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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Macmillan Magazines Ltd.
    Nature 406 (2000), S. 37-37 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Biological phosphorus occurs almost exclusively as phosphate in the redox state of + V, although a few phosphonic (+ III) and phosphinic (+ I) acids are found as secondary metabolites or as constituents of phosphonolipids. Here we show that a culture of a lithoautotrophic bacterium ...
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology reviews 15 (1994), S. 0 
    ISSN: 1574-6976
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract: Fatty acids are key intermediates in methanogenic degradation of organic matter in sediments as well as in anaerobic reactors. Conversion of butyrate or propionate to acetate, (CO2), and hydrogen is endergonic under standard conditions, and becomes possible only at low hydrogen concentrations (10-4-10-5 bar). A model of energy sharing between fermenting and methanogenic bacteria attributes a maximum amount of about 20 kJ per mol reaction to each partner in this syntrophic cooperation system. This amount corresponds to synthesis of only a fraction (one-third) of an ATP to be synthesized per reaction. Recent studies on the biochemistry of syntrophic fatty acid-oxidizing bacteria have revealed that hydrogen release from butyrate by these bacteria is inhibited by a protonophore or the ATPase inhibitor DCCD (N,N′-dicyclohexyl carbodiimide), indicating that a reversed electron transport step is involved in butyrate or propionate oxidation. Hydrogenase, butyryl-CoA dehydrogenase, and succinate dehydrogenase acitivities were found to be partially associated with the cytoplasmic membrane fraction. Also glycolic acid is degraded to methane and CO2 by a defined syntrophic coculture. Here the most difficult step for hydrogen release is the glycolate dehydrogenase reaction (E′0=−92 mV). Glycolate dehydrogenase, hydrogenase, and ATPase were found to be membrane-bound enzymes. Membrane vesicles produced hydrogen from glycolate only in the presence of ATP; protonophores and DCCD inhibited this hydrogen release. This system provides a suitable model to study reversed electron transport in interspecies hydrogen transfer between fermenting and methanogenic bacteria in methanogenic biomass degradation.
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  • 3
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Earthworms are important members of the soil macrofauna. They modify soil physical properties, soil organic matter decomposition, and thus regulate carbon and nitrogen cycling in soil. However, their interactions with soil microorganisms are still poorly understood, in particular the effect of gut passage on the community structure of ingested microorganisms. Moreover, it is still unsolved, if earthworms, like many other soil-feeding invertebrates, possess an indigenous gut microbial community. Therefore, we investigated the bacterial and archaeal community structure in soil (with and without additional beech litter), gut, and fresh casts of Lumbricus terrestris, an anecic litter-feeding earthworm, by means of terminal-restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA gene fragments. Ecological indices of community diversity and similarity, calculated from the T-RFLP profiles, revealed only small differences between the bacterial and archaeal communities in soil, gut, and fresh casts under both feeding conditions, especially in comparison to other soil-feeding invertebrates. However, multivariate statistical analysis combining multidimensional scaling and discriminant function analysis proved that these differences were highly significant, in particular when the earthworms were fed beech litter in addition. Because there were no dominant gut-specific OTUs detectable, the existence of an abundant indigenous earthworm microbial community appears unlikely, at least in the midgut region of L. terrestris.
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 38 (2001), S. 0 
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Addition of straw to anoxic rice field soil stimulates production of CH4, an important greenhouse gas. The archaeal community colonizing rice straw was investigated by molecular methods targeting the small subunit ribosomal RNA gene. Cloning and sequencing of 60 clones detected predominantly relatives of Methanobacterium spp. (38 clones) and Methanosarcina spp. (16 clones). Terminal restriction fragment length polymorphism (T-RFLP) analysis confirmed the dominance of Methanobacteriaceae and Methanosarcinaceae, and in addition showed restriction fragments characteristic for Rice cluster I (RC-I) methanogens. A new oligonucleotide probe specific for RC-I was designed. Quantitative slot blot hybridization of extracted rRNA with this probe indicated the presence of an active population of RC-I methanogens. Other methanogenic groups (e.g. Methanomicrobiaceae, Methanosaetaceae), although present and active in soil, could not be conclusively detected on rice straw. The methanogenic community pattern on straw, as revealed by T-RFLP and quantitative rRNA probing, was fairly constant with incubation time (8–57 days), but the total activity of methanogenic Archaea almost doubled. Our results indicate that the methanogens colonizing rice straw are less diverse than those inhabiting the soil.
