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Structural perspectives of an engineered β-1,4-xylanase with enhanced thermostability

Paper ID Volume ID Publish Year Pages File Format Full-Text
23069 43415 2014 8 PDF Available
Title
Structural perspectives of an engineered β-1,4-xylanase with enhanced thermostability
Abstract

•Complex structures of a GH10 XynAS9 and its thermostable mutant were solved.•Mutant XynAS9 forms more intramolecular interactions than the wild type protein.•Mutant XynAS9 is more rigid than the wild type protein.

The glycoside hydrolase 10 (GH10) xylanase from Streptomyces sp. 9 (XynAS9) can operate in a broad range of pH and temperature, and thus is a potential candidate for commercial applications. Recently, we engineered XynAS9 via mutating several residues in accordance with the consensus sequences of GH10 thermophilic xylanases in an attempt to improve the enzyme thermostability and thermotolerance. The most promising effects were observed in the double mutant V81P/G82E. In order to investigate the molecular mechanism of the improved thermal profile of XynAS9, complex crystal structures of the wild type (WT) and mutant (MT) enzyme were solved at 1.88–2.05 Å resolution. The structures reveal a classical GH10 (β/α)8 TIM-barrel fold. In MT XynAS9, E82 forms several interactions to its neighboring residues, which might aid in stabilizing the local structure. Furthermore, the MT structure showed lower B factors for individual residues compared to the WT structure, reflecting the increased MT protein rigidity. Analyses of the XynAS9 structures also delineate the detailed enzyme–substrate interaction network. More importantly, possible explanations for the enhanced thermal profiles of MT XynAS9 are proposed, which may be a useful strategy for enzyme engineering in the future.

Keywords
β-1,4-Xylanase; Crystal structure; Thermozyme; Protein engineering; Protein rigidity
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Structural perspectives of an engineered β-1,4-xylanase with enhanced thermostability
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 189, 10 November 2014, Pages 175–182
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us