枯草杆菌溶栓酶的分离纯化研究

目的 :以枯草杆菌HL 986发酵液为原料 ,对枯草杆菌溶栓酶的分离纯化技术进行研究。方法 :采用磷酸钙吸附沉淀除去部分色素和杂蛋白 ,经超滤脱盐、CM 5 2离子交换分离、丙酮沉淀和SephadexG 75凝胶过滤处理 ,得到具有溶栓活性的酶。结果 :得到的溶栓酶在SDS PAGE上为单一谱带 ,其分子量约为 2 7kD ,比活为980 0U/mg,活性得率为 6 3.8%。结论 :以较高的得率获得了高纯度的枯草杆菌溶栓酶。

Purification of a Thrombolytic Enzyme from Bacillus subtilis HL -986

Purpose:The aim is to study purifying methods of a thrombolytic enzyme from the culture supernant of Bacillus subtilis HL-986.Methods :Some pigments and proteins were removed from the culture supernatant by C a 3(PO 4) 2 adsorption.Then the purified thrombolytic enzyme was obtained by sequential use of ultrofiltration with 10K hollow fiber membrane,ion-exchange c hromatography on CM-52,acetone precipitation,and gel-filtration on Sephadex G -75.Results:Specific activity of the enzyme was 9 800 IU/mg.The total activity recovery rate of enzyme was 63.8%.Its molecule weight is approxi mately 27kD and purified enzyme appears single band on SDS-PAGE.Conclusio n:Purified enzyme was obtained by the common methods and the total recover y rate was high.