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  • 5
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The methane production potential of rice soils, which are situated in different geographical regions, shows inherent variations and is catalyzed by archaeal methanogens. We therefore investigated the archaeal community structure in 11 rice field soils which represent a range of climatic conditions (temperate to subtropical zones) and soil properties. Retrieval of environmental partial SSU rDNA sequences from the rice soils of Shenyang (China) and Gapan (The Philippines) showed that the communities were different from each other. However, despite the differences in soil properties and geographical region the sequences clustered in similar phylogenetic groups to those obtained earlier from rice fields of Vercelli (Italy). The archaeal community structure in the other rice field soils was compared using terminal restriction fragment length polymorphism (T-RFLP) analysis targeting the SSU rRNA gene and the methyl-coenzyme M reductase α-subunit gene (mcrA). The relative abundance of each terminal restriction fragment (T-RF) was determined by fluorescence peak area integration. The 182-bp SSU rDNA T-RF (representing members of Methanosarcinaceae and rice cluster (RC) VI) was dominant (40–80% contribution) in Chinese soils (Zhenjiang, Changchun, Jurong, Beiyuan, Shenyang) and the Philippine soil of Gapan. The other Philippine soils (Luisiana, Guangzhou, Pila) and the Italian soils (Vercelli, Pavia) showed a dominant 389-bp T-RF (35–40% contribution), representing mainly the novel methanogenic RC-I. All the other T-RF (80, 88, 280, 375 and 〉800 bp) contributed 〈20%. Prolonged anoxic incubation (30–200 days) of the air-dried soils resulted in the production of CH4, which was in some soils preceded by a characteristic halt phase. T-RFLP analysis revealed that the soils with a methanogenic halt phase also showed dramatic archaeal population dynamics which were related to the length of the halt phase. Our results show that the archaeal communities in rice field soils of different geographical origin are highly related, but nevertheless exhibit individual patterns and dynamics, thus providing evidence for the active participation of the community members in energy and carbon flow.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 156 (1991), S. 392-397 
    ISSN: 1432-072X
    Keywords: Glyoxylate ; Anaerobic degradation ; Malic enzyme ; Substrate level phosphorylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A new strictly anaerobic, gram-negative, nonsporeforming bacterium, Strain PerGlx1, was enriched and isolated from marine sediment samples with glyoxylate as sole carbon and energy source. The guanineplus-cytosine content of the DNA was 44.1±0.2 mol %. Glyoxylate was utilized as the only substrate and was stoichiometrically degraded to carbon dioxide, hydrogen, and glycolate. An acetyl-CoA and ADP-dependent glyoxylate converting enzyme activity, malic enzyme, and pyruvate synthase were found at activities sufficient for growth (≥0.25 U x mg protein-1). These findings allow to design a new degradation pathway for glyoxylate: glyoxylate is condensed with acetyl-CoA to form malyl-CoA; the free energy of the thioester linkage in malyl-CoA is conserved by substrate level phosphorylation. Part of the electrons released during glyoxylate oxidation to CO2 reduce a small fraction of glyoxylate to glycolate.
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  • 7
    ISSN: 1432-072X
    Keywords: Electron transport phosphorylation ; Respiration ; Membrane vesicles ; Anaerobic degradation ; Glyoxylate ; Glycolate ; Hydrogen ; Syntrophy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The syntrophically glycolate-fermenting bacterium in the methanogenic binary coculture FlGlyM was isolated in pure culture (strain FlGlyR) with glyoxylate as sole substrate. This strain disproportionated 12 glyoxylate to 7 glycolate, 10 CO2, and 3 hydrogen. Glyoxylate was oxidized via the malyl-CoA pathway. All enzymes of this pathway, i.e. malyl-CoA lyase/malate: CoA ligase, malic enzyme, and pyruvate synthase, were demonstrated in cell-free extracts. Glycolate dehydrogenase, hydrogenase, and ATPase, as well as menaquinones as potential electron carriers, were present in the membranes. Everted membrane vesicles catalyzed hydrogen-dependent glyoxylate reduction to glycolate [86–207 nmol min-1 (mg protein)-1] coupled to ATP synthesis from ADP and Pi [38–82 nmol min-1 (mg protein)-1]. ATP synthesis was abolished entirely by protonophores or ATPase inhibitors (up to 98 and 94% inhibition, respectively) indicating the involvement of proton-motive force in an electron transport phosphorylation driven by a new glyoxylate respiration with hydrogen as electron donor. Measured reaction rates in vesicle preparations revealed a stoichiometry of ATP formation of 0.2–0.5 ATP per glyoxylate reduced.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 156 (1991), S. 398-404 
    ISSN: 1432-072X
    Keywords: Interspecies hydrogen transfer ; Methanogenesis ; Homoacetogenesis ; Malic enzyme ; Substrate level phosphorylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three different defined cocultures of glycolatedegrading strictly anaerobic bacteria were isolated from enrichment cultures inoculated with freshwater sediment samples. Each culture contained a primary fermenting bacterium which used only glycolate as growth substrate. These cells were gram-positive, formed terminal oval spores, and did not contain cytochromes. Growth with glycolate was possible only in coculture with either a homoacetogenic bacterium or a hydrogen-utilizing methanogenic bacterium; the overall fermentation balance was either 4 glycolate → 3 acetate + 2CO2, or 4 glycolate → 3 CH4+5 CO2. These transformations indicate that glycolate was converted by the primary fermenting bacterium entirely to CO2 and reducing equivalents which were transferred to the partner organisms, probably through interspecies hydrogen transfer. The key enzymes of fermentative glycolate degradation were identified in cell-free extracts. An acetyl-CoA and ADP-dependent glyoxylate-converting enzyme activity, malic enzyme, pyruvate synthase, and methyl viologen-dependent hydrogenase were found at comparably high activities suggesting that these bacteria oxidize glycolate through a new pathway via malyl-CoA, and that ATP is synthesized by substrate-level phosphorylation, in a similar manner as found in a recently isolated glyoxylatefermenting anaerobe.
