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Characterization and evolution of xylose isomerase screened from the bovine rumen metagenome in Saccharomyces cerevisiae

Paper ID Volume ID Publish Year Pages File Format Full-Text
20014 43153 2016 6 PDF Available
Title
Characterization and evolution of xylose isomerase screened from the bovine rumen metagenome in Saccharomyces cerevisiae
Abstract

•Xylose isomerases genes were screened for efficient expression in Saccharomyces cerevisiae.•The xylose isomerase from bovine rumen exhibited high activity when expressing in S. cerevisiae.•A variant containing two mutations (K11T/D220V) of Ru-XI that exhibited 68% increase in enzyme activity was isolated.

The conversion of abundant levels of xylose in lignocellulosic materials into viable products would generate economic benefits. The heterologous expression of the xylose isomerase (XI) gene is considered a direct and effective strategy for establishing the xylose metabolic pathway in Saccharomyces cerevisiae. However, only limited sources of xylA are functionally expressed in S. cerevisiae and are capable of driving effective xylose consumption. In this study, Ru-xylA (where Ru represents the rumen), which was screened from the contents of the bovine rumen metagenomic library, was functionally expressed in S. cerevisiae, and the enzyme activity was 1.31 U mg−1 protein. This is a new source of XI that can exhibit high activity levels in S. cerevisiae. The activity of this enzyme is comparable to those of the Piromyces sp. XI. Then, the Ru-XI activity was further improved through mutagenesis and growth-based screening in a centromeric plasmid. A variant containing two mutations (K11T/D220V) that exhibited a 68% increase in enzyme activity was isolated. Our work identified a new xylose isomerase that can be functionally expressed in S. cerevisiae and results in a higher XI enzyme activity through mutagenesis.

Keywords
Budding yeast; Xylose isomerase; Bovine rumen metagenome; Growth-based screening; Ethanol
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Characterization and evolution of xylose isomerase screened from the bovine rumen metagenome in Saccharomyces cerevisiae
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 121, Issue 2, February 2016, Pages 160–165
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