Elimination of manganese(II, III) oxidation in pseudomonas putida GB-1 by a double knockout of two putative multicopper oxidase genes

Kati Geszvain, James K. McCarthy, Bradley Tebo

Research output: Contribution to journalArticle

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Abstract

Bacterial manganese(II) oxidation impacts the redox cycling of Mn, other elements, and compounds in the environment; therefore, it is important to understand the mechanisms of and enzymes responsible for Mn(II) oxidation. In several Mn(II)-oxidizing organisms, the identified Mn(II) oxidase belongs to either the multicopper oxidase (MCO) or the heme peroxidase family of proteins. However, the identity of the oxidase in Pseudomonas putida GB-1 has long remained unknown. To identify the P. putida GB-1 oxidase, we searched its genome and found several homologues of known or suspected Mn(II) oxidase-encoding genes (mnxG, mofA, moxA, and mopA). To narrow this list, we assumed that the Mn(II) oxidase gene would be conserved among Mn(II)-oxidizing pseudomonads but not in nonoxidizers and performed a genome comparison to 11 Pseudomonas species. We further assumed that the oxidase gene would be regulated by MnxR, a transcription factor required for Mn(II) oxidation. Two loci met all these criteria: PputGB1_2447, which encodes an MCOhomologous to MnxG, and PputGB1_2665, which encodes anMCOwith very low homology to MofA. In-frame deletions of each locus resulted in strains that retained some ability to oxidize Mn(II) or Mn(III); loss of oxidation was attained only upon deletion of both genes. These results suggest that PputGB1_2447 and PputGB1_2665 encode twoMCOsthat are independently capable of oxidizing both Mn(II) and Mn(III). The purpose of this redundancy is unclear; however, differences in oxidation phenotype for the single mutants suggest specialization in function for the two enzymes.

Original languageEnglish (US)
Pages (from-to)357-366
Number of pages10
JournalApplied and Environmental Microbiology
Volume79
Issue number1
DOIs
StatePublished - Jan 2013

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Pseudomonas putida
Manganese
manganese
Oxidoreductases
oxidation
gene
Genes
genes
moxonidine
genome
enzyme
loci
gene deletion
enzymes
Genome
homology
Pseudomonas
phenotype
peroxidase
Gene Deletion

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology

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Elimination of manganese(II, III) oxidation in pseudomonas putida GB-1 by a double knockout of two putative multicopper oxidase genes. / Geszvain, Kati; McCarthy, James K.; Tebo, Bradley.

In: Applied and Environmental Microbiology, Vol. 79, No. 1, 01.2013, p. 357-366.

Research output: Contribution to journalArticle

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