尖吻蝮蛇毒中一种新类凝血酶的分离纯化

Purification of a Novel Thrombin-Like Enzyme from Agkistrodon acutus Venom

用阴离子交换色谱和凝胶过滤色谱技术,从尖吻蝮蛇毒中纯化得到一个具有凝血活性的组分。经Superdex75HR10/30预装柱检测,纯度达99.91%。SDS-PAGE显示为单一条带,还原、非还原条件下分子量分别为59.25k、52.58k。该组分的凝血酶比活为41.5u/mg,精氨酸酯酶比活为14.29u/mg,但不激活血浆凝血因子ⅩⅢ。EDTA不能抑制其凝血活性,而苯甲磺酰氟则产生不可逆抑制作用。结果表明该酶是一种新尖吻蝮蛇毒类凝血酶。

A component with the clotting activity was isolated in 99.91% purity from Agkistrodon acutus venom by anion-exchange chromatography and gel filtration, The enzyme had a single band on SDS-PAGE, with Mr of 59.25k and 52.58k under reduced and non-reduced conditions. The clotting specific activity of the enzyme was 41.5u/mg on bovine plasma and the specific activity of arginine esterase was 14.29u/mg on BAEE. The enzyme did not activate coagulation factor ⅩⅢ, its activity to clot fibrinogen was inhibited irreversible by PMSF, but not by EDTA. The above results showed that the enzyme was a novel thrombin-like enzyme.