裂褶菌（Schizophyllum commune）溶纖活性蛋白分離純化與其特性分析   全文下載

Purification and Characterization of a Novel Fibrinolytic Protease from Schizophyllum commune

裂褶菌（Schizophyllum commune）是一廣泛分布於世界各地的菇蕈種，有報告指出，裂褶菌菌絲體具有溶解纖維蛋白（fibrinolytic）之活性，因此目前被認為其有助於抗血栓之治療與應用。在本研究中，由人工液態發酵模式大量培養裂褶菌菌株，並利用橫掃式分子過濾系統以及快速液相層析技術（FPLC），分離純化醱酵溶液中的溶纖活性蛋白?，並針對此活性蛋白?進行特性分析。結果顯示，裂褶菌溶纖蛋白?之蛋白?單位酵素活性較原有培養液增加9.29倍，溶纖活性表現較市售人類溶纖蛋白?（plas-min）為高；且之，EDTA具有抑制其蛋白?活性的影響；於蛋白質特性表現：溶纖蛋白?分子量為21.32 kDa且具有單體蛋白質結構，也分別於pH 5.0與45°C狀態下具有最佳酵素活性；此外，由本實驗中亦發現鎂離子的作用可明顯提升溶纖蛋白?酵素活性近四倍。

Schizophyllum commune, a widely distributed medicinal mushroom, was found with fibrinolytic activity from its basidio-mycetes. The fibrinolytic activity of S. commune was to be beneficial for antithrombotic therapy. In this study, S. commune was cultured with fermentation technology, and protease purification was carried out by cross-flow filtration and fast performance liquid chromatography (FPLC) system. The specific activity of S. commune fibrinolytic protease increased 9.29-fold over the culture broth after purification. The fibrinolytic protease shows superior fibrinolytic activity than human plasmin and is inhibited by EDTA. Characterizations of this protease showed 21.32 kDa in molecular mass and monomeric form in protein structure. The optimal protease activity reveals at pH 5.0 and 45°C, and is enhanced by magnesium.