蚯蚓纤溶酶的分离纯化及性质鉴定

以赤子爱胜蚓为材料，经过硫酸铵盐析、DEAE-Sepharose CL-6B离子交换层析、Sephadex G-75凝胶过滤和制备电泳分离得到四种纤溶酶组分．经PAGE鉴定四种组分为单一组分；SDS-PAGE测定其分子质量分别为27、28、23和33ku；等电点均在4．0以下；经甲苯胺蓝染色后都出现糖蛋白的阳性反应；用苯酚一硫酸法测定其中性糖含量分别为8.4％、13.5％、9．1％、6．8％；纤维平板法测得四种组分的活性存在明显差异；四种组分都具有直接溶解纤维蛋白和激活纤溶酶原的双重作用．pH值对四种组分纤溶活性稳定性的影响各不相同．四种组分对温度的适应范围较广．

Purification and Characterization of Fibrinolytic Enzymes from Earthworm Eisenia foelide

Four components of fibrinolytic enzyme (EFE 1,EFE 2,EFE 3,EFE 4) were isolated and purified from earthworm Eisenia foelide . The procedure of purification included ammonium sulphate precipitation, ion exchange chromatography on DEAE Sepharose CL 6B, Sephadex G 75 gel filtration and preparative PAGE electropheresis. Four purified components which all showed fibrinolytic activities on fibrin plate were proved to be homogeneous in SDS PAGE electropheresis with molecular mass 27,28,23 and 33 ku respectively and their isoelectric point (pI) were below 4.0. They not only hydrolyzed fibrin directly, but also activated the plasminogen to plasmin. Some chemical and physical properties were characterized, they were confirmed to be glycoprotein. The amount of sugar contained in four components of EFE were about 9.1%, 13.5%, 8.4% and 6.8% respectively. Fibrinolytic activities of four components of EFE on fibrin plate were different. They were heat stable, pH stable and displayed a broad optimal pH range.