Identification of a transcriptional activator (ChnR) and a 6-oxohexanoate dehydrogenase (ChnE) in the cyclohexanol catabolic pathway in Acinetobacter sp. Strain NCIMB 9871 and localization of the genes that encode them.
We identified chnR, a gene encoding an AraC-XylS type of transcriptional activator that regulates the expression of chnB, the structural gene for cyclohexanone monooxygenase (CHMO) in Acinetobacter sp. strain NCIMB 9871. The gene sequence of chnE, which encodes an NADP(+)-linked 6-oxohexanoate dehydrogenase, the enzyme catalyzing the fifth step of cyclohexanol degradation, was also determined. The gene arrangement is chnB-chnE-chnR. The predicted molecular masses of the three polypeptides were verified by radiolabeling by using the T7 expression system. Inducible expression of cloned chnB in Escherichia coli depended upon the presence of chnR. A transcriptional chnB::lacZ fusion experiment revealed that cyclohexanone induces chnB expression in E. coli, in which a 22-fold increase in activity was observed.
Iwaki
H
H
Department of Biotechnology, Faculty of Engineering & High Technology Research Center, Kansai University, Suita, Osaka 564-8680, Japan.
Hasegawa
Y
Y
Teraoka
M
M
Tokuyama
T
T
Bergeron
H
H
Lau
P C
PC
D000150:Acinetobacter / Q000201:enzymology* / Q000235:genetics*; D000445:Aldehyde Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D016384:Consensus Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000235:genetics; D015183:Restriction Mapping; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism*
D001426:Bacterial Proteins; D015534:Trans-Activators; C400926:chnR protein, Acinetobacter; D010105:Oxygenases; C019961:cyclohexanone oxygenase; D000445:Aldehyde Oxidoreductases; C400925:6-oxohexanoate dehydrogenase