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Engineering a high-performance, metagenomic-derived novel xylanase with improved soluble protein yield and thermostability

Paper ID Volume ID Publish Year Pages File Format Full-Text
17047 42636 2015 7 PDF Available
Title
Engineering a high-performance, metagenomic-derived novel xylanase with improved soluble protein yield and thermostability
Abstract

•A novel metagenomic-derived xylanase gene xyl7 was isolated and expressed.•The enzyme XYL7 showed higher specific activity and broad pH activity range.•A truncated XYL7-Tr3 with tenfold increase in soluble protein yield was acquired.•Mutants showed 250-fold and 10 °C increase in half-life and optimal temperature.•The reducing sugar produced from xylan by mutant XYL7-TC was increased fourfold.

The novel termite gut metagenomic-derived GH11 xylanase gene xyl7 was expressed in Escherichia coli BL21, and the purified XYL7 enzyme exhibited high specific activity (6340 U/mg) and broad pH active range of 5.5–10.0. Directed evolution was employed to enhance the thermostability of XYL7; two mutants (XYL7-TC and XYL7-TS) showed a 250-fold increase in half-life at 55 °C, with a 10 °C increase in optimal temperature compared to that of wild-type XYL7. A truncated enzyme (XYL7-Tr3) acquired by protein engineering showed similar catalytic properties as the wild-type, with a tenfold increase in soluble protein yield by the mutant. The reducing sugar produced by XYL7-TC was about fourfold greater than that produced by their parents when incubated with xylan at 60 °C for 4 h. The engineered novel xylanase exhibited superior enzymatic performance and showed promise as an excellent candidate for industrial application due to its high specific activity, stability and soluble protein yield.

Keywords
Directed molecular evolution; GH11 xylanase; Metagenomic; Protein engineering; Thermostability
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Engineering a high-performance, metagenomic-derived novel xylanase with improved soluble protein yield and thermostability
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 70, March 2015, Pages 35–41
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