Identification of proteins involved in formaldehyde metabolism by Rhodobacter sphaeroides.
Formaldehyde is an intermediate formed during the metabolism of methanol or other methylated compounds. Many Gram-negative bacteria generate formaldehyde from methanol via a periplasmic pyrroloquinoline quinone (PQQ)-dependent dehydrogenase in which the alpha subunit of an alpha(2)beta(2) tetramer has catalytic activity. The genome of the facultative formaldehyde-oxidizing bacterium Rhodobacter sphaeroides encodes XoxF, a homologue of the catalytic subunit of a proposed PQQ-containing dehydrogenase of Paracoccus denitrificans. R. sphaeroides xoxF is part of a gene cluster that encodes periplasmic c-type cytochromes, including CycI, isocytochrome c(2) and CycB (a cyt c(553i) homologue), as well as adhI, a glutathione-dependent formaldehyde dehydrogenase (GSH-FDH), and gfa, a homologue of a glutathione-formaldehyde activating enzyme (Gfa). To test the roles of XoxF, CycB and Gfa in formaldehyde metabolism by R. sphaeroides, we monitored photosynthetic growth with methanol as a source of formaldehyde and whole-cell methanol-dependent oxygen uptake. Our data show that R. sphaeroides cells lacking XoxF or CycB do not exhibit methanol-dependent oxygen uptake and lack the capacity to utilize methanol as a sole photosynthetic carbon source. These results suggest that both proteins are required for formaldehyde metabolism. R. sphaeroides Gfa is not essential to activate formaldehyde, as cells lacking gfa are capable of both methanol-dependent oxygen uptake and growth with methanol as a photosynthetic carbon source.
Wilson
Shondelle M
SM
Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.
Gleisten
Marshall P
MP
Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.
Donohue
Timothy J
TJ
Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002237:Carbohydrate Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D005557:Formaldehyde / Q000378:metabolism*; D017353:Gene Deletion; D000432:Methanol / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010788:Photosynthesis / Q000502:physiology; D010802:Phylogeny; D012242:Rhodobacter sphaeroides / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D005557:Formaldehyde; D002237:Carbohydrate Dehydrogenases; C082601:aldose dehydrogenase (pyrroloquinoline-quinone); D010100:Oxygen; D000432:Methanol