Journal
BIOCHEMICAL ENGINEERING JOURNAL
Volume 69, Issue -, Pages 204-210Publisher
ELSEVIER
DOI: 10.1016/j.bej.2012.09.011
Keywords
Cellulase; Co-expression; Escherichia coli; Enzyme production; Enzymes; Cellulose
Funding
- China National Natural Science Foundation of China [21176215/21176102]
- Outstanding Young Scholar of Zhejiang Province [R4110092]
- Program for Zhejiang Leading Team of ST Innovation [2011R50007]
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Usually, glucose production from cellulose hydrolysis involves synergistic actions by exoglucanase, endoglucanase and beta-glucosidase. To co-produce a whole cellulase system in Escherichia coli, a multi-promoter construct and a multi-cistronic construct harboring an exoglucanase (CbhA), an endoglucanase (CenA) and a beta-glucosidase (BGL) were built herein. Synergistic effect of three enzymes was also investigated. The results showed synergism of CenA and BGL played a critical role in degrading filter paper to glucose. Compared with monocistronic constructs, multi-promoter and multi-cistronic constructs harboring three enzymes showed significantly higher glucose yield. The specific activity of multi-promoter construct was 22.5-fold and 6.9-fold higher than CbhA and CenA, respectively and that of multi-cistronic construct was 33.6-fold and 10.3-fold higher than CbhA and CenA, respectively. In conclusion, a whole cellulase system could be co-produced in E. coli for converting cellulose to glucose. Moreover, the strategy could be easily applied to other cellulolytic enzymes, as well as for other multi-enzymatic systems. (C) 2012 Elsevier B.V. All rights reserved.
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