海南菲牛蛭抗凝血酶成分的提取与SDS-PAGE电泳测定

Extraction and SDS-PAGE Electrophoresis of Anticoagulant Components from Poecilobdella manillensis Lesson in Hainan

应用凝血酶滴定法,比较不同提取方法的提取效率和不同相对分子质量的超滤膜对菲牛蛭抗凝血酶活性成分的分离作用,并用SDS-PAGE电泳测定分离得到抗凝活性成分的相对分子质量。结果表明:盐提取法更优于水提法、水提醇沉法和丙酮法;用盐提法对菲牛蛭头部提取的抗凝成分活性达1 811.11 U·g^-1,选择10×10^3和30×10^3的超滤膜进行再次超滤得抗凝成分相对分子质量约为15.96×10^3。

To study the anticoagulant activity and purify the anticoagulation components from P.manillensis in Hainan.Poecilobdella manillensis Lession collected in Hainan was extracted with different extraction methods and purified with molecular weight ultra membranes to determine its anticoagulant components and activities by using the thrombin titration method.The relative molecular weight of the anticoagulant components was calculated by using the SDS-PAGE test.The results showed that the sodium chloride solution was better than the water extraction,the water extraction and alcohol precipitation,and the acetone extract in extraction of anticoagulant components.The anticoagulant activity of the extract from the head of P.manillensis extracted by the sodium chloride solution was 1 811.11 U·g^-1,the highest among the other extraction methods.The ultra-membranes with molecular weights of 10×10^3 and 30×10^3 were selected for purification,and the molecular weight of the purified protein was 15.96×10^3.