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  • 1
    ISSN: 1574-6968
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie
    Notizen: Abstract Respiratory nitrate reductase from the denitrifying bacterium Pseudomonas stutzeri is an iron-sulfur enzyme containing the molybdenum cofactor. Hydrolysis of native nitrate reductase with aqueous sulfuric acid revealed 0.92 mol of 5′-GMP per mol of enzyme. The pterin present in the molybdenum cofactor was liberated from the protein and reacted with iodoacetamide. The resulting di(carboxamidomethyl) (cam) derivative was purified on a C18-cartridge and analyzed for its structural elements. Treatment of the cam derivative with nucleotide pyrophosphatase and subsequent HPLC analysis revealed the formation of di(cam)molybdopterin and 5′-GMP at a 1:1 molar ratio and with a yield of 79% with respect to the molybdenum content of the enzyme. Treatment of the cam derivative with nucleotide pyrophosphatase and alkaline phosphatase led to the liberation of 0.51 mol dephosphodi(cam)molybdopterin and of 0.59 mol guanosine per mol of enzyme, which is equal to a molar ratio of 1:2.2. The results indicate, that the organic moiety of the molybdenum cofactor of nitrate reductase from P. stutzeri is molybdopterin guanine dinucleotide of which one mol is contained per mol of nitrate reductase.
    Materialart: Digitale Medien
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  • 2
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 87 (1990), S. 0 
    ISSN: 1574-6968
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie
    Notizen: Abstract The list of carboxydotrophic bacteria is constantly growing and we have found that burning charcoal piles harbor an especially rich CO-oxidizing microflora. The newly isolated Streptomyces thermoautotrophicus UBT1 is particularly interesting as it is thermophilic, capable of chemolithoautotrophic growth with CO or H2 plus CO2 and incapable of using organic substrates. Molybdenum is essential for CO-autotrophic growth.Some species of carboxydotrophic bacteria can denitrify under heterotrophic conditions yielding N2 (e.g. Pseudomonas carboxydoflava) or N2O (e.g. Pseudomonas carboxydohydrogena); others perform nitrate respiration (e.g. Azomonas B1). P. carboxydohydrogena could grow at the expense of H2 plus CO2 using nitrate as electron acceptor.In intact cells of Pseudomonas carboxydovorans, CO dehydrogenase has the ability of dissociating from and rebinding to the cytoplasmic membrane. That process can be simulated in vitro by removing CO dehydrogenase from cytoplasmic membranes and rebinding it to depleted membranes. Reconstitution of the enzyme onto depleted membranes requiring di- or trivalent cations, was specific for membranes from CO-grown bacteria and led to reactivation of respiratory activities with CO.A complex consisting of 1 molecule of CO dehydrogenase and 2 molecules of cytochrome b561 could be isolated from cytoplasmic membranes of P. carboxydovorans solubilized with dodecyl β-d-maltoside. Within the complex as well as in assays containing purified CO dehydrogenase and cytochrome b561 the latter could serve as an electron acceptor. CO dehydrogenase had hydrogenase activity, and its KM of only 5 mM H2 suggested a role in the formation of H2. P. carboxydovorans OM5 contains the 128-kilo-base pairs (kb) plasmid pHCG3 which is essential for CO- and H2-lithoautotrophic growth. Evidence for the existence of pHCG3-coded structural genes of CO dehydrogenase was obtained from dot blot hybridizations employing synthetic oligodeoxynucleotides as heterologous probes for the detection of the S- and M-subunit genes. Employing appropriate probe genes encoding membrane-bound hydrogenase, ribulose biphosphate carboxylase and phosphoribulokinase were also identified on plasmid pHCG3.
    Materialart: Digitale Medien
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Biodegradation 3 (1992), S. 61-82 
    ISSN: 1572-9729
    Schlagwort(e): carbon monoxide (CO) ; ecology ; enzymes ; groups of CO-utilizing bacteria ; molecular genetics ; pathways
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Energietechnik , Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract The utilization of carbon monoxide as energy and/or carbon source by different physiological groups of bacteria is described and compared. Utilitarian CO oxidation which is coupled to the generation of energy for growth is achieved by aerobic and anaerobic eu- and archaebacteria. They belong to the physiological groups of aerobic carboxidotrophic, facultatively anaerobic phototrophic, and anaerobic acetogenic, methanogenic or sulfate-reducing bacteria. The key enzyme in CO oxidation is CO dehydrogenase which is a molybdo iron-sulfur flavoprotein in aerobic CO-oxidizing bacteria and a nickel-containing iron-sulfur protein in anaerobic ones. In carboxidotrophic and phototrophic bacteria, the CO-born CO2 is fixed by ribulose bisphosphate carboxylase in the reductive pentose phosphate cycle. In acetogenic, methanogenic, and probably in sulfate-reducing bacteria, CODH/acetyl-CoA synthase directly incorporates CO into acetyl-CoA. In plasmid-harbouring carboxidotrophic bacteria, CO dehydrogenase as well as enzymes involved in CO2 fixation or hydrogen utilization are plasmid-encoded. Structural genes encoding CO dehydrogenase were cloned from carboxidotrophic, acetogenic and methanogenic bacteria. Although they are clustered in each case, they are genetically distinct. Soil is a most important biological sink for CO in nature. While the physiological microbial groups capable of CO oxidation are well known, the type and nature of the microorganisms actually representing this sink are still enigmatic. We also tried to summarize the little information available on the nutritional and physicochemical requirements determining the sink strength. Because CO is highly toxic to respiring organisms even in low concentrations, the function of microbial activities in the global CO cycle is critical.
    Materialart: Digitale Medien
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Archives of microbiology 154 (1990), S. 168-174 
    ISSN: 1432-072X
    Schlagwort(e): CO ; Nitrite ; Nitrous oxide ; Nitrogen assimilation ; Carboxydotrophic bacteria ; Pseudomonas carboxydoflava ; Pseudomonas carboxydohydrogena ; Pseudomonas carboxydovorans
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We describe the ability of carboxydotrophic bacteria for nitrate respiration or denitrification. Four out of fourteen strains examined could denitrify heterotrophically forming N2 (Pseudomonas carboxydoflava) or N2O (Pseudomonas carboxydohydrogena, Pseudomonas compransoris, and Pseudomonas gazotropha). Three carried out a heterotrophic nitrate respiration (Arthrobacter 11/x, Azomonas B1, and Azomonas C2). P. carboxydohydrogena could use H2 as electron donor for nitrate respiration under chemolithoautotrophic growth conditions. CO did not support denitrification or nitrate respiration of carboxydotrophic bacteria, although the free energy changes of the reactions would be sufficiently negative to allow growth. CO at 50 kPa was a weak inhibitor of N2O-reduction in carboxydotrophic and non-carboxydotrophic bacteria and decelerated denitrifying growth. Carboxydotrophic bacteria could utilize a wide range of N-sources. Results obtained with a plasmid-cured mutant of Pseudomonas carboxydovorans OM5 showed, that genes involved in nitrogen assimilation entirely reside on the chromosome. In the presence of an suitable electron donor, most carboxydotrophic bacteria could carry out a reduction of nitrate to nitrite that did not support growth and did not lead to the formation of ammonia.
    Materialart: Digitale Medien
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