摘要
植物内生菌株EJS-3发酵液经离心除菌,硫酸铵分级沉淀,Hiprep phenylFF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-PAGE)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。
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 SDS-PAGE analysis.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.
出处
《食品科学》
CAS
CSCD
北大核心
2010年第15期231-235,共5页
Food Science
基金
国家“863”计划项目(2008AA10Z309)
江苏省高新技术计划项目(BG2007335)
江苏省科技支撑计划项目(BE2008308)
关键词
植物内生菌
多黏类芽孢杆菌
纤溶酶
纯化
溶栓作用
endophytes
Paenibacillus polymyxa
fibrinolytic enzyme
purification
thrombolytic effect
