Identification and catalytic residues of the arsenite methyltransferase from a sulfate-reducing bacterium, Clostridium sp. BXM

FEMS Microbiol Lett. 2015 Jan;362(1):1-8. doi: 10.1093/femsle/fnu003. Epub 2014 Dec 4.

Abstract

Arsenic methylation is an important process frequently occurring in anaerobic environments. Anaerobic microorganisms have been implicated as the major contributors for As methylation. However, very little information is available regarding the enzymatic mechanism of As methylation by anaerobes. In this study, one novel sulfate-reducing bacterium isolate, Clostridium sp. BXM, which was isolated from a paddy soil in our laboratory, was demonstrated to have the ability of methylating As. One putative arsenite S-Adenosyl-Methionine methyltransferase (ArsM) gene, CsarsM was cloned from Clostridium sp. BXM. Heterologous expression of CsarsM conferred As resistance and the ability of methylating As to an As-sensitive strain of Escherichia coli. Purified methyltransferase CsArsM catalyzed the formation of methylated products from arsenite, further confirming its function of As methylation. Site-directed mutagenesis studies demonstrated that three conserved cysteine residues at positions 65, 153 and 203 in CsArsM are necessary for arsenite methylation, but only Cysteine 153 and Cysteine 203 are required for the methylation of monomethylarsenic to dimethylarsenic. These results provided the characterization of arsenic methyltransferase from anaerobic sulfate-reducing bacterium. Given that sulfate-reducing bacteria are ubiquitous in various wetlands including paddy soils, enzymatic methylation mediated by these anaerobes is proposed to contribute to the arsenic biogeochemical cycling.

Keywords: arsenic; conserved cysteine; methylation; methyltransferase; sulfate-reducing bacterium.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arsenic / metabolism*
  • Arsenic / toxicity
  • Catalytic Domain*
  • Cloning, Molecular
  • Clostridium / enzymology*
  • Clostridium / isolation & purification
  • DNA Mutational Analysis
  • Drug Resistance, Bacterial
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Gene Expression
  • Methyltransferases / genetics
  • Methyltransferases / isolation & purification
  • Methyltransferases / metabolism*
  • Mutagenesis, Site-Directed
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Soil Microbiology

Substances

  • Recombinant Proteins
  • Methyltransferases
  • arsenite methyltransferase
  • Arsenic