Expression, purification and enzymatic characterization of Bacillus polymyxa beta-glucosidase gene( bglA ) in Escherichia coli.
- Author:
Yun ZHAO
1
;
Wei-Feng LIU
;
Ai-Jun MAO
;
Ning JIANG
;
Zhi-Yang DONG
Author Information
1. Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
- Publication Type:Journal Article
- MeSH:
Bacillus;
enzymology;
Escherichia coli;
genetics;
Plasmids;
Recombinant Proteins;
biosynthesis;
isolation & purification;
beta-Glucosidase;
genetics;
isolation & purification;
metabolism
- From:
Chinese Journal of Biotechnology
2004;20(5):741-744
- CountryChina
- Language:Chinese
-
Abstract:
The beta-glucosidase encoding gene bglA was cloned from Bacillus polymyxa 1.794. The bglA gene was inserted in expression vector pET28a(+) and transformed into Escherichia coli BL21 (DE3), finally the recombinant strain BL1979 was obtained. Induced by IPTG, the expression P-glucosidase activity reached to 24.7 IU/mL. The optimum temperature and optimum pH of the recombinant expression P-glucosidase in BL1979 were 37 degrees C and 7.0 respectively,the purity can reach to 92.7%. Analysis of the fusion protein by nondenaturing gradient gel electrophoresis, we found the fusion protein exists in dimmer, tetramer,hexamer and octamer, they all have hydrolase activity.
