一种新型蚯蚓纤溶酶的部分性质研究

Characterization of a novel earthworm fibrinolytic enzyme from Eisenia faetida

为获得一种高效的溶栓药物。从赤子爱胜蚓(Eiseniafoelida)中分离纯化得到了一种纤溶酶组分。用Lowry法测定蛋白质浓度,SDSPAGE鉴定纯度为98%,表观相对分子质量(Mr)为14850,纤维蛋白平板法测定其总纤溶活性为65.51×103mm2/mg,直接纤溶活性为15.61×103mm2/mg,间接纤溶活性为26.34×103mm2/mg。水解BAEE的米氏常数(Km)为1.82×105mol/L。水解ChromozymPL的米氏常数(Km)3.98×105mol/L,水解ChromozymtPA的米氏常数(Km)5.55×105mol/L活性,N端氨基酸序列测定的结果为VIGGTNAIPGEFPYQ。结果表明该纤溶酶分子量较小,间接活性较高,适宜作为一种新型的溶栓药物。

In order to obtain an effective drug for thrombolysis. A fibrinolytic enzyme was isolated and purified from the earthworm, Eisenia faetida. The protein concentration was determined by Lowry method. The purity and molecular weight was estimated by SDS-PAGE. The fibrinolytic activity was determined by Fibrin plate method. BAEE, Chromozym PL and Chromozym t-PA were used as specific substrate to obtain the Km.The sequence of 15 amino acids in N-terminus was determined by applied biosystem 491 protein sequencer. The results show that the purified enzyme is a new and effective drug for thrombolysis.