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

• Published in: Journal of Bacteriology Vol. 176; no. 11; pp. 3117 - 3125
• Main Authors: NAGATA, Y, OHTOMO, R, MIYAUCHI, K, FUKUDA, M, YANO, K, TAKAGI, M
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.06.1994
• Subjects: alcohol dehydrogenase; alcohol deshidrogenasa; Alcohol Oxidoreductases - biosynthesis; Alcohol Oxidoreductases - genetics; alcool deshydrogenase; Amino Acid Sequence; Base Sequence; biodegradacion; biodegradation; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Blotting, Southern; clonacion; clonage; cloning; Cloning, Molecular; Cyclohexanols - metabolism; Cyclohexenes; Databases, Factual; DNA Mutational Analysis; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; gene; genes; Genes, Bacterial - genetics; genetic transformation; genetica; Genetics; genetique; Genomic Library; hch; Hexachlorocyclohexane - metabolism; Hydroquinones - metabolism; Mission oriented research; Molecular Sequence Data; nucleotide sequence; pseudomonas; Pseudomonas - enzymology; Pseudomonas - genetics; Pseudomonas - metabolism; Pseudomonas paucimobilis; Recombinant Proteins - biosynthesis; Restriction Mapping; secuencia nucleica; Sequence Analysis, DNA; Sequence Deletion; Sequence Homology, Amino Acid; sequence nucleique; transformacion genetica; transformation genetique; Pseudomonadales; Biodegradation; Insecticide; Nucleotide sequence; Homology; Gene organization; Gene expression; HCH; Metabolism; Enzymatic activity; Organochlorine compounds; Bacteria; Pseudomonadaceae; Aminoacid sequence; Structural analysis
• ISSN: 0021-9193; 1098-5530; 1067-8832
• DOI: 10.1128/jb.176.11.3117-3125.1994

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.