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Published on: April 22, 2016
Cloning of Bacillus licheniformis xylanase gene and characterization of recombinant enzyme
Charles C Lee1, Rena E Kibblewhite-Accinelli, Michael R Smith
1USDA-ARS-WRRC, 800 Buchanan St., Albany, CA 94710, USA. Charles.C.Lee@ars.usda.gov
Abstract:
Hemicellulose is a major component of lignocellulose biomass. Complete degradation of this substrate requires several different enzymatic activities, including xylanase. We isolated a strain of Bacillus licheniformis from a hot springs environment that exhibited xylanase activity. A gene encoding a 23-kDa xylanase enzyme, Xyn11, was cloned, and the recombinant protein was expressed in an Escherichia coli host and biochemically characterized. The optimum activity of the enzyme was at pH 5-7 and 40-50 degrees C. The enzyme was stable at temperatures up to 50 degrees C. Against birchwood xylan, the enzyme had an apparent K(m) of 6.7 mg/mL and V(max) of 379 micromol/min/mg.
