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  • 1
    Electronic Resource
    Electronic Resource
    Mutants of the obligate methylothroph Methylobacillus flagellatum KT defective in genes of the ribulose monophosphate cycle of formaldehyde fixation (1988)
    Springer
    Archives of microbiology 149 (1988), S. 441-446 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00425585
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  • 2
    Electronic Resource
    Electronic Resource
    Development of gene transfer systems in Methylobacillus flagellatum KT: Isolation of auxotrophic mutants (1987)
    Springer
    Archives of microbiology 149 (1987), S. 112-119 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00425075
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  • 3
    Electronic Resource
    Electronic Resource
    Mapping of pgi and gpd genes involved in C-1 assimilation in the obligate methylotroph Methylobacillus flagellatum (1990)
    Springer
    Archives of microbiology 153 (1990), S. 139-145 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00247811
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  • 4
    Electronic Resource
    Electronic Resource
    Effect of formaldehyde on growth of obligate methylotroph Methylobacillus flagellatum in a substrate non-limited continuous culture (1992)
    Springer
    Archives of microbiology 158 (1992), S. 145-148 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00245218
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  • 5
    Electronic Resource
    Electronic Resource
    2H- and13C-labeled amino acids generated by obligate methylotrophs Biosynthesis and MS monitoring (1994)
    Springer
    Amino acids 6 (1994), S. 165-176 
    Details
    ISSN: 1438-2199
    Keywords: Amino acids ; Stable isotope labeled amino acids ; Biosynthesis ; Methylobacillus flagellatum ; Electron impact mass spectrometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Biosynthetic preparation of2H- and13C- labeled amino acids was studied using a leucine-producing mutant of the obligate methylotroph,Methylobacillus flagellatum. The strain was cultivated in various media containing13C- or2H-analogs of methanol. The total protein from each experiment was subjected to acid hydrolysis and converted into a mixture of dansyl amino acid methyl esters. The samples of excreted leucine were converted into methyl esters of dansyl and benzyloxycarbonyl derivatives. Electron impact mass spectrometry was performed to detect stable isotope enrichment of the amino acids. According to the mass spectrometric analysis it is feasible to use methylotrophic microorganisms for the preparation of2H- and13C- analogs of amino acids by labeled methanol bioconversion; the excreted amino acids can be convenient for express analysis as an indicator of isotopic enrichment of the total protein. The data obtained testified to a high efficiency of dansyl derivatization for mass spectrometric analysis of complex amino acid mixtures.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00805844
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  • 6
    Electronic Resource
    Electronic Resource
    Oxidative and assimilative enzyme activities in continuous cultures of the obligate methylotrophMethylobacillus flagellatum (1991)
    Springer
    Antonie van Leeuwenhoek 60 (1991), S. 101-107 
    Details
    ISSN: 1572-9699
    Keywords: continuous culture ; enzyme levels ; Methylobacillus flagellatum ; obligate methylotroph
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In methanol-limited continuous cultures of the obligate methylotrophic bacteriumMethylobacillus flagellatum grown at rates from 0.05 to 0.63 h-1, and also in an oxyturbidostat culture ofM. flagellatum growing at the rate of 0.73 h-1, levels of methanol dehydrogenase, enzymes of formaldehyde oxidation (both linear and cyclic) and assimilation (RuMP cycle), a number of intermediary metabolism and TCA cycle enzymes and also ‘dye-linked’ formaldehyde dehydrogenase were determined. It was shown that the activities of dissimilatory enzymes, with the exception of ‘dye-linked’ formaldehyde dehydrogenase, decreased with increasing growth rate. Activities of assimilative enzymes and activities of the TCA cycle enzymes detected as well as the ‘dye-linked’ formaldehyde dehydrogenase activity, increased with increasing growth rate. A periplasmic location was shown for the latter enzyme and a role in formaldehyde detoxification was proposed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00572699
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  • 7
    Electronic Resource
    Electronic Resource
    Growth of the obligate methylotroph Methylobacillus flagellatum under stationary and nonstationary conditions during continuous cultivation (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 39 (1992), S. 688-695 
    Details
    ISSN: 0006-3592
    Keywords: methylotroph ; continuous culture ; oxiturbidostat ; growth-limiting substrate pulse ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The growth characteristics of a chemostat culture of the obligate methylotrophic bacterium Methylobacillus flagellatum have been determined. Steady-state cultures growing at a rate of 0.73-0.74 h-1, equal to the maximal growth rate, were obtained under oxyturbidostat cultivation conditions. The response of a chemostat culture to a pulse increase of methanol concentration was studied. It was shown that slow and rapidly growing cultures of M. flagellatum responded differently to pulse methanol addition. The growth characteristics of slow-growing cultures decreased after methanol addition compared to those of stationary chemostat cultures. The growth characteristics of rapidly growing cultures were practically unchanged with and without pulse methanol addition.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260390614
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  • 8
    Electronic Resource
    Electronic Resource
    Refined genetic map of the obligate methylotroph Methylobacillus flagellatum (1998)
    Springer
    Molecular genetics and genomics 258 (1998), S. 133-138 
    Details
    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
    URL: http://dx.doi.org/10.1007/s004380050715
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  • 9
    Electronic Resource
    Electronic Resource
    Construction of a promoter-probe vector for the methanol-utilizing bacterium acetobacter methanolicus MB 58 (1989)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 9 (1989), S. 219-225 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: We report here the construction of a promoter-probe vector, pRS2, which can be utilized in either Acetobacter methanolicus MB 58 or Escherichia coli due to the presence of broad-host-range replicon RSF 1010. The vector provides several unique restriction sites for promoter cloning as well as resistance markers for the selection of transformants. The promoter-probe vector was constructed by inserting an EcoRI-SalI-polylinker fragment of pUC 19 into EcoRI/SalI digested pMK 16. The resulting plasmid, pRS1, was cloned into the unique EcoRI site of the broad-host-range plasmid RSF 1010.The vector was used to clone promoter-containing sequences derived from the A. methanolicus MB 58 chromosome as well as the E. coli lac-promoter.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370090306
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