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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 144 (1986), S. 78-83 
    ISSN: 1432-072X
    Keywords: Desulfuromonas acetoxidans ; Succinate oxidation ; Sulphur reduction ; Acetate oxidation ; Citric acid cycle ; Reverse of electron transport ; Menaquinone ; ATP synthase ; Succinate dehydrogenase ; Sulphur reductase ; NADH dehydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The oxidation of succinate with elemental sulphur in Desulfuromonas acetoxidans was investigated using a membrane preparation of this bacterium. The following results were obtained: 1. The preparation catalyzed the oxidation of succinate with sulphur and NAD. These reactions were dependent on ATP and were abolished by the presence of protonophores or dicyclohexylcarbodiimide (DCCD). 2. The membrane preparation also catalyzed the reduction of fumarate with H2S or with NADH. These activities were not dependent on ATP and were not affected by protonophores or DCCD. 3. By extraction-reincorporation experiments it could be shown that menaquinone is involved in electron transport between H2S and fumarate and between NADH and fumarate. 4. The membrane fraction catalyzed the reduction of the water-soluble menaquinone-analogue dimethylnaphthoquinone (DMN) by succinate, H2S, or NADH, and the oxidation of DMNH2 by fumarate. These activities were not dependent on the presence of menaquinone and were not influenced by ATP. 5. The activities involving succinate oxidation or fumarate reduction were similarly sensitive to 2(n-nonyl)-4-hydroxyquinoline-N-oxide, while H2S and NADH oxidation by DMN were not affected by the inhibitor. It is concluded that the catabolism of D. acetoxidans involves the energy-driven oxidation of succinate with elemental sulphur or NAD as electron acceptors and that menaquinone is a component of the electron transport chain catalyzing these reactions.
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  • 2
    ISSN: 1432-072X
    Keywords: Desulfobacter postgatei ; Citric acid cycle ; Anaplerotic reactions ; Citrate (si)-synthase ; 2-Oxoglutarate:ferredoxin oxidoreductase ; Succinate dehydrogenase ; Succinyl-CoA:acetate CoA transferase ; Acetyl-CoA synthetase ; Pyruvate synthase ; Phosphoenolpyruvate synthetase ; Phosphoenolpyruvate carboxylase ; Menaquinone ; Ferredoxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The strict anaerobe Desulfobacter postgatei oxidizes acetate to CO2 with sulfate as electron acceptor. During growth at 28°C with a doubling time of 16 h the oxidation and assimilation rate of acetate were 280 nmol and 20 nmol per min and mg protein, respectively. In cell extracts all the enzymes of the citric acid cycle were found (numbers in brackets=specific activities in nmol per min and mg protein at 28°C): Citrate (si)-synthase (250); aconitase (200); NADP-dependent isocitrate dehydrogenase (8500); 2-oxoglutarate: ferredoxin oxidoreductase (300); succinyl-CoA: acetate CoA transferase (160); membrane bound succinate dehydrogenase (3500); and membrane bound malate dehydrogenase with 2,3-dimethyl-1,4-naphthoquinone as artificial electron acceptor (54). The following enzymes catalyzing the synthesis of oxaloacetate from acetate and CO2 were also present: Acetyl-CoA synthetase (10); ferredoxin dependent pyruvate synthase (30); phosphoenolpyruvate synthetase (10); and phosphoenolpyruvate carboxylase (24). The key enzymes of the glyoxylate cycle were not detected. The order of magnitude of the observed enzyme activities was sufficient to account for an oxidation of acetate via the citric acid cycle and for a synthesis of oxaloacetate from acetate and CO2 as anaplerotic reaction. The membranes of D. postgatei contained menaquinone (0.35 nmol per mg cell dry weight) rather than ubiquinone or demethylmenaquinone. The cytoplasmic fraction contained ferredoxin (0.09 nmol per mg cell dry weight).
