广东菲牛蛭中有效抗凝物质的分离纯化

Purification of Anticoagulation Protein from the Guangdong Hirudinaria Manillensis

目的从广东菲牛蛭鲜体中分离纯化有效抗凝物质,并对其生化性质进行测定。方法用水提醇沉、离子交换、反相层析法等分离纯化广东菲牛蛭中的抗凝物质,用SDS-聚丙烯酰胺凝胶电泳测定其蛋白的分子质量,并运用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS/MS)进行分析。结果从广东菲牛蛭中分离出得到一种电泳纯的活性多肽,其相对分子质量为19.2kDa。所得菲牛蛭抗凝活性多肽每克蛋白所含单位抗凝活性≥2548000ATU,活性回收率约为53%。结论从广东菲牛蛭鲜体中分离出的活性抗凝多肽对凝血酶具有极强的抑制作用,开发应用前景广阔。

Objective To purify anticoagulation protein from the Guangdong ttirudinaria manillensis and to determine the characterization. Methods The anticoagulation protein was purified by water extract and alcohol precipitate and column chromatography on DEAE - Sepharose F.F. and Reverse - phase chromatography. The molecular weight was calculated Using SDS - PAGE and analysis was taken by the MALDI - TOF - MS/MS. Results The purified protein showed a single band by SDS - PAGE with a molecular weight of 19.2kDa. The specific activity of per gram purified protein from the Guangdong Hirudinaria manillensis was 2548000ATU, yield 53%. Conclusion The anticoagulation protein was purified from the Guangdong Hirudinaria maniUensis and showed the strong thrombin inhibitor.