Methane monooxygenase mutants ofMethylosinus trichosporiumconstructed by marker-exchange mutagenesis

• DOI: 10.1111/j.1574-6968.1995.tb07480.x
• Publication Date: 2006-01-17T10:09:51Z
• Authors: H Martin; J.C Murrell
• Keywords: Methane Monooxygenase; Methanotroph

Methylosinus trichosporium OB3b synthesizes a soluble cytoplasmic methane monooxygenase when grown in copper-depleted medium and membrane-bound particulate under copper-replete conditions. The genes encoding the hydroxylase component of monooxygenase, carried on plasmid Escherichia coli, were insertionally inactivated using kanamycin cassette transferred back into M. by conjugation. Marker-exchange mutagenesis, via double homologous recombination event, yielded monooxygenase-negative mutant which grew only during growth conditions, thus proving that two oxidation systems this methanotroph are genetically distinct.