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  • 1
    ISSN: 1432-072X
    Keywords: Methanotroph ; Methane oxidation ; Methane monooxygenase ; Oligonucleotide probing ; Subunit genes ; Methylococcus capsulatus (Bath)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methane monooxygenase (MMO) is the enzyme responsible for the conversion of methane to methanol in methanotrophic bacteria. In addition, this enzyme complex oxidizes a wide range of aliphatic and aromatic compounds in a number of potentially useful biotransformations. In this study, we have used biochemical data obtained from purification and characterization of the soluble MMO from Methylococcus capsulatus (Bath), to identify structural genes encoding this enzyme by oligonucleotide probing. The genes encoding the β and γ subunits of MMO were found to be chromosomally located and were linked in this organism. We report here on the analysis of a recombinant plasmid containing 12 kilobases of Methylococcus DNA and provide the first evidence for the localization and linkage of genes encoding the methane monooxygenase enzyme complex. DNA sequence analysis suggests that the primary structures of the β and γ subunit of MMO are completely novel and the complete sequence of these genes is presented.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Biodegradation 5 (1994), S. 145-159 
    ISSN: 1572-9729
    Keywords: methane oxidation ; methane monooxygenase gene ; Methylococcus capsulatus (Bath) ; Methylosinus trichosporium OB3b
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Biological methane oxidation is carried out by methanotrophs, bacteria that utilize methane as their sole carbon and energy source. The enzyme they contain that is responsible for methane oxidation is methane monooxygenase, the most well studied being the soluble methane monooxygenase enzyme complexes fromMethylococcus capsulatus (Bath) andMethylosinus trichosporium OB3b. In both organisms, the genes encoding soluble methane monooxygenase have been found to be clustered on the chromosome in the ordermmoX, mmoY, mmoB, mmoZ, orfY andmmoC. These genes encode the α and β subunits of Protein A, Protein B, the γ subunit of Protein A, a protein of unknown function and Protein C respectively of the soluble methane monooxygenase complex. The complete DNA sequences of both gene clusters have been determined and they show considerable homology. Expression of soluble methane monooxygenase genes occurs under growth conditions where the copper-to-biomass ratio is low. Transcriptional regulation of the gene cluster fromMethylosinus occurred at an RpoN-like promoter, 5′ of themmoX gene.mmoB andmmoC ofMethylococcus have been expressed inE. coli and the proteins obtained were functionally active. Soluble methane monooxygenase mutants have been constructed by marker-exchange mutagenesis. They were found to be more stable than those generated using the suicide substrate dichloromethane. Soluble methane monooxygenase probes have been used to detect both methane monooxygenase gene-specific DNA and methanotrophs in natural environmental samples.
    Type of Medium: Electronic Resource
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