[Cloning and characterization of a chromosome-encoded catechol 2,3-dioxygenase gene from Pseudomonas aeruginosa ZD 4-3].
Catechol 2,3-dioxygenase (C23O), one of extradiol-type dioxygenases cleaving the aromatic C-C bond at the meta-position of dihydroxylated aromatic substrates, catalyzes the conversion of catechol to 2-hydroxymuconic semialdehyde. Based on curing experiment, PCR identification, and Southern hybridization, the gene responsible for C23O was localized on a 3.5-kb EcoRI/BamHI fragment and cloned from P. aeruginosa ZD 4-3 able to degrade both single and bicyclic compounds via the meta-cleavage pathway. A complete nucleotide sequence analysis of the C23O revealed that it had one ORF, which showed a strong amino acid sequence similarity to the known C23Os of mesophilic gram-negative bacteria. The alignment analysis indicated that distinct difference existed between the C23O in this study and the 2,3-dihydroxybiphenyl dioxygenases cleaving bicyclic aromatic compounds. The heterogenous expression of the pheB gene in Escherichia coli BL21(DE3) demonstrated that this C23O possessed a meta-cleavage activity.
Chen
Y -X
Y-
Liu
H
H
Zhu
L -C
L-
Jin
Y -F
Y-
D000595:Amino Acid Sequence; D050557:Catechol 2,3-Dioxygenase; D002396:Catechols / Q000378:metabolism; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D049308:Dioxygenases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010802:Phylogeny; D011550:Pseudomonas aeruginosa / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D016415:Sequence Alignment
D002396:Catechols; D011994:Recombinant Proteins; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase; C074272:2,3-dihydroxybiphenyl oxygenase; C034221:catechol