期刊
EUROPEAN JOURNAL OF BIOCHEMISTRY
卷 270, 期 1, 页码 66-75出版社
BLACKWELL PUBLISHING LTD
DOI: 10.1046/j.1432-1033.2003.03362.x
关键词
energy conservation; methanogenesis; proton motive force; sodium motive force; Methanothermobacter thermautotrophicus
The synthesis of formyl-methanofuran and the reduction of the heterodisulfide (CoM-S-S-CoB) of coenzyme M (HS-CoM) and coenzyme B (HS-CoB) are two crucial, H-2-dependent reactions in the energy metabolism of methanogenic archaea. The bioenergetics of the reactions in vivo were studied in chemostat cultures and in cell suspensions of Methanothermobacter thermautotrophicus metabolizing at defined dissolved hydrogen partial pressures (p(H2)). Formyl-methanofuran synthesis is an endergonic reaction (DeltaGdegrees' = +16 kJ.mol(-1)). By analyzing the concentration ratios between formyl-methanofuran and methanofuran in the cells, free energy changes under experimental conditions (DeltaG') were found to range between +10 and +35 kJ.mol(-1) depending on the p(H2) applied. The comparison with the sodium motive force indicated that the reaction should be driven by the import of a variable number of two to four sodium ions. Heterodisulfide reduction (DeltaGdegrees' = -40 kJ.mol(-1)) was associated with free energy changes as high as -55 to -80 kJ.mol(-1) . The values were determined by analyzing the concentrations of CoM-S-S-CoB, HS-CoM and HS-CoB in methane-forming cells operating under a variety of hydrogen partial pressures. Free energy changes were in equilibrium with the proton motive force to the extent that three to four protons could be translocated out of the cells per reaction. Remarkably, an apparent proton translocation stoichiometry of three held for cells that had been grown at p(H2) < 0.12 bar, whilst the number was four for cells grown above that concentration. The shift occurred within a narrow p(H2) span around 0.12 bar. The findings suggest that the methanogens regulate the bioenergetic machinery involved in CoM-S-S-CoB reduction and proton pumping in response to the environmental hydrogen concentrations.
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