内生多黏类芽孢杆菌纤溶酶的纯化及其体外溶栓作用-河南农业科学.PDF

※生物工程 食品科学 2010, Vol. 31, No. 15 231 吕凤霞，姚正颖，别小妹，赵海珍，王 煜，郭 瑶，陆兆新 * (南京农业大学食品科技学院，江苏 南京 210095) ：植物内生菌株 EJS-3 发酵液经离心除菌，硫酸铵分级沉淀，Hiprep phenyl FF 疏水层析，RESOURCEM Q 离子交换层析和 Sephacryl S-300HR 凝胶过滤，获得电泳纯的多黏芽孢杆菌纤溶酶(PPFE-I) ，聚丙烯酰胺凝胶电泳 (SDS)测定其分子质量为 63kD，高效液相色谱法(HPLC)鉴定其纯度为 94.1% 。每升发酵液中可获得 1.6mg 活 性蛋白，每毫克蛋白活力达 2096IU ，纯度提高了 14.5 倍，回收率为 3.3% 。纯化后的纤溶酶PPFE-I 具有明显的体 外溶栓作用。 ：植物内生菌；多黏类芽孢杆菌；纤溶酶；纯化；溶栓作用 Purification and Thrombolytic Effect in vitro of a Novel Fibrinolytic Enzyme Produced by Endophytic Bacterium Paenibacillus polymyxa EJS-3 . . LU Feng-xia ，YAO Zheng-ying ，BIE Xiao-mei ，ZHAO Hai-zhen ，WANG Yu ，GUO Yao ，LU Zhao-xin* (College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China) Abstract ：Endophytes are a very important resource of microorganisms with potential application values. A novel fibrinolytic enzyme (PPFE-I) was purified from the fermentation supernatant of endophytic bacterium Paenibacillus polymyxa EJS-3 by the following procedure including ammonium sulfate precipitation, Hiprep phenyl FF hydrophobic chromatography, RESOURCEM Q ion exchange chromatography and Sephacryl S-300HR gel filtration. The apparent molecular mass of PPFE-I was estimated to be 63 kDa by SDSanalysis. The purity of PPFE-I was 94.1% determined by HPLC. Using the above procedure, 16 mg of active proteins were obtained from 1 L of the fermentation supernatant of Paenibacillus polymyxa EJS-3 and a 14.5-fold increase in the specific activity of PPFE-I was achieved and the activity recovery was 3.3%. Meanwhile, the final purified product had good thrombolytic activity in vitro. Key words ：endophytes ；Paenibacillus polymyxa ；fibrinolytic e