中文摘要：

【目的】确立蛹拟青霉深层培养液中高纯度、高纤溶活性纤溶酶的分离纯化方法并测定其酶学性质。【方法】采用硫酸铵盐析、Sephadex G-25凝胶色谱、Phenyl-Sepharose HP疏水相互作用色谱、CM-Sepharose FF弱阳离子交换色谱和Superdex 75凝胶色谱对蛹拟青霉纤溶酶进行分离。用Lowry法测定蛋白质浓度, 纤维蛋白平板法测定其纤溶活性, SDS-PAGE鉴定其纯度并确定其分子量, IEF法测定其等电点。【结果】研究发现, 以蔗糖和豆饼为培养基主要基质时, 蛹拟青霉深层培养可以产生至少两种纤溶酶。提纯后的纤溶酶Ⅱ比活力达到800.46 U/mg, 总纯化倍数为30.07倍。纤溶酶Ⅱ的相对分子量和等电点分别为32 kD和9.3±0.2。纤溶酶Ⅱ是一种糖蛋白, 总含糖量为0.98% （w/v）。该酶可以顺次降解人血纤维蛋白（原）的α、β和γ链。其最适作用pH及温度分别为7.4和41 °C。Aprotinine与PMSF对该纤溶酶的活性完全抑制, 推测此纤溶酶可能是一种丝氨酸蛋白酶。【结论】单一的高纤溶活性纤溶酶的获得和酶学性质的确定，为该酶开发成为新型溶栓药物提供了理论依据。

英文摘要：

[Objective] Establish purification methods of fibrinolytic enzyme with high purity and fibrinolytic activity from Paecilomyces militaris submerged culture liquid and determine its characterization. [Methods] The fibrinolytic enzyme Ⅱ of Paecilomyces militaris was purified by ammonium sulfate precipitation, Sephadex G-25 gel filtration chromatography, Phenyl-Sepharose HP hydrophobic interaction chromatography, CM-Sepharose FF ion exchange chromatography and Superdex 75 gel filtration chromatography. The protein concentration of enzyme Ⅱwas determined by Lowry method. The fibrinolytic activity was determined by Fibrin plate method. The purity and molecular weight was estimated by SDS-PAGE. The isoelectric point was estimated by isoelectric focusing electrophoresis. [Results] We found that Paecilomyces militaris produced at least two kinds of fibrinolytic enzymes by submerged cultivation, sucrose and bean cake as major materials. The specific activity of purified fibrinolytic enzyme Ⅱ was 800.46 U/mg and the purification protocol resulted in 30.07-folds purification of the enzyme Ⅱ. The molecular weight and isoelectric point of the enzyme Ⅱ was 32 kD and 9.3±0.2, respectively. The enzyme Ⅱwas a glycoprotein, the total sugar content of it was 0.98% （w/v）. The enzyme Ⅱ could degrade α, β and γ chains of human fibrinogen. The optimal pH and temperature of the enzyme were 7.4 and 41°C. The fibrinolytic activity was completely inhabited by Aprotinine and PMSF, which indicated that it might be a serine protease. [Conclution] The acquisition of enzyme with single and high fibrinolytic activity and the determination of its characterization, which provides theoretical basis for the enzyme to become a new thrombolysis drug.