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Isolation and mutation of recombinant EPSP synthase from pathogenic bacteria Pseudomonas aeruginosa

Paper ID Volume ID Publish Year Pages File Format Full-Text
36013 45118 2007 7 PDF Available
Title
Isolation and mutation of recombinant EPSP synthase from pathogenic bacteria Pseudomonas aeruginosa
Abstract

The Pseudomonas aeruginosa aroA gene encodes an enzyme called 5-enol-pyruvylshikimate-3-phosphate (EPSP) synthase, which has been shown as the primary target of the herbicide glyphosate. We have cloned this gene and constructed a system for the high level expression of a recombinant form of this enzyme by amplifying the aroA gene from the P. aeruginosa genomic DNA and subcloning into a vector suitable for expression in Escherichia coli. The resulting plasmid, pTrcPA, produced the EPSP synthase in large quantities which has been purified to homogeneity. Furthermore, the site-directed mutants of P. aeruginosa ESPS synthase have been constructed in order to compare in vitro glyphosate sensitivity between the wild-type and the mutant enzymes. The kcat and Km values for substrates in both forward and reverse reactions were obtained from both wild-type and mutant EPSP synthases.

Keywords
EPSP synthase; P. aeruginosa; Isolation; Mutation
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Isolation and mutation of recombinant EPSP synthase from pathogenic bacteria Pseudomonas aeruginosa
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 42, Issue 4, April 2007, Pages 592–598
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