食品科学 ›› 2010, Vol. 31 ›› Issue (15 ): 231-235.doi: 10.7506/spkx1002-6630-201015050

• 生物工程 • 上一篇    下一篇

内生多黏类芽孢杆菌纤溶酶的纯化及其体外溶栓作用

吕凤霞,姚正颖,别小妹,赵海珍,王 煜,郭 瑶,陆兆新*   

  1. 南京农业大学食品科技学院
  • 收稿日期:2010-04-19 出版日期:2010-08-15 发布日期:2010-12-29
  • 通讯作者: 陆兆新* E-mail:fmb@njau.edu.cn
  • 基金资助:

    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)

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*   

  1. (College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China)
  • Received:2010-04-19 Online:2010-08-15 Published:2010-12-29
  • Contact: LU Zhao-xin* E-mail:fmb@njau.edu.cn

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 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-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。

关键词: 植物内生菌, 多黏类芽孢杆菌, 纤溶酶, 纯化, 溶栓作用

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, RESOURCEMQ 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.

Key words: endophytes, Paenibacillus polymyxa, fibrinolytic enzyme, purification, thrombolytic effect

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