Characterization of the Rhodococcus sp. NI86/21 gene encoding alcohol: N,N'-dimethyl-4-nitrosoaniline oxidoreductase inducible by atrazine and thiocarbamate herbicides.
A protein with a mol.mass of 51,000 (ThcE) that was induced in Rhodococcus sp. NI86/21 during assimilation of thiocarbamate herbicides, atrazine, ethanol, propanol, glycerol, propionaldehyde or ethanolamine was identified by two-dimensional electrophoresis. The thcE gene was cloned and sequenced. The deduced amino acid sequence revealed ThcE as a member of group III alcohol dehydrogenases. ThcE displayed strong homology with sequenced subunit fragments of the homodecameric N,N'-dimethyl-4-nitrosoaniline-dependent alcohol oxidoreductases (MNO) of Amycolatopsis methanolica and Mycobacterium gastri. N-Terminal sequence analysis of purified MNO from Rhodococcus sp. NI86/21 confirmed the identity with ThcE. When overproduced in Escherichia coli, ThcE was insoluble and no MNO activity was detected.
Nagy
I
I
F. A. Janssens Laboratory of Genetics, Catholic University of Leuven, Heverlee, Belgium.
Verheijen
S
S
De Schrijver
A
A
Van Damme
J
J
Proost
P
P
Schoofs
G
G
Vanderleyden
J
J
De Mot
R
R
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D000595:Amino Acid Sequence; D001280:Atrazine / Q000494:pharmacology*; D001483:Base Sequence; D002219:Carbamates; D003001:Cloning, Molecular; D004790:Enzyme Induction; D000431:Ethanol / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005798:Genes, Bacterial / Q000235:genetics*; D006540:Herbicides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D012240:Rhodococcus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D002219:Carbamates; D006540:Herbicides; D000431:Ethanol; D000429:Alcohol Oxidoreductases; C095705:alcohol - N, N'-dimethyl-4-nitrosoaniline oxidoreductase; D001280:Atrazine