PMID: 9171390Jun 1, 1997Paper

Purification, characterization, and properties of an aryl aldehyde oxidoreductase from Nocardia sp. strain NRRL 5646

Journal of Bacteriology
T Li, J P Rosazza

Abstract

An aryl aldehyde oxidoreductase from Nocardia sp. strain NRRL 5646 was purified 196-fold by a combination of Mono-Q, Reactive Green 19 agarose affinity, and hydroxyapatite chromatographies. The purified enzyme runs as a single band of 140 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular mass was estimated to be 163 +/- 3.8 kDa by gel filtration, indicating that this enzyme is a monomeric protein. The binding of the enzyme to Reactive Green 19 agarose was Mg2+ dependent. The binding capacity was estimated to be about 0.2 mg of Reactive Green agarose per ml in the presence of 10 mM MgCl2. This enzyme can catalyze the reduction of a wide range of aryl carboxylic acids, including substituted benzoic acids, phenyl-substituted aliphatic acids, heterocyclic carboxylic acids, and polyaromatic ring carboxylic acids, to produce the corresponding aldehydes. The Km values for benzoate, ATP, and NADPH were determined to be 645 +/- 75, 29.3 +/- 3.1, and 57.3 +/- 12.5 microM, respectively. The Vmax was determined to be 0.902 +/- 0.04 micromol/min/mg of protein. Km values for (S)-(+)-alpha-methyl-4-(2-methylpropyl)-benzeneacetic acid (ibuprofen) and its (R)-(-) isomer were determined to be 155 +/- 18 and 34.5 +/-...Continue Reading

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Citations

Nov 15, 2006·The Journal of Biological Chemistry·Padmesh VenkitasubramanianJohn P N Rosazza
Apr 17, 2014·Biotechnology Journal·Kamila Napora-WijataMargit Winkler
Oct 20, 2015·Biotechnology and Bioengineering·Matthew MouraKeith E J Tyo
Mar 9, 2018·Frontiers in Microbiology·Holly StolterfohtMargit Winkler
Jan 13, 2015·Applied and Environmental Microbiology·Aditya M Kunjapur, Kristala L J Prather
Jul 23, 2009·International Journal of Systematic and Evolutionary Microbiology·Andrew S LammJohn P N Rosazza
Nov 10, 2012·Chemical Communications : Chem Comm·Yan NiFrank Hollmann
Aug 2, 2017·Biotechnology Journal·Anna N KhusnutdinovaAlexander F Yakunin
Aug 9, 2006·Critical Reviews in Microbiology·Sindhu Mathew, T Emilia Abraham
Feb 2, 2000·Applied and Environmental Microbiology·T Li, J P Rosazza
Oct 20, 2019·Journal of Biotechnology·Neil Butler, Aditya M Kunjapur
Nov 12, 2017·Current Opinion in Chemical Biology·Margit Winkler

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