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  • Archaea  (2)
  • 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
    ISSN: 1432-072X
    Keywords: Key words     Thermoproteus tenax ; Pyrobaculum ; islandicum ; Hyperthermophiles ; Archaea ; Acetyl-CoA oxidation ; Citric acid cycle ; Sulfur respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract      The oxidation of organic compounds with elemental sulfur or thiosulfate as electron acceptor was studied in the anaerobic hyperthermophilic archaea Thermoproteus tenax and Pyrobaculum islandicum. T. tenax was grown on either glucose or casamino acids and sulfur; P. islandicum on peptone and either elemental sulfur or thiosulfate as electron acceptor. During exponential growth only CO 2 and H2S rather than acetate, alanine, lactate, and succinate were detected as fermentation products of both organisms; the ratio of CO2/H2S formed was 1 : 2 with elemental sulfur and 1 : 1 with thiosulfate as electron acceptor. Cell extracts of T. tenax and P. islandicum contained all enzymes of the citric acid cycle in catabolic activities: citrate synthase, aconitase, isocitrate dehydrogenase (NADP+ -reducing), oxoglutarate : benzylviologen oxidoreductase, succinyl-CoA synthetase, succinate dehydrogenase, fumarase and malate dehydrogenase (NAD+-reducing). Carbon monoxide dehydrogenase activity was not detected. We conclude that in T. tenax and P. islandicum organic compounds are completely oxidized to CO2 with sulfur or thiosulfate as electron acceptor and that acetyl-CoA oxidation to CO2 proceeds via the citric acid cycle.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 11 (1995), S. 26-57 
    ISSN: 1573-0972
    Keywords: Acetate formation ; acetyl-CoA oxidation ; Archaea ; Bacteria ; chemolithoautotroph ; chemoorganoheterotroph ; glycolytic pathway ; hyperthermophiles ; metabolic pathways ; peptide metabolism ; sugar metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Hyperthermophiles are characterized by a temperature optimum for growth between 80 and 110°C. They are considered to represent the most ancient phenotype of living organisms and thus their metabolic design might reflect the situation at an early stage of evolution. Their modes of metabolism are diverse and include chemolithoautotrophic and chemoorganoheterotrophic. No extant phototrophic hyperthermophiles are known. Lithotrophic energy metabolism is mostly anaerobic or microaerophilic and based on the oxidation of H2 or S coupled to the reduction of S, SO inf4 sup2- , CO2 and NO inf3 sup- but rarely to O2. the substrates are derived from volcanic activities in hyperthermophilic habitats. The lithotrophic energy metabolism of hyperthermophiles appears to be similar to that of mesophiles. Autotrophic CO2 fixation proceeds via the reductive citric acid cycle, considered to be one of the first metabolic cycles, and via the reductive acetyl-CoA/carbon monoxide dehydrogenase pathway. The Calvin cycle has not been found in hyperthermophiles (or any Archaea). Organotrophic metabolism mainly involves peptides and sugars as substrates, which are either oxidized to CO2 by external electron acceptors or fermented to acetate and other products. Sugar catabolism in hyperthermophiles involves non-phosphorylated versions of the Entner-Doudoroff pathway and modified versions of the Embden-Meyerhof pathway. The ‘classical’ Embden-Meyerhof pathway is present in hyperthermophilic Bacteria (Thermotoga) but not in Archaea. All hyperthermophiles (and Archaea) tested so far utilize pyruvate:ferredoxin oxidoreductase for acetyl-CoA formation from pyruvate. Acetyl-CoA oxidation in anaerobic sulphur-reducing and aerobic hyperthermophiles proceeds via the citric acid cycle; in the hyperthermophilic sulphate-reducer Archaeoglobus an oxidative acetyl-CoA/carbon monoxide dehydrogenase pathway is operative. Acetate formation from acetyl-CoA in Archaea, including hyperthermophiles, is catalysed by acetyl-CoA synthetase (ADP-forming), a novel prokarvotic enzyme involved in energy conservation. In Bacteria, including the hyperthermophile Thermotoga, acetyl-CoA conversion to acetate involves two enzymes, phosphate acetyltransferase and acetate kinase.
    Type of Medium: Electronic Resource
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