Identification, functional expression and kinetic analysis of two primary amine oxidases from Rhodococcus opacus

Alexander Foster, Nicole Barnes, Robert Speight, Mark A. Keane

Research output: Contribution to journalArticle

Abstract

Two native copper-containing amine oxidases (EC 1.4.3.21) have been isolated from Rhodococcus opacus and reveal phenotypic plasticity and catalytic activity with respect to structurally diverse natural and synthetic amines. Altering the amine growth substrate has enabled tailored and targeted oxidase upregulation, which with subsequent treatment by precipitation, ion exchange and gel filtration, achieved a 90-150 fold purification. MALDI-TOF mass spectrometric and genomic analysis has indicated multiple gene activation with complex biodegradation pathways and regulatory mechanisms. Additional post-purification characterisation has drawn on the use of carbonyl reagent and chelating agent inhibitors. Michaelis-Menten kinetics for common aliphatic and aromatic amine substrates and several structural analogues demonstrated a broad specificity and high affinity with Michaelis constants (K-M) ranging from 0.1 to 0.9 mM for C-1-C-5 aliphatic mono-amines and <0.2 mM for a range of aromatic amines. Potential exploitation of the enzymatic versatility of the two isolated oxidases in biosensing and bioprocessing is discussed. (C) 2011 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)73-82
Number of pages10
JournalJournal of Molecular Catalysis B: Enzymatic
Volume74
Issue number1-2
DOIs
Publication statusPublished - Jan 2012

Keywords

  • Amine oxidase
  • Rhodococcus opacus
  • Oxidase induction
  • Enzyme purification
  • Oxidative deamination kinetics

Cite this

Foster, Alexander ; Barnes, Nicole ; Speight, Robert ; Keane, Mark A. / Identification, functional expression and kinetic analysis of two primary amine oxidases from Rhodococcus opacus. In: Journal of Molecular Catalysis B: Enzymatic. 2012 ; Vol. 74, No. 1-2. pp. 73-82.
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Identification, functional expression and kinetic analysis of two primary amine oxidases from Rhodococcus opacus. / Foster, Alexander; Barnes, Nicole; Speight, Robert; Keane, Mark A.

In: Journal of Molecular Catalysis B: Enzymatic, Vol. 74, No. 1-2, 01.2012, p. 73-82.

Research output: Contribution to journalArticle

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