High-level expression, purification and characterization of a recombinant medium-temperature alpha-amylase from Bacillus subtilis.
Alpha-amylases are important industrial enzymes with a wide range of applications. Although medium-temperature alpha amylase (AmyE) has some practical advantages, its low yield has limited its applications. When an amyE gene from Bacillus subtilis BF768 was cloned into vector pWB980 and over-expressed in B. subtilis WB600, high activities (723 U ml(-1)) of secreted AmyE were produced. Recombinant AmyE was purified to a specific activity of 36 U mg(-1) having optimal activity at pH 6.0 and 60 degrees C.
Liu
Yihan
Y
Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin, 300457, People's Republic of China.
Lu
Fuping
F
Chen
Guanqun
G
Snyder
Crystal L
CL
Sun
Jing
J
Li
Yu
Y
Wang
Jianling
J
Xiao
Jing
J
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D013696:Temperature; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D000516:alpha-Amylases