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Identification of putative methanol dehydrogenase (moxF) structural genes in methylotrophs and cloning of moxF genes from methylococcus capsulatus bath and Methylomonas albus BG8

Journal Article · · J. Bacteriol.; (United States)
OSTI ID:6769422
; ;
An open-reading-frame fragment of a Methylobacterium sp. strain AM1 gene (moxF) encoding a portion of the methanol dehydrogenase structural protein has been used as a hybridization probe to detect similar sequences in a variety of methylotrophic bacteria. This hybridization was used to isolate clones containing putative moxF genes from two obligate methanotrophic bacteria, Methylococcus capsulatus Bath and Methylomonas albus BG8. The identity of these genes was confirmed in two ways. A T7 expression vector was used to produce methanol dehydrogenase protein in Escherichia coli from the cloned genes,a and in each case the protein was identified by immunoblotting with antiserum against the Methylomonas albus methanol dehydrogenase. In addition, a moxF mutant of Methylobacterium strain AM1 was complemented to a methanol-positive phenotype that partially restored methanol dehydrogenase activity, using broad-host-range plasmids containing the moxF genes from each methanotroph. The partial complementation of a moxF mutant in a facultative serine pathway methanol utilizer by moxF genes from type I and type X obligate methane utilizers suggests broad functional conservation of the methanol oxidation system among gram-negative methylotrophs.
Research Organization:
California Institute of Technology, Pasadena (USA)
OSTI ID:
6769422
Journal Information:
J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 170:5; ISSN JOBAA
Country of Publication:
United States
Language:
English

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Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ALCOHOL DEHYDROGENASE
ALCOHOLS
BACTERIA
CELL CONSTITUENTS
CHEMICAL REACTIONS
CLONING
DNA-CLONING
ENZYMES
GENES
HEMIACETAL DEHYDROGENASES
HYBRIDIZATION
HYDROXY COMPOUNDS
METHANOL
MICROORGANISMS
MUTANTS
ORGANIC COMPOUNDS
OXIDATION
OXIDOREDUCTASES
PLASMIDS