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目的:探讨从五步蛇毒中分离纯化凝血酶样酶的有效方法。方法:用Superdex 75、Superdex 30凝胶过滤色谱和DEAE-Sepharose Fast Flow阴离子交换色谱技术层析,SDS-PAGE分析纯度。结果:从五步蛇毒中分离纯化得到一个具有凝血活性的组分。SDS-PAGE显示为单一条带,相对分子质量约25 kDa。结论:用Superdex75、Superdex30凝胶过滤色谱和DEAE-Sepharose Fast Flow阴离子交换色谱技术可以很好分离纯化五步蛇毒中凝血酶样酶。
Objective: To explore an effective method for the isolation and purification of thrombin-like enzyme from five-step snake venom. Methods: The purity was determined by Superdex 75, Superdex 30 gel filtration chromatography and DEAE-Sepharose Fast Flow anion exchange chromatography. SDS-PAGE was used to analyze the purity. Results: A clotting-active component was isolated and purified from five-step snake venom. SDS-PAGE showed a single band, relative molecular mass of about 25 kDa. Conclusion: The thrombin-like enzyme of five-step snake venom can be well separated and purified by Superdex75, Superdex30 gel filtration chromatography and DEAE-Sepharose Fast Flow anion exchange chromatography.