榆干离褶伞溶栓酶的纯化及酶学性质研究

Purification of Fibrinolytic Enzyme from Cultured Mycelia of Lyophyllum ulmarium

采用离子交换层析、凝胶过滤层析及快速蛋白液相色谱等蛋白质分离纯化技术,从榆干离褶伞菌丝体中纯化一种溶栓酶,并探讨各种因子对榆干离褶伞溶栓酶活性的影响,观察溶栓酶对纤维蛋白及纤维蛋白原的水解特征和酰胺水解活性。实验结果表明,其分子质量约为50 ku,该酶的最适pH和最适温度为pH 6.0,35℃,并对tPA底物S-2288最敏感。Mn^(2+)和Mg^(2+)激活酶活性,而Cu^(2+),EDTA对酶活有抑制作用。该酶不仅直接水解纤维蛋白,还可以水解纤维蛋白原。榆干离褶伞很有潜力成为治疗血栓的新的药物来源。

A Fibrinolytic enzyme was purified from the cultured mycelia of Lyophyllurn ulrnarium by ion exchange chromatography followed by gel filtration and fast protein liquid chromatography. The factors affected on the fibrinolytic activity of fibrinolytic enzyme from the cultured mycelia of Lyophyllurn ulrnariurn, and the fibrinolytic specialty on fibrin and fibrinogen and amidolytic activity were examined was studied. The apparent molecular mass of purified enzyme was estimated to be 50 ku. Its optimal pH and temperature value were 6.0 and 35℃, respectively, and it was found to exhibit a higher specificity for the substrate S-2288 for tPA. Enzyme activity was enhanced by Mn^2＋ and Mg^2＋ , but inhibited by the addition of Cu^2＋ and EDTA. The mycelia of Lyophyllurn ulrnariurn may thus represent a potential source of new therapeutic agents to treat thrombosis.