Expression of 2-halobenzoate dioxygenase genes (cbdSABC) involved in the degradation of benzoate and 2-halobenzoate in Burkholderia sp. TH2.
Burkholderia sp. TH2, isolated from soil, utilizes 2-chlorobenzoate (2CB) and benzoate (BA) as its sole source of carbon and energy. The genes for 2-halobenzoate dioxygenase (cbdABC) from Burkholderia sp. TH2 were cloned and sequenced. The predicted amino acid sequences of all the gene products are highly similar to the cbd gene products of Pseudomonas sp. 2CBS. Disruption of the promoter region of cbdA resulted in loss of growth on 2CB and BA, indicating that these genes are involved in the growth of TH2 on these substrates. Expression of the cbd genes was analyzed by transcriptional fusion assay. The cbdS gene, a possible araC/xylS-type transcriptional regulatory gene, was shown to positively regulate the expression of cbdA. In addition, the effectors of CbdS were shown to be 2CB, 2-bromobenzoate, o-toluate (2-methylbenzoate), 2-iodobenzoate, and BA. Primer extension analysis showed that the cbdA mRNA started at two positions, 14 and 15 nucleotides upstream from the cbdA start codon, ATG. A pair of direct repeats, identical to that of the Pm promoter of the TOL plasmid, was found upstream of -35 hexamer of the cbdA promoter.
Suzuki
K
K
National Institute of Agro-Environmental Sciences, 3-1-1 Kan-nondai, Tsukuba, 305-8604, Ibaraki, Japan. k1suzuki@niaes.affrc.go.jp
Ogawa
N
N
Miyashita
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D001565:Benzoates / Q000378:metabolism; D019117:Burkholderia / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D009876:Operon; D010105:Oxygenases / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D001565:Benzoates; D010105:Oxygenases; C072183:2-chlorobenzoate 1,2-dioxygenase