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目的选用不同种类的水产原料用胰蛋白酶酶解,制备出具有抑制神经氨酸酶(NA)活性的多肽。方法选用中国毛虾、鲢鱼、鳀鱼、鳕鱼皮4种原料采用胰蛋白酶酶解,通过体外检测抑制NA活性和MDCK细胞感染H1N1病毒实验,得到能够抑制NA活性的多肽,考察了酶的种类、酶解时间、加酶量、料液比对活性的影响。结果与结论筛选出中国毛虾为原料,通过单因素实验确定酶解时间为5.5h,加酶量为2000U/g,料液比为1∶7(质量浓度)是胰蛋白酶酶解中国毛虾的最优条件。测得IC50值为1.32mg/mL。MDCK细胞感染H1N1病毒实验显示,浓度为250ug/mL的中国毛虾多肽的抑制效果较为明显。通过氨基酸分析推测多肽中高含量极性氨基酸能提高NA抑制活性。
Objective To select peptides from different kinds of aquatic raw materials for trypsin digestion and to inhibit the activity of neuraminidase (NA). Methods Four kinds of raw materials of Chinese shrimp, silver carp, bighead carp and cod skin were selected by trypsin digestion. The inhibitory activity of NA and the MDCK cell-infected H1N1 virus were tested in vitro to obtain the polypeptides capable of inhibiting NA activity. The enzyme types , Hydrolysis time, amount of enzyme, the ratio of material to liquid on the activity. RESULTS AND CONCLUSIONS The Chinese shrimp were screened out as raw materials. The enzymolysis time was 5.5h, the amount of enzyme was 2000U / g and the ratio of material to liquid was 1: 7 (mass concentration) by trypsinization. The optimal conditions. The IC50 value was found to be 1.32 mg / mL. MDCK cell infection H1N1 virus experiments showed that the concentration of 250ug / mL Chinese crude shrimp polypeptide inhibitory effect is more obvious. It is deduced by amino acid analysis that the high content of polar amino acids in the polypeptide can increase NA inhibitory activity.