Molecular cloning and characterization of CMCase gene (celC) from Salmonella typhimurium UR.
The sequence coding for carboxymethylcellulase (CMCase, CelC) was isolated from the DNA of Salmonella typhimurium UR1. Comparison between the deduced amino acid sequence of CelC (368 amino acid residues, Molecular mass 41 kDa) and that of the previously published CMCase revealed that this enzyme belongs to the cellulase family 8 and D. The protein was overproduced in Escherichia coli using T7 expression system, and its activity was confirmed by CMC-SDS-PAGE. When the overexpressed CelC protein was tested on cellulose-type substrates, the recombinant protein is able to degrade cellulose-type substrates, such as CM-cellulose, xylan, avicel, lichenan, and laminarin. Optimal temperature and pH for enzyme activity were found to be 50 degrees C and pH 6.5, respectively.
Division of Biotechnology, Faculty of Natural Resources and Life Science, Dong-A University, Busan, 604-714.
Ju-Soon
JS
Yoo
Youn-Ju
YJ
Jung
Soo-Yeol
SY
Chung
Young-Choon
YC
Lee
Yong-Lark
YL
Choi
D002480:Cellulase / Q000235:genetics* / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015183:Restriction Mapping; D012486:Salmonella typhimurium / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D013379:Substrate Specificity
D002482:Cellulose; D002480:Cellulase; C019826:carboxymethylcellulase