Product isotope effects on in vivo methanogenesis by Methanobacterium thermoautotrophicum.
The hydrogen in methane produced by cultures of Methanobacterium thermoautotrophicum originates from water. In H2O/D2O mixtures, a methane product isotope effect is observed that increases rapidly as the water deuterium enrichment approaches 100%. This effect is due to the intracellular production of protons from H2, catalyzed by hydrogenase, which occurs at 12% the rate of water diffusion through the cell membrane. We estimate that water diffusion through the thick cell membrane of M. thermoautotrophicum is retarded by a factor of 10(6) over the free diffusion rate. The intracellular production of H+ suggests that either (1) hydrogenase is not directly involved in the production of a chemiosmotic proton gradient or (2) if it is involved, the proton gradient exists between the cytosol and the interior of vesicles observed in this bacterium. The intrinsic deutrium product isotope effect in methanogenesis is 1.20 +/- 0.1, comparable to anabolic deuterium product isotope effects in other autotrophs. An algebraic model incorporating the intracellular H2 to H+ flux accurately predicts the distribution of deuterated methane species at all levels of water deuterium enrichment.
D003903:Deuterium; D004912:Erythrocytes / Q000378:metabolism; D019605:Euryarchaeota / Q000378:metabolism*; D006864:Hydrogenase; D007553:Isotope Labeling; D008081:Liposomes; D008433:Mathematics; D008697:Methane / Q000378:metabolism*; D008954:Models, Biological; D010088:Oxidoreductases / Q000378:metabolism; D010713:Phosphatidylcholines
D008081:Liposomes; D010713:Phosphatidylcholines; D003903:Deuterium; D010088:Oxidoreductases; D006864:Hydrogenase; D008697:Methane