尖吻蝮蛇蛇毒纤溶酶的分离纯化及氨基酸序列测定

Purification and determination amino acid sequence of fibrinolytic enzyme from Agkistrodon acutus venom

目的:从尖吻蝮蛇蛇毒中分离纯化出高纯度的纤溶酶并对其进行初步性质研究和氨基酸序列分析。方法:采用DEAE-Sepharose FF离子交换和两次Superdex75PG凝胶过滤,从尖吻蝮蛇蛇毒中纯化出可直接溶解纤维蛋白原的蛋白酶。结果:分离到的纤溶酶相对分子质量为25kD,HPLC分析纯度在97%以上,氨基酸序列分析其N末端为MSTEF QRYME IVVNHS MVKKY NGDSD KIKAW。结论:采用此工艺可从尖吻蛇毒中分离出高比活的纤溶酶,为进一步研究其药理作用奠定了基础。

Aim ：To isolate the fibrinolytic enzyme from Agkistodon acutus venom, and define its characters and the amino acid sequence of the product. Methods：The fibrinolytic enzyme was purified from the Agkistrodon acutus venom through DEAE-Sepharose FF ion-exchange chromatography and Superdex 75 PG gel column chromatography. Results： The molecular weight of the separated enzyme was 25 kD by means of SDS-PAGE and its purity is more than 97% estimated by HPLC. The N-terminal sequence of the product was confirmed as MSTEF QRYME IVVNHS MVKKY NGDOD KIKAW. Conclusion：The process is practicable for separation of pure fibrinolytic enzyme with high specific activity from Agkistrodon acutus venom. It lay a foundation for the further pharmacological study.