期刊
BIORESOURCE TECHNOLOGY
卷 117, 期 -, 页码 140-147出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2012.04.042
关键词
Feruloyl esterase; Aspergillus niger; Thermostability; Pichia pastoris; Directed evolution
资金
- Program of 100 Distinguished Young Scientists
- Knowledge Innovation Program [KSCX1-YW-11B2]
- Key Laboratory of Environmental and Applied Microbiology of the Chinese Academy of Sciences [Y1C5101106]
- Province Science Foundation of Sichuan, China [2010SZ0128]
Feruloyl esterase A from Aspergillus niger (AnFaeA) is one of the most important feruloyl esterases of industrial relevance. Previous work aided by the PoPMuSiC algorithm has identified two beneficial mutants (D93G and 5187F) with thermostabilization effect. In this work, twelve additional amino acid substitutions were identified to be beneficial to the thermostability of AnFaeA after screening a random mutagenesis library constructed in Pichia pastoris. Combination of these mutations resulted in a mutant with 80% residual activity after heat treatment at 90 degrees C for 15 min and a half-life increasing from 15 min to >4000 min at 55 degrees C. The thermostabilized mutant displayed significantly enhanced performance compared to the parental AnFaeA when applied to the treatment of steam-exploded corn stalk at 60 degrees C together with an xylanase, demonstrating its great potential for industrial application. (C) 2012 Elsevier Ltd. All rights reserved.
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