中药蒲黄纤溶蛋白的分离纯化及部分性质

用DEAE Sepharose Fast Flow离子交换层析从中草药蒲黄中分离纯化一种纤维蛋白溶解酶,用纤维蛋白板检测分离后组分的纤溶蛋白活性,天然电泳检测分离效果,分离后得到了比活力为25.29的单一组分,SDS电泳确定该组分的表观分子量为31 000 D,同时比较了不同蛋白酶抑制剂对该组分的抑制作用。结果表明:该组分的纤维蛋白水解酶活性被PMSF、亮抑肽素、抑肽酶、大豆胰蛋白酶抑制剂以及苯甲脒强烈抑制,表明该粗提物中的蛋白酶属于丝氨酸蛋白酶家族。

Purification and characterization of fibrinolytic enzyme from Typha angustifolia L.

A fibrinolytic enzyme was isolated from a traditional Chinese medical named Typha angustifolia L.by a DEAE Sepharose Fast Flow column.The fibrinolysis activity of each isolated component was determined through fibrillarin-plate,and the purity of each component was determined by native protein electrophoresis.One purified single protein was obtained, whose specific activity was 25.29 and the molecular weight was 31 000 D confirmed by SDS protein electrophoresis.At the same time with inhibiting activities of various inhibitors on this protein were compared.The results indicated that it may be a serine family proteases,because the activity of the protein was inhibited powerfully by PMSF,Leupeptin,aprotinin,soybean trypsin inhibitor. This protein's activity was strong and the content was high and the molecular weight was suitable to molecular experiment.