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  • 9
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The human Mut-S-Homologon-2 (hMSH-2) gene product is a member of a highly conserved family of proteins involved in postreplication mismatch repair. We have analysed hMSH-2 expression in normal ovarian tissue (n=15) and ovarian carcinomas (n=40). hMSH-2 protein was investigated immunohistochemically on frozen sections using a highly sensitive streptavidin–peroxidase technique and a specific mouse monoclonal antibody (clone FE11). A hMSH-2-immunoreactivity score (hMSH-2-IRS) for semiquantitative analysis of hMSH-2 expression is presented. In normal ovarian tissue, we only found weak nuclear immunoreactivity for hMSH-2 in 60%, while the remaining 40% were hMSH-2 negative (mean hMSH-2-IRS: 0.73; SD: ±0.70). All ovarian carcinomas analysed revealed moderate to strong nuclear immunoreactivity (mean hMSH-2-IRS: 8.05; SD: ±3.65). hMSH-2 staining was heterogeneous, with visual differences between individual tumour cells. Expression of hMSH-2 protein was consistently and strongly upregulated in tumour cells of ovarian carcinomas as compared to normal ovarian tissue. No statistically significant correlation in comparing the labelling patterns for hMSH-2 with the labelling patterns for Ki-67 (mean percentage of Ki-67 positive tumour cells: 25.88%; SD: ±18.43) was observed in ovarian carcinomas. Furthermore, no statistical significant correlations between hMSH-2-IRS and histological grading (p=0.47), histological type of carcinoma (p=0.706) or FIGO-classification (p=0.054) were found. Our findings indicate that (a) hMSH-2 is expressed in normal human ovarian tissue, (b) expression of hMSH-2 is increased in ovarian carcinomas, (c) expression of hMSH-2 may be of importance for the genetic stability of ovarian carcinomas in vivo, (d) hMSH-2 mutations may not cause microsatellite instability in ovarian carcinomas, (e) hMSH-2 may contribute to mechanisms responsible for resistance to anticancer drugs.
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  • 10
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization and expression of transglutaminase K has been investigated immunohistochemically in normal cervix tissue (n=15) and in cervix carcinomas (n=23). The distribution of the transglutaminase K was compared with the staining patterns of cytokeratin 10, Ki-67, p53, and oestrogen and progesterone receptors in these tumours. Weak to strong membrane-bound immunoreactivity for transglutaminase K was detected in almost all cervix carcinomas analyzed. The immunostaining was heterogeneous, with visual differences between individual tumour cells. 66.7% of normal cervix tissues revealed no immunoreactivity for the transglutaminase K. In normal cervix tissue, the immunoreactivity was confined to upper cervix layers, predominantly to the superficial and intermediate cell layers. The intensity of both the immunostaining and the number of transglutaminase K-positive cells were upregulated in cervix carcinomas as compared to normal cervix tissue. When the coexpressions of transglutaminase K with markers of proliferation and differentiation were analyzed, no statistically significant correlation was found. Our findings indicate that (1) transglutaminase K is upregulated at the protein level in cervix carcinomas as compared to normal cervix tissue; (2) upregulation of the transglutaminase K in cervix carcinoma is not exclusively induced by alterations of epithelial differentiation or proliferation, but by different, unknown mechanisms; and (3) upregulation of transglutaminase K in cervix carcinomas may play an important role for the regulation of tumour invasive properties by modulating cell–cell interactions.
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