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-ketoadipate pathway genes, pcaRHGBDCFIJ, also found in proteobacteria
1 Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
2 Department of Chemistry, Faculty of Sciences, Science University of Tokyo, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162-8601, Japan
3 Department of Biotechnology, Faculty of Bioresource Sciences, Akita Prefectural University, 241-7 Kaidobata-nishi, Shimoshinjo-nakano, Akita 010-0195, Japan
4 Geo and Water Environmental Engineering Department, Obayashi Corporation, 4-640 Shimokiyoto, Kiyose-shi, Tokyo 204-0011, Japan
Correspondence
Hiroshi Habe
uhhabe{at}mail.ecc.u-tokyo.ac.jp
Terrabacter sp. strain DBF63 is capable of degrading fluorene (FN) to tricarboxylic acid cycle intermediates via phthalate and protocatechuate. Genes were identified for the protocatechuate branch of the 
-ketoadipate pathway (pcaR, pcaHGBDCFIJ) by sequence analysis of a 70 kb DNA region of the FN-catabolic linear plasmid pDBF1. RT-PCR analysis of RNA from DBF63 cells grown with FN, dibenzofuran, and protocatechuate indicated that the pcaHGBDCFIJ operon was expressed during both FN and protocatechuate degradation in strain DBF63. The gene encoding -ketoadipate enol-lactone hydrolase (pcaD) was not fused to the next gene, which encodes 
-carboxymuconolactone decarboxylase (pcaC), in strain DBF63, even though the presence of the pcaL gene (the fusion of pcaD and pcaC) within a pca gene cluster has been thought to be a Gram-positive trait. Quantitative RT-PCR analysis revealed that pcaD mRNA levels increased sharply in response to protocatechuate, and a biotransformation experiment with cis,cis-muconate using Escherichia coli carrying both catBC and pcaD indicated that PcaD exhibited -ketoadipate enol-lactone hydrolase activity. The location of the pca gene cluster on the linear plasmid, and the insertion sequences around the pca gene cluster suggest that the ecologically important -ketoadipate pