶水解产物的抑制活性最高,且呈剂量依赖性,IC50值为8.86mg.mL-1。响应面分析确定最佳酶解条件为加酶量1600U.g-1,底物浓度22mg.mL-1,酶解时间6.8h。在此最佳条件下,胃蛋白酶水解产物IC50值为7.95mg.mL-1。抗H1N1流感病毒活性的细胞实验结果显示,10mg.mL-1的胃蛋白酶酶解物病毒抑制率可达66.6%。
Objective To prepare influenza virus neuraminidase inhibitory active peptides from cod skin. Methods Nine kinds of protease were used to hydrolyze cod skin collagen. Neuraminidase inhibition rate was used as an index to select the best hydrolytic protease. Response surface methodology was used to optimize the enzymatic hydrolysis conditions. RESULTS AND CONCLUSION Pepsin hydrolyzate had the highest inhibitory activity in a dose-dependent manner with an IC50 value of 8.86 mg.mL-1. Response surface analysis showed that the optimum conditions of enzymolysis were 1600U.g-1 enzyme concentration, 22mg.mL-1 substrate concentration and 6.8h hydrolysis time. Under these optimal conditions, the IC50 value of pepsin hydrolysates was 7.95 mg.mL-1. The experimental results of the cell activity against the H1N1 influenza virus showed that the inhibition rate of the pepsin hydrolyzate of 10 mg.mL-1 was up to 66.6%.