Molecular cloning of a Pseudomonas paucimobilis gene encoding a 17-kilodalton polypeptide that eliminates HCl molecules from gamma-hexachlorocyclohexane.
Pseudomonas paucimobilis UT26 is capable of growing on gamma-hexachlorocyclohexane (gamma-HCH). A genomic library of P. paucimobilis UT26 was constructed in Pseudomonas putida by using the broad-host-range cosmid vector pKS13. After 2,300 clones were screened by gas chromatography, 3 clones showing gamma-HCH degradation were detected. A 5-kb fragment from one of the cosmid clones was subcloned into pUC118, and subsequent deletion and gas chromatography-mass spectrometry analyses revealed that a fragment of ca. 500 bp was responsible for the conversion of gamma-HCH to 1,2,4-trichlorobenzene via gamma-pentachlorocyclohexene. Nucleotide sequence analysis revealed an open reading frame (linA) of 465 bp within the fragment. The nucleotide sequence of the linA gene and the deduced amino acid sequence showed no similarity to any known sequences. The product of the linA gene was 16.5 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Imai
R
R
Department of Agricultural Chemistry, University of Tokyo, Japan.
Nagata
Y
Y
Fukuda
M
M
Takagi
M
M
Yano
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003062:Codon; D004269:DNA, Bacterial; D005798:Genes, Bacterial; D001556:Hexachlorocyclohexane / Q000031:analogs & derivatives / Q000378:metabolism*; D006851:Hydrochloric Acid / Q000378:metabolism*; D008190:Lyases / Q000235:genetics* / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D015183:Restriction Mapping
D001426:Bacterial Proteins; D003062:Codon; D004269:DNA, Bacterial; C010275:pentachlorocyclohexene; D001556:Hexachlorocyclohexane; D008190:Lyases; C019994:dehydrochlorinases; C071445:linA protein, Pseudomonas paucimobilis; D006851:Hydrochloric Acid