Cloning and characterization of a catechol-degrading gene cluster from 3,4-dichloroaniline degrading bacterium Pseudomonas sp. KB35B.

We recently isolated a bacterium, Pseudomonas sp. KB35B, capable of growth on 3,4-dichloroaniline (DCA) as a sole carbon source. The isolated strain showed a high level of catechol 2,3-dioxygenase (CD-2,3) activity in the presence of 3,4-DCA. In an attempt to elucidate the relationship between biodegradation of 3,4-DCA and CD-2,3 activity, the genes encoding enzymes for the catabolic pathway of catechol were cloned and sequenced from the chromosomal DNA. The sequence analysis of the 10752 bp DNA fragment revealed 12 open reading frames in the order of nahRGTHINLOMKJX. Among the 12 genes, nahHINLOMK genes encode enzymes for the metabolism of catechol to TCA cycle intermediates. The nahR gene is the LysR type transcriptional regulator, and the nahH gene encodes CD-2,3 for meta-cleavages of catechol. 2-Hydroxymuconic semialdehyde hydrolase, 2-oxypent-4-dienoate hydratase, and 4-hydroxy-2-oxovalerate aldolase encoded by nahLMN genes are responsible for the three steps after meta-cleavages of catechol. The current results suggested that Pseudomonas sp. KB35B degrades 3,4-DCA via the meta-cleavage pathway of catechol.