The importance of porE and porF in the anabolic pyruvate oxidoreductase of Methanococcus maripaludis.
The operon of the anabolic pyruvate oxidoreductase (POR) of Methanococcus maripaludis encodes two genes ( porEF) whose functions are unknown. Because these genes possess sequence similarity to polyferredoxins, they may be electron carriers to the POR. To elucidate whether the methanococcal POR requires PorEF for activity, a deletion mutant, strain JJ150, lacking porEF was constructed. Compared to the wild-type strain JJ1, the mutant grew more slowly in minimal medium and minimal plus acetate medium, and pyruvate-dependent methanogenesis was inhibited. In contrast, the methyl-viologen-dependent pyruvate-oxidation activity of POR, carbon monoxide dehydrogenase, and hydrogenase activities of the mutant were similar to those of the wild-type. Upon genetic complementation of the mutant with porEF in the methanococcal shuttle vector pMEV2+ porEF, growth in minimal medium and pyruvate-dependent methanogenesis were restored to wild-type levels. Complementation with porE alone restored methanogenesis from pyruvate but not growth in minimal medium. Complementation with porF alone partially restored growth but not methanogenesis from pyruvate. Although the specific roles of porE and porF have not been determined, these results suggest that PorEF play important roles in the anabolic POR in vivo even though they are not required for the dye-dependent activity.
Lin
Winston
W
Department of Microbiology, University of Georgia, Athens, GA 30602-2605, USA.
Whitman
William B
WB
D000445:Aldehyde Oxidoreductases / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D023061:Gene Order; D019766:Genes, Archaeal; D005816:Genetic Complementation Test; D006864:Hydrogenase / Q000378:metabolism; D007658:Ketone Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D008697:Methane / Q000378:metabolism; D017017:Methanococcus / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D009097:Multienzyme Complexes / Q000378:metabolism; D009876:Operon; D050743:Pyruvate Synthase; D019289:Pyruvic Acid / Q000378:metabolism
D019843:Archaeal Proteins; D009097:Multienzyme Complexes; D019289:Pyruvic Acid; D006864:Hydrogenase; D000445:Aldehyde Oxidoreductases; D007658:Ketone Oxidoreductases; D050743:Pyruvate Synthase; C019823:carbon monoxide dehydrogenase; D008697:Methane