尖吻蝮蛇毒38k凝血酶样酶的分离纯化工艺

Purification Process of 38ku Thrombin-Like Enzyme from Agkistrodon acutus Venom

建立从尖吻蝮蛇毒中分离纯化38k凝血酶样酶的生产工艺,利用离子交换和凝胶过滤技术,中间品加入巯基乙醇处理后,再利用凝胶过滤技术除去性质相近的杂蛋白,获得的38k凝血酶样酶HPLC纯度达99.93%,比活力达4416U/mg,出血毒试验800U/ml无出血,所建立的分离纯化方法适合规模生产。

The thrombin-like enzyme isolated from Agkistrodon acutus venom by ion exchange and gel chromatography and the addition of ME, showed one band on SDS-PAGE with the relative molecular mass of the enzyme 38 ku, HPLC purity of 99. 3%. It has no hemorrhage by hypodermic injection 800U/ml to mice. TLE specific activity was 4416U/mg, and the output of TLE from one gram crystal venom of Agkistrod on acutus could be up to 10000U. 38ku TLE could be prepared by adopting this method, the method was suitable for large-scale production.