Plasmid dependence of Pseudomonas sp. strain NK87 enzymes that degrade 6-aminohexanoate-cyclic dimer.
A bacterial strain, Pseudomonas sp. strain NK87, that can use 6-aminohexanoate-cyclic dimer as the sole source of carbon and nitrogen was newly isolated from wastewater of a factory which produces nylon-6. Two responsible enzymes, 6-aminohexanoate-cyclic-dimer hydrolase (P-EI) and 6-aminohexanoate-dimer hydrolase (P-EII), were found in the NK87 strain, as is the case with Flavobacterium sp. strain KI72, another 6-aminohexanoate-cyclic-dimer-metabolizing bacterium (H. Okada, S. Negoro, H. Kimura, and S. Nakamura, Nature [London] 306:203-206, 1983). The P-EI enzyme is immunologically identical to the 6-aminohexanoate-cyclic-dimer hydrolase of KI72 (F-EI). However, antiserum against the 6-aminohexanoate-dimer hydrolase purified from KI72 (F-EII) did not react with cell extracts of NK87, indicating that the F-EII and P-EII enzymes are immunologically different. Restriction endonuclease analyses show that the NK87 strain harbors at least six plasmids ranging in size from 20 to 80 kilobase pairs (kbp). The P-EI and P-EII genes were cloned in Escherichia coli. Both the P-EI and F-EI probes strongly hybridized with a 23-kbp plasmid in Southern hybridization analyses. The P-EII probe hybridized specifically with an 80-kbp plasmid, but the F-EII probe hybridized with none of the plasmids harbored in NK87. These results indicate that the P-EI gene and P-EII gene are encoded on the 23-kbp and 80-kbp plasmids, respectively.
Kanagawa
K
K
Department of Fermentation Technology, Osaka University, Japan.
Negoro
S
S
Takada
N
N
Okada
H
H
D000581:Amidohydrolases / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D005798:Genes, Bacterial; D009757:Nylons / Q000378:metabolism; D010957:Plasmids; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping; D015262:Xenobiotics / Q000378:metabolism
D004269:DNA, Bacterial; D009757:Nylons; D015262:Xenobiotics; D000581:Amidohydrolases; C019683:6-aminohexanoate-cyclic-dimer hydrolase