title: Electron bifurcation involved in the energy metabolism of the acetogenic bacterium Moorella thermoacetica growing on glucose or H2 plus CO2.
abstract: Moorella thermoacetica ferments glucose to three acetic acids. In the oxidative part of the fermentation, the hexose is converted to 2 acetic acids and 2 CO(2) molecules with the formation of 2 NADH and 2 reduced ferredoxin (Fd(red)(2-)) molecules. In the reductive part, 2 CO(2) molecules are reduced to acetic acid, consuming the 8 reducing equivalents generated in the oxidative part. An open question is how the two parts are electronically connected, since two of the four oxidoreductases involved in acetogenesis from CO(2) are NADP specific rather than NAD specific. We report here that the 2 NADPH molecules required for CO(2) reduction to acetic acid are generated by the reduction of 2 NADP(+) molecules with 1 NADH and 1 Fd(red)(2-) catalyzed by the electron-bifurcating NADH-dependent reduced ferredoxin:NADP(+) oxidoreductase (NfnAB). The cytoplasmic iron-sulfur flavoprotein was heterologously produced in Escherichia coli, purified, and characterized. The purified enzyme was composed of 30-kDa (NfnA) and 50-kDa (NfnB) subunits in a 1-to-1 stoichiometry. NfnA harbors a [2Fe2S] cluster and flavin adenine dinucleotide (FAD), and NfnB harbors two [4Fe4S] clusters and FAD. M. thermoacetica contains a second electron-bifurcating enzyme. Cell extracts catalyzed the coupled reduction of NAD(+) and Fd with 2 H(2) molecules. The specific activity of this cytoplasmic enzyme was 3-fold higher in H(2)-CO(2)-grown cells than in glucose-grown cells. The function of this electron-bifurcating hydrogenase is not yet clear, since H(2)-CO(2)-grown cells additionally contain high specific activities of an NADP(+)-dependent hydrogenase that catalyzes the reduction of NADP(+) with H(2). This activity is hardly detectable in glucose-grown cells.
authors.affiliation: Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Huang
authors.forename: Shuning
authors.initials: S
authors.lastname: Wang
authors.forename: Johanna
authors.initials: J
authors.lastname: Moll
authors.forename: Rudolf K
authors.initials: RK
authors.lastname: Thauer
mesh_terms: D002245:Carbon Dioxide / Q000378:metabolism*; D004583:Electrons; D004734:Energy Metabolism / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000378:metabolism*; D006859:Hydrogen / Q000378:metabolism*; D008969:Molecular Sequence Data; D058845:Moorella / Q000378:metabolism*; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism
chemical_list: D002245:Carbon Dioxide; D006859:Hydrogen; D010088:Oxidoreductases; D005947:Glucose