Engineering of Clostridium phytofermentans Endoglucanase Cel5A for improved thermostability.
A family 5 glycoside hydrolase from Clostridium phytofermentans was cloned and engineered through a cellulase cell surface display system in Escherichia coli. The presence of cell surface anchoring, a cellulose binding module, or a His tag greatly influenced the activities of wild-type and mutant enzymes on soluble and solid cellulosic substrates, suggesting the high complexity of cellulase engineering. The best mutant had 92%, 36%, and 46% longer half-lives at 60 degrees C on carboxymethyl cellulose, regenerated amorphous cellulose, and Avicel, respectively.
Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Wenjin
W
Liu
Xiao-Zhou
XZ
Zhang
Zuoming
Z
Zhang
Y-H Percival
YH
Zhang
D002480:Cellulase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006358:Hot Temperature; D015202:Protein Engineering; D055550:Protein Stability; D013997:Time Factors
D002482:Cellulose; C407024:endoglucanase Cel5A; D002480:Cellulase