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  • ATP formation  (1)
  • Key words:Methanosarcina barkeri– Pyruvate-utilizing mutant – Methanogenesis – Archaea – Pyruvate fermentation – Acetate fermentation – Growth yields (YCH4) – Ferredoxin – Pyruvate: ferredoxin oxidoreductase  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 161 (1994), S. 33-46 
    ISSN: 1432-072X
    Keywords: Key words:Methanosarcina barkeri– Pyruvate-utilizing mutant – Methanogenesis – Archaea – Pyruvate fermentation – Acetate fermentation – Growth yields (YCH4) – Ferredoxin – Pyruvate: ferredoxin oxidoreductase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Methanosarcina barkeri strain Fusaro was found to grow on pyruvate as sole carbon and energy source after an incubation period of 10 – 12 weeks in the presence of high pyruvate concentrations (100 mM). Growth studies, cell suspension experiments and enzymatic investigations were performed with pyruvate-utilizing M. barkeri. For comparison acetate-adapted cells of M. barkeri were analyzed. 1. Pyruvate-utilizing M. barkeri grew on pyruvate (100 mM) with an initial doubling time of about 25 h (37 °C, pH 6.5) up to cell densities of about 0.8 g cell dry weight/l. The specific growth rate was linearily dependent on the pyruvate concentration up to 100 mM indicating that pyruvate was taken up by passive diffusion. Only CO2 and CH4 were detected as fermentation products. As calculated from fermentation balances pyruvate was converted to CH4 and CO2 according to following equation: Pyruvate−+H++0.5 H2O→1.25 CH4+1.75 CO2. The molar growth yield (YCH4) was about 14 g dry weight cells/mol CH4. In contrast the growth yield (YCH4) of M. barkeri during growth on acetate (Acetate−+H+→CH4+CO2) was about 3 g/mol CH4. 2. Cell suspensions of pyruvate-grown M. barkeri catalyzed the conversion of pyruvate to CH4, CO2 and H2 (5 – 15 nmol pyruvate consumed/min×mg protein). At low cell concentrations (0.5 mg protein/ml) 1 mol pyruvate was converted to 1 mol CH4, 2 mol CO2 and 1 mol H2. At higher cell concentration less H2 and CO2 and more CH4 were formed due to CH4 formation from H2/CO2. The rate of pyruvate conversion was linearily dependent on the pyruvate concentration up to about 30 mM. Cell suspensions of acetate-grown M. barkeri also catalyzed the conversion of 1 mol pyruvate to 1 mol CH4, 2 mol CO2 and 1 mol H2 at similar rates and with similar affinity for pyruvate as pyruvate-grown cells. 3. Cell extracts of both pyruvate-grown and acetate-grown M. barkeri contained pyruvate: ferredoxin oxidoreductase. The specific activity in pyruvate-grown cells (0.8 U/mg) was 8-fold higher than in acetate-grown cells (0.1 U/mg). Coenzyme F420 was excluded as primary electron acceptor of pyruvate oxidoreductase. Cell extracts of pyruvate-grown M. barkeri contained carbon monoxide dehydrogenase activity and hydrogenase activity catalyzing the reduction by carbon monoxide and hydrogen of both methylviologen and ferredoxin (from Clostridium). This is the first report on growth of a methanogen on pyruvate as sole carbon and energy source, i.e. on a substrate more complex than acetate.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 150 (1988), S. 447-451 
    ISSN: 1432-072X
    Keywords: Acetobacterium woodii ; Caffeate reduction ; ATP formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have addressed the question, whether the reduction of caffeate in Acetobacterium woodii strain NZva16 is coupled to ATP synthesis by electron transport phosphorylation. The following results were obtained: 1. Cultures of A. woodii with H2 and CO2, grew to greater cell densities, when caffeate was also present. Caffeate was reduced to give hydrocaffeate and less acetate was formed. The cell yield based on the amount of caffeate reduced was approximately 1 g dry cells/mol. 2. Non-growing bacterial suspensions catalyzed the reduction of caffeate by H2. The specific activity (0.2–1.0 μmol · min−1 · mg−1 bacterial protein) was as high as expected for a catabolic reaction. 3. The ATP content of bacteria incubated, with H2 increased from 〈 1 to about 7 μmol per g cellular protein on the addition of caffeate. The ATP yield was calculated as 0.06 mol ATP · mol−1 caffeate from the initial velocity of ATP formation and the activity of caffeate reduction. Valinomycin together with nigericin inhibited ATP formation and caused a 2–3-fold increase of the activity of caffeate reduction. Protonophores were without, effect. 4. Caffeate in the presence of H2 caused the uptake of tetraphenylphosphonium cation by the bacteria. The uptake was abolished by valinomycin plus nigericin, and was considerably enhanced by monensin. Protonophores were without effect, even in the presence of monensin. It is concluded that caffeate reduction by H2 is coupled to ATP formation by electron transport phosphorylation. However, the failure of protonophores to prevent phosphorylation and TPP uptake cannot be explained.
    Type of Medium: Electronic Resource
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