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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 149 (1987), S. 112-119 
    ISSN: 1432-072X
    Keywords: Methylobacillus flagellatum KT ; Obligate methylotroph ; Nitrosoguanidine mutagenesis ; Auxotrophic mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A collection of polyauxotrophic mutants of the obligate methylotroph Methylobacillus flagellatum KT was obtained. On the first step two stable auxotrophic mutants with a high requirement for amino acids supplements were isolated by treatment with nitrosoguanidine and selection on complete medium. Spontaneous variants of these mutants with a low requirement for nutrient supplements were the base for obtaining polyauxotrophic strains. It was shown, that the growth of mutants of M. flagellatum KT is inhibited by complete medium. Some amino acids and nucleotides are the inhibitor components of complete media. An approach for selection of auxotrophic mutants of individual genes was worked out on minimal medium. The optimal conditions for nitrosoguanidine mutagenesis of M. flagellatum KT were developed. The possible mechanisms of action of some of the nutrient supplements on the growth of M. flagellatum KT are discussed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-072X
    Keywords: Methylobacillus flagellatum ; Obligate methylotroph ; Oxyturbidostat ; Formaldehyde ; C-1 metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ribulose monophosphate cycle methylotroph Methylobacillus flagellatum was grown under oxyturbidostat conditions on mixtures of methanol and formaldehyde. Formaldehyde when added at low concentration (50 mg/l) increased the methanol consumption and the yield of biomass. The presence of 150–300 mg/l of formaldehyde resulted in an increase of the growth rate from 0.74 to about 0.79–0.82 h-1. The presence of 500 mg/l of formaldehyde in the inflow decreased culture growth characteristics. Activities of methanol dehydrogenase and enzymes participating in formaldehyde oxidation and assimilation were measured. The enzymological profiles obtained are discussed.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 153 (1990), S. 139-145 
    ISSN: 1432-072X
    Keywords: Methylobacillus flagellatum ; Obligate methylotroph ; Inc P1 plasmids ; Retrotransfer ; Chromosome mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Homologous matings with plasmids R68.45 and pULB113, and also with Hfr type donor were employed for mapping pgi and gpd genes involved in C-1 metabolism in the obligate methylotroph Methylobacillus flagellatum. A preliminary map of the late chromosomal region was constructed on the basis of these experimental results. The C-1 markers were linked to methionine and leucine auxotrophy and nalidixic acid resistance markers. The phenomenon of retrotransfer, or shuttle transfer of chromosomal markers by Inc P1 plasmids, revealed earlier, was demonstrated for M. flagellatum.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-072X
    Keywords: Methylobacillus flagellatum ; Obligate methylotroph ; Ribulose monophosphate cycle ; Temperature sensitive mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An approach to the isolation of mutants deficient in key enzymes of C1 metabolism was worked out for the obligate methylotroph Methylobacillus flagellatum KT. The isolation of mutants in genes of the ribulose monophosphate (RMP) cycle of formaldehyde assimilation and oxidation is based on selection of temperature sensitive (ts) mutants followed by enzymatic assays. Standard methods of determination of activities of hexulose phosphate synthase/hexulose phosphate isomerase (HPS/HPI), phosphoglucoisomerase (PGI), glucose 6-phosphate dehydrogenase (GPD) and 6-phosphogluconate dehydrogenase (GND) were modified for screening the ts-mutants for the presence of RMP keyenzyme activities. Among the 500 ts-mutants investigated, nine mutants were defective in PGI activity and two in GDP activity. The defective enzymes were characterized by a high rate of inactivation at increased temperatures. At 50° C the proteins studied were completely inactivated during 2 h, whereas the native enzymes maintained more than 80% activity.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 258 (1998), S. 133-138 
    ISSN: 1617-4623
    Keywords: Key words Hfr donor ; Mapping ; Obligate methylotroph ; Methylobacillus flagellatum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We present a refined genetic map of the obligate methylotroph Methylobacillus flagellatum. New, Hfr (high-frequency-of-transfer) donors, and pulsed-field gel electrophoresis, were used to determine that M.␣flagellatum contains one ∼3.1-Mb circular chromosome, and no plasmids. A correlation between time-of-entry units and DNA length was established. Using in vivo and in vitro cloning, and sequencing, a number of new genetic markers were identified and mapped; in addition, the nature of some of the previously mapped markers was elucidated.
    Type of Medium: Electronic Resource
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