Cloning and sequencing of a 2,5-dichloro-2,5-cyclohexadiene-1,4-diol dehydrogenase gene involved in the degradation of gamma-hexachlorocyclohexane in Pseudomonas paucimobilis.
In Pseudomonas paucimobilis UT26, gamma-hexachlorocyclohexane (gamma-HCH) is converted to 2,5-dichloro-2,5-cyclohexadiene-1,4-diol (2,5-DDOL), which is then metabolized to 2,5-dichlorohydroquinone. Here, we isolated from the genomic library of UT26 two genes which expressed 2,5-DDOL dehydrogenase activity when they were transformed into P. putida and Escherichia coli. Both gene products had an apparent molecular size of 28 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The first gene, named linC, located separately from the two genes (linA and linB) which we had already cloned as genes involved in the gamma-HCH degradation. The other, named linX, located about 1 kb upstream of the linA gene encoding gamma-HCH dehydrochlorinase. A gamma-HCH degradation-negative mutant, named UT72, which lacked the whole linC gene but had the intact linX gene was isolated. The linC gene given in a plasmid could complement UT72. These results strongly suggest that the linC gene but not the linX gene is essential for the assimilation of gamma-HCH in UT26. Deduced amino acid sequences of LinC and LinX show homology to those of members of the short-chain alcohol dehydrogenase family.
Nagata
Y
Y
Department of Agricultural Chemistry, University of Tokyo, Japan.
Ohtomo
R
R
Miyauchi
K
K
Fukuda
M
M
Yano
K
K
Takagi
M
M
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003511:Cyclohexanols / Q000378:metabolism*; D053138:Cyclohexenes; D004252:DNA Mutational Analysis; D016208:Databases, Factual; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D015698:Genomic Library; D001556:Hexachlorocyclohexane / Q000378:metabolism*; D006873:Hydroquinones / Q000378:metabolism; D008969:Molecular Sequence Data; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid
C087473:2,5-dichloro-2,5-cyclohexadiene-1,4-diol; D003511:Cyclohexanols; D053138:Cyclohexenes; D006873:Hydroquinones; D011994:Recombinant Proteins; D001556:Hexachlorocyclohexane; C069879:2,5-dichlorohydroquinone; C092148:2,5-dichloro-2,5-cyclohexadiene- 1,4-diol dehydrogenase; D000429:Alcohol Oxidoreductases