Identification and functional characterization of a gene for the methanol : N,N'-dimethyl-4-nitrosoaniline oxidoreductase from Mycobacterium sp. strain JC1 (DSM 3803).
Mycobacterium sp. strain JC1 is able to grow on methanol as a sole source of carbon and energy using methanol : N,N'-dimethyl-4-nitrosoaniline oxidoreductase (MDO) as a key enzyme for primary methanol oxidation. Purified MDO oxidizes ethanol and formaldehyde as well as methanol. The Mycobacterium sp. strain JC1 gene for MDO (mdo) was cloned, sequenced, and determined to have an open reading frame of 1272 bp. Northern blot and promoter analysis revealed that mdo transcription was induced in cells grown in the presence of methanol. Northern blotting together with RT-PCR also showed that the mdo gene was transcribed as monocistronic mRNA. Primer extension analysis revealed that the transcriptional start site of the mdo gene is located 21 bp upstream of the mdo start codon. An mdo-deficient mutant of Mycobacterium sp. strain JC1 did not grow with methanol as a sole source of carbon and energy.
Park
Hyuk
H
Molecular Microbiology Laboratory, Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea.
Lee
Hyunil
H
Laboratory of Biochemistry, Graduate School of Medicine, Konkuk University, Seoul 143-701, Republic of Korea.
Ro
Young T
YT
Laboratory of Biochemistry, Graduate School of Medicine, Konkuk University, Seoul 143-701, Republic of Korea.
Kim
Young M
YM
Molecular Microbiology Laboratory, Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea.
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D002244:Carbon / Q000378:metabolism; D002384:Catalysis; D003001:Cloning, Molecular; D004734:Energy Metabolism; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009161:Mycobacterium / Q000201:enzymology* / Q000235:genetics; D012329:RNA, Bacterial; D012333:RNA, Messenger; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D002244:Carbon; D000429:Alcohol Oxidoreductases; C095705:alcohol - N, N'-dimethyl-4-nitrosoaniline oxidoreductase; D005947:Glucose; D000432:Methanol