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  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Archives of microbiology 171 (1999), S. 417-423 
    ISSN: 1432-072X
    Schlagwort(e): Key words Alanine dehydrogenase ; Ammonia ; assimilation ; Mycobacterium ; Morpholine degradation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract An NAD-dependent, morpholine-stimulated l-alanine dehydrogenase activity was detected in crude extracts from morpholine-, pyrrolidine-, and piperidine-grown cells of Mycobacterium strain HE5. Addition of morpholine to the assay mixture resulted in an up to 4.6-fold increase of l-alanine dehydrogenase activity when l-alanine was supplied at suboptimal concentration. l-Alanine dehydrogenase was purified to near homogeneity using a four-step purification procedure. The native enzyme had a molecular mass of 160 kDa and contained one type of subunit with a molecular mass of 41 kDa, indicating a tetrameric structure. The sequence of 30 N-terminal amino acids was determined and showed a similarity of up to 81% to that of various alanine dehydrogenases. The pH optimum for the oxidative deamination of l-alanine, the only amino acid converted by the enzyme, was determined to be pH 10.1, and apparent K m values for l-alanine and NAD were 1.0 and 0.2 mM, respectively. K m values of 0.6, 0.02, and 72 mM for pyruvate, NADH, and NH4 +, respectively, were estimated at pH 8.7 for the reductive amination reaction.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1432-072X
    Schlagwort(e): Interspecies hydrogen transfer ; Interspecies formate transfer ; Alanine ; Serine ; Glycine fermentation ; Selenium ; Glycine reductase ; Sarcosine reduction ; Betaine reduction ; Eubacterium acidaminophilum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract An obligately anaerobic, rod-shaped bacterium was isolated on alanine in co-culture with H2-scavenging Desulfovibrio and obtained in pure culture with glycine as sole fermentation substrate. The isolated strain, al-2, was motile by a polar to subpolar flagellum and stained Gram-positive. The guanine plus cytosine content of the DNA was 44.0 mol%. Strain al-2 grew in defined, reduced glycine media supplemented with biotin. The pure culture fermented 4 mol glycine to 3 mol acetate, 4 mol ammonia and 2 mol CO2. Under optimum conditions (34°C, pH 7.3), the doubling time on glycine was 60 min and the molar growth yield 7.6 g cell dry mass. Serine was fermented to acetate, ethanol, CO2, H2 and ammonia. In addition, betaine, sarcosine or creatine served as substrates for growth and acetate production if H2, formate or e.g. valine were added as H-donors. In pure culture on alanine under N2, strain al-2 grew very poorly and produced H2 up to a partial pressure of 3.6 kPa (0.035 atm). Desulfovibrio species, Methanospirillum hungatei and Acetobacterium woodii served as H2-scavengers that allowed good syntrophic growth on alanine. The co-cultures also grew on aspartate, leucine, valine or malate. Alanine and aspartate were stoichiometrically degraded to acetate and ammonia, whereas the reducing equivalents were recovered as H2S, CH4 or newly synthetized acetate, respectively. Growth of strain al-2 in co-culture with the hydrogenase-negative, formate-utilizing Desulfovibrio baarsii indicated that a syntrophy was also possible by interspecies formate transfer. Growth on glycine, or on betaine, sarcosine or creatine (plus H-donors) depended strictly on the addition of selenite (≥0.1 μM); selenite was not required for fermentation of serine, or for degradation of alanine, aspartate or valine by the co-cultures. Cell-free extracts of glycine-grown cells contained active glycine reductase, glycine decarboxylase and reversible methyl viologen-dependent formate dehydrogenase in addition to the other enzymes necessary for an oxidation to CO2. In all reactions NADP was the preferred H-carrier. Both formate and glycine could be synthesized from bicarbonate. Serine-grown cells did not contain serine hydroxymethyl transferase but serine dehydratase and other enzymes commonly involved in pyruvate metabolism to acetate, CO2 and H2. The enzymes involved in glycine metabolism were repressed during growth on serine. By its morphology and physiology, strain al-2 did not resemble described amino acid-degrading species. Therefore, the new isolate is proposed as type strain of a new species, Eubacterium acidaminophilum.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Archives of microbiology 134 (1983), S. 167-169 
    ISSN: 1432-072X
    Schlagwort(e): Nitrogenase ; Glutamine synthetase ; Glutamate synthase ; Glutamate dehydrogenase ; Asparagine synthetase ; Alanine dehydrogenase ; β-Methylaspartase ; Clostridium formicoaceticum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Clostridium formicoaceticum possesses the following enzymes for the assimilation of N2 and NH 4 + : nitrogenase, glutamine synthetase, NADH- and NADPH-dependent glutamate synthase, NADH- and NADPH-dependent glutamate dehydrogenase, NADPH-dependent alamine dehydrogenase, and NH 4 + -dependent asparagine synthetase. Nitrogenase and glutamine synthetase are repressed and alanine dehydrogenase is induced by NH 4 + , while the synthesis of the other enzymes is not influenced by the extracellular NH 4 + level. Glutamate is degraded via glutamate mutase and β-methylaspartase.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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