自鳕鱼皮中制备流感病毒神经氨酸酶抑制活性肽

Preparation of active peptide from cod skins with inhibitory activity on influenza virus neuraminidase

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 optimize the preparation technology of active peptide from cod skins with inhibitory activity on influenza virus neuraminidase（NA）.Methods The collagen from cod skins was hydrolysed by nine proteases and the one was selected at the target of neuraminidase finhibition ratio.Three main factors were studied by response surface methodology（RSM）.Results and Conclusion Among the 9 kinds of proteases,the NA inhibition ratio of pepsin hydrolysates was the highest and showed a dose-dependent manner with it’s IC50 8.86 mg·mL^-1.The optimal enzymolysis conditions were： enzyme dosage 1600 U·g-1,substrate concentration 22 mg·mL^-1,hydrolyzing time 6.8 h,under which IC50 of NA inhibition ratio was 7.95 mg·mL^-1.The anti-virus rate was 66.6% on MDCK cells infected by H1N1 under the concentration of 10 mg·mL^-1.