The catalytic mechanism for aerobic formation of methane by bacteria

Nature. 2013 May 2;497(7447):132-6. doi: 10.1038/nature12061. Epub 2013 Apr 24.

Abstract

Methane is a potent greenhouse gas that is produced in significant quantities by aerobic marine organisms. These bacteria apparently catalyse the formation of methane through the cleavage of the highly unreactive carbon-phosphorus bond in methyl phosphonate (MPn), but the biological or terrestrial source of this compound is unclear. However, the ocean-dwelling bacterium Nitrosopumilus maritimus catalyses the biosynthesis of MPn from 2-hydroxyethyl phosphonate and the bacterial C-P lyase complex is known to convert MPn to methane. In addition to MPn, the bacterial C-P lyase complex catalyses C-P bond cleavage of many alkyl phosphonates when the environmental concentration of phosphate is low. PhnJ from the C-P lyase complex catalyses an unprecedented C-P bond cleavage reaction of ribose-1-phosphonate-5-phosphate to methane and ribose-1,2-cyclic-phosphate-5-phosphate. This reaction requires a redox-active [4Fe-4S]-cluster and S-adenosyl-L-methionine, which is reductively cleaved to L-methionine and 5'-deoxyadenosine. Here we show that PhnJ is a novel radical S-adenosyl-L-methionine enzyme that catalyses C-P bond cleavage through the initial formation of a 5'-deoxyadenosyl radical and two protein-based radicals localized at Gly 32 and Cys 272. During this transformation, the pro-R hydrogen from Gly 32 is transferred to the 5'-deoxyadenosyl radical to form 5'-deoxyadenosine and the pro-S hydrogen is transferred to the radical intermediate that ultimately generates methane. A comprehensive reaction mechanism is proposed for cleavage of the C-P bond by the C-P lyase complex that uses a covalent thiophosphate intermediate for methane and phosphate formation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aerobiosis
  • Archaea / metabolism
  • Bacteria / metabolism*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism
  • Biocatalysis*
  • Deoxyadenosines / chemistry
  • Deoxyadenosines / metabolism
  • Electron Spin Resonance Spectroscopy
  • Glycine / chemistry
  • Glycine / metabolism
  • Hydrogen / metabolism
  • Lyases / chemistry
  • Lyases / metabolism
  • Mass Spectrometry
  • Methane / biosynthesis*
  • Methane / chemistry
  • Methane / metabolism
  • Methionine / metabolism
  • Mutant Proteins / chemistry
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism
  • Pentosephosphates / chemistry
  • Pentosephosphates / metabolism
  • S-Adenosylmethionine / metabolism

Substances

  • Bacterial Proteins
  • Deoxyadenosines
  • Mutant Proteins
  • Pentosephosphates
  • S-Adenosylmethionine
  • Hydrogen
  • Methionine
  • Lyases
  • Methane
  • Glycine