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-072X
    Keywords: Sulfate-reducing bacteria ; Desulfobacterium ; Desulfotomaculum ; Acetate oxidation ; Citric acid cycle ; Acetyl CoA pathway ; CO dehydrogenase ; Formate dehydrogenase ; Mini-methane formation ; Methane oxidation ; Fluoroacetate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In several sulfate-reducing bacteria capable of complete oxidation of acetate (or acetyl CoA), the citric acid cycle is not operative. No 2-oxoglutarate dehydrogenase activity was found in these organisms, and the labelling pattern of oxaloacetate excludes its synthesis via 2-oxo-glutarate. These sulfate-reducers contained, however, high activities of the enzymes carbon monoxide dehydrogenase and formate dehydrogenase and catalyzed an isotope exchange between CO2 and the carboxyl group of acetate (or acetyl CoA), showing a direct C-C-cleavage of activated acetic acid. These findings suggest that in the investigated sulfate-reducers acetate is oxidized to CO2 via C1 intermediates. The proposed pathway provides a possible explanation for the reported different fluoroacetate sensitivity of acetate oxidation by anaerobic bacteria, for mini-methane formation, as well as for the postulated anaerobic methane oxidation by special sulfate-reducers.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 150 (1988), S. 374-380 
    ISSN: 1432-072X
    Keywords: Desulfotomaculum ; Sulfate reduction ; Acetate oxidation ; Acetyl-CoA ; Carbon monoxide dehydrogenase ; Tetrahydrofolates ; Acetyl-CoA/carbon monoxide dehydrogenase pathway ; Citric acid cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Desulfotomaculum acetoxidans has been proposed to oxidize acetate to CO2 via an oxidative acetyl-CoA/carbon monoxide dehydrogenase pathway rather than via the citric acid cycle. We report here the presence of the enzyme activities required for the operation of the novel pathway. In cell extracts the following activities were found (values in brackets=specific activities and apparent K m; 1 U·mg-1=1 μmol·min-1·mg protein-1 at 37°C): Acetate kinase (6.3 U·mg-1; 2 mM acetate; 2.4 mM ATP); phosphate acetyltransferase (60 U·mg-1, 0.4 mM acetylphosphate; 0.1 mM CoA); carbon monoxide dehydrogenase (29 U·mg-1; 13% carbon monoxide; 1.3 mM methyl viologen); 5,10-methylenetetrahydrofolate reductase (3 U·mg-1, 0.06 mM CH3−FH4); methylenetetrahydrofolate dehydrogenase (3.6 U·mg-1, 0.9 mM NAD, 0.1 mM CH2=FH4); methenyltetrahydrofolate cyclohydrolase (0.3 U·mg-1); formyltetrahydrofolate synthetase (3 U·mg-1, 1.4 mM FH4, 0.4 mM ATP, 13 mM formate); and formate dehydrogenase (10 U·mg-1, 0.4 mM formate, 0.5 mM NAD). The specific activities are sufficient to account for the in vivo acetate oxidation rate of 0.26 U·mg-1.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 136 (1983), S. 230-233 
    ISSN: 1432-072X
    Keywords: Desulfobacter postgatei ; Acetate oxidation ; Sulfate reduction ; Citric acid cycle ; Anaplerotic reactions ; Citrate (si)-synthesis ; 2-Oxoglutarate synthesis ; Oxaloacetate synthesis ; Pyruvate synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract All the enzymes required for the oxidation of acetate to CO2 via the citric acid cycle were found in Desulfobacter postgatei. To obtain in vivo evidence for the operation of this cycle, the sulfate reducing bacterium was grown on [14C]acetate in the presence of a large pool of 12CO2 and the incorporation of 14C into glutamate (≙ 2-oxoglutarate), aspartate (≙ oxaloacetate), and alanine (≙ pyruvate) was studied. The labelling data were found to be consistent with (i) the oxidation of acetate to CO2 via the reactions of the citric acid cycle, (ii) the synthesis of citrate via a citrate (si)-synthase, and (iii) the anaplerotic synthesis of oxaloacetate from acetate and 2 CO2 via pyruvate as intermediate.
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  • 6
    ISSN: 1432-072X
    Keywords: Desulfuromonas acetoxidans ; Acetate oxidation ; Sulfur reduction ; Acetate activation ; Citric acid cycle ; Anaplerotic reaction ; Citrate (si)-synthase ; Succinate dehydrogenase ; Succinyl-CoA: acetate CoA transferase ; 2-Oxoglutarate: ferredoxin oxidoreductase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The strict anaerobe Desulfuromonas acetoxidans can oxidize acetate to CO2 with elemental sulfur as electron acceptor. 14C-labelling experiments and enzyme studies are described revealing that acetate oxidation proceeds via the citric acid cycle with the synthesis of oxaloacetate from acetate and 2 CO2 via pyruvate as anaplerotic reaction. An oxidation of acetate via one carbon unit intermediates as proposed for anaerobic bacteria fermenting acetate to 2 CO2 and 4 H2 was excluded.
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  • 7
    ISSN: 1432-072X
    Keywords: Acetate oxidation ; Citric acid cycle ; ATPase ; Energy conservation ; Electron transport ; Ferredoxin: NADP oxidoreductase ; NADPH dehydrogenase ; Succinate dehydrogenase ; Malate dehydrogenase ; Heptylhydroxyquinoline-N-oxide ; Sulfate-reducing bacteria ; Desulfobacter postgatei
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Desulfobacter postgatei grows on acetate and sulfate as energy source. The oxidation of acetate to 2 CO2 proceeds via the citric acid cycle involving membrane-bound succinate dehydrogenase and membrane-bound malate dehydrogenase. We report here that the organism contains membrane-bound NADPH dehydrogenase and ferredoxin: NADP oxidoreductase for the reoxidation of NADPH and reduced ferredoxin generated during isocitrate- and 2-oxoglutarate oxidation, respectively. The presence of proton translocating ATPase activity is also described. NADPH dehydrogenase and succinate dehydrogenase were found to be electrically connected within the membrane and electron transfer between these two enzymes was shown to be coupled with proton translocation. The membrane fraction catalyzed the oxidation of NADPH with fumarate and the reduction of NADP with succinate. NADPH oxidation with fumarate was stimulated by protonophores and inhibited by the proton translocating ATPase inhibitor dicyclohexylcarbodiimide (DCCD) and by heptylhydroxyquinoline-N-oxide (HQNO); inhibition by DCCD was relieved by protonophores. NADP reduction with succinate was dependent on ATP and inhibited by protonophores, DCCD, and HQNO. The membrane fraction also mediated the oxidation of NADPH with the water soluble menaquinone analogue dimethylnaphthoquinone (DMN) and the reduction of fumarate with DMNH2. Only the former reaction was stimulated by protonophores and only the latter reaction was inhibited by HQNO. This suggests that the NADPH dehydrogenase reaction is the site of energy conservation and the succinate dehydrogenase is the site of HQNO inhibition.
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