黑眉蝮蛇蛇毒纤溶酶的纯化及其性质研究

Isolation and characterization of fibrinolytic enzyme from Gloydius saxatilis snake venom

目的从黑眉蝮蛇蛇毒中纯化出无出血毒的纤溶酶。方法用DEAE-Sepharose FF阴离子交换树脂、CM-sepharose FF阳离子交换树脂和Sephacryl S-100凝胶过滤树脂三步分离方法,从黑眉蝮蛇蛇毒中纯化出一种纤溶酶活性成分。结果经SDS-聚丙烯酰胺凝胶电泳检测为单一蛋白质,相对分子质量为31 800,小鼠皮下注射无出血反应。该酶与纤维蛋白原保温15 min能迅速水解Bβ(β)链,随后缓慢水解Aα(α)链,而对γ(γ-γ)链无影响。结论此工艺可以快速纯化黑眉蝮蛇蛇毒纤溶酶。

Purpose To purify fibrinolytic enzyme of no hemorrhage from Gloydius saxatilis snake venom. Methods Fibrinolytic enzyme was purified from the venom of Gloydius saxatilis by the DEAE Sepharose FF ion exchange, CM-sepharose FF ion exchange and Sephacryl S-100 gel filtration column chromatography. Results The purified fibrinolytic enzyme was proved to be homogenous by SDS-PAGE gel electrophoresis, its molecular weight was about 31 800, and there was no hemorrhage by subcutaneous injection to mouse. The fibrinolytic enzyme rapidly degraded the fibrinogen Bβ （β） chain in 15mins, and subsequently hydrolyzed the Aα（α） chain after 2 h incubation, but it had no effect on γ（γ-γ） chain. Conclusion The process can be used to purify rapidly the fibrinolytic enzyme from Gloydius saxatilis snake venom.