Abstract
To engineer a multifunctional xylan-degrading enzyme, a chimera was created by fusing the xylanase domain of the Clostridium thermocellum xylanase (xynZ) and a dual functional arabinofuranosidase/xylosidase (DeAFc; from a compost starter mixture) through a flexible peptide linker. The xylanase domain of xynZ possesses previously unreported endoglucanase activity. The chimera, possessing the activities of xylanase, endoglucanase, arabinofuranosidase and xylosidase, was expressed in E. coli and purified. The chimera closely resembled the parental enzymes in pH, temperature optima and kinetics, and was more active than the parental enzyme mixture in the hydrolysis of natural xylans and corn stover.
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Acknowledgements
We thank Drs. K. Wagschal and C. Lee of the U.S. Department of Agriculture for providing the DeAFc gene, technical support and helpful discussions. We also thank Dr. I. Maiti of the University of Kentucky for providing the Xyln gene and Dr. M. Montross for supplying the corn stover substrates. This work is supported by a grant from the U.S. Department of Agriculture to L.Y. (2006-35504-17413).
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Fan, Z., Werkman, J.R. & Yuan, L. Engineering of a multifunctional hemicellulase. Biotechnol Lett 31, 751–757 (2009). https://doi.org/10.1007/s10529-009-9926-3
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DOI: https://doi.org/10.1007/s10529-009-9926-3