[Expression, purification and enzymatic characterization of Bacillus polymyxa beta-glucosidase gene( bglA ) in Escherichia coli].
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.
Zhao
Yun
Y
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Liu
Wei-Feng
WF
Mao
Ai-Jun
AJ
Jiang
Ning
N
Dong
Zhi-Yang
ZY
D001407:Bacillus / Q000201:enzymology*; D004926:Escherichia coli / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D001617:beta-Glucosidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins; D001617:beta-Glucosidase