Purification, cloning, sequencing and over-expression in Escherichia coli of a regioselective aliphatic nitrilase from Acidovorax facilis 72W

A regioselective aliphatic nitrilase from Acidovorax facilis 72W was purified and characterized, and the corresponding gene was cloned and sequenced. This nitrilase gene was over-expressed in Escherichia coli, generating a microorganism that efficiently and regioselectively catalyzes the conversion...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2003-04, Vol.61 (2), p.118-122
Main Authors: Chauhan, S, Wu, S, Blumerman, S, Fallon, R.D, Gavagan, J.E, DiCosimo, R, Payne, M.S
Format: Article
Language:English
Subjects:
Acidovorax facilis
Acids
Amino Acid Sequence
Aminohydrolases - chemistry
Aminohydrolases - genetics
Aminohydrolases - isolation & purification
Aminohydrolases - metabolism
Base Sequence
Betaproteobacteria - enzymology
Betaproteobacteria - genetics
biocatalysts
Biological and medical sciences
Biotechnology
Chromatography
Cloning
Cloning, Molecular
E coli
Enzymes
Escherichia coli
Escherichia coli - enzymology
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
gene overexpression
genes
Genetics
industrial applications
Methods. Procedures. Technologies
Microbial engineering. Fermentation and microbial culture technology
Microbiological synthesis
Molecular biology
Molecular Sequence Data
nitrilase
Peptides
Physiological aspects
Plasmids
Potassium
Properties
Proteins
Sequence Analysis, DNA
Stereoisomerism
Studies
Substrate Specificity
Transferases
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Summary:A regioselective aliphatic nitrilase from Acidovorax facilis 72W was purified and characterized, and the corresponding gene was cloned and sequenced. This nitrilase gene was over-expressed in Escherichia coli, generating a microorganism that efficiently and regioselectively catalyzes the conversion of aliphatic dinitriles to cyanocarboxylic acids. The high yields obtained, mild reaction conditions used, and robustness observed make this biocatalyst suitable for industrial applications.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-002-1192-4