Journal
PROCESS BIOCHEMISTRY
Volume 50, Issue 11, Pages 1783-1787Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2015.08.009
Keywords
Feruloyl esterase; Thermostability; DNA shuffling; Pichia pastoris
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Funding
- National Natural Science Foundation of China [21372216, 21572220]
- Open Fund of Key Laboratory of Environmental and Applied Microbiology [KLCAS-2013-06, KLCAS-2014-05, KLCAS-2015-01]
- Key Research Program of the Chinese Academy of Sciences [KGZD-EW-606-14]
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Feruloyl esterases (FAE) with high thermostability are greatly desired for efficient and economical industrial processes. A DNA shuffling strategy was applied to generate a chimeric library from four homologous FAEs originated from fungi. Chimeras with enhanced thermal stability compared to the parental FAEs were identified, which contained segments from 3 or 4 of the parental enzymes. After further incorporation of 2-4 additional beneficial mutations, the half-lives of inactivation of the chimeras at 65 degrees C were increased by up to 22 fold when compared to the most stable parental FAE. As a result, the release of ferulic acid from steam-exploded corn stalk using the best chimera was enhanced by 12.8-fold when compared to the best parental FAE. (C) 2015 Elsevier Ltd. All rights reserved.
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