Bacillus subtilis Z-3菌株纤溶酶的分离纯化研究

Separation and purification of fibrinolytic enzyme from Bacillus subtilis Z-3

为获取微生物源溶栓药物,从土壤中获得产纤溶酶活性较高的1株枯草芽孢杆菌（Bacillus subtilis）Z-3。从Z-3菌株的发酵液中提取纤溶酶,利用DEAE-Sepharose Fast Flow阴离子交换层析和聚丙烯酰凝胶电泳等方法,对纤溶酶进行分离纯化。分离纯化得到1个单一组分（BSFE-1）,SDS-PAGE电泳检测为单一条带,BSFE-1表观分子量为48 kDa,等电点为10.5,属丝氨酸蛋白酶类,纤维蛋白平板法检测纤溶酶活性BSFE-1的比活力为4 494.099 U/mg,BSFE-1在4~50℃和pH 4~11之间活性较稳定。

The activity of a fibrinolytic enzyme produced from Bacillus subtilis Z-3 which was obtained from soil was examined by using fibrin plate method and its isoelectric point was determined via isoelectric focusing electrophoresis.By applying the DEAE-Sepharose Fast Flow anion exchange and polyacrylamide gel electrophoresis approaches,a single protein fraction（BSFE-1） with fibrinolytic activity was separated and purified from the culture of Bacillus subtilis Z-3.The SDS-PAGE result showed that BSFE-1 had only a single band with a relative molecular weight of 48 kDa and isoelectric point of 10.5.On fibrin plate,the specific activity of BSFE-1 with activity of serine proteins enzymes reached 4 494.099 U/mg.The fibrinolytic activity of BSFE-1 was stable at 4-40 ℃ and at pH 4-11.The separation of the fibrinolytic enzyme with high efficiency,specificity,security and few side effects will lay the foundation for the investigation of thrombolytic drugs.