Improving the thermostability of feruloyl esterase by DNA shuffling and site-directed mutagenesis. Issue 11 (November 2015)

• Record Type: Journal Article
• Authors: Li, Jing-Jing; Pei, Xiao-Qiong; Zhang, Shuai-Bing; Wu, Zhong-Liu
• Is Part Of: Process biochemistry. Volume 50:Issue 11(2015:Nov.)
• Page Start: 1783
• Page End: 1787
• Publication Date: 2015-11
• Subjects: Feruloyl esterase -- Thermostability -- DNA shuffling -- Pichia pastoris; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2015.08.009
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: A DNA shuffling library was constructed from four feruloyl esterases. Chimeras with enhanced thermal stability were identified. Half-life of the best chimera increased to 22-fold of the most stable parental FAE. Release of ferulic acid increases to 3.6–12.8-fold using the best chimera. Abstract: 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 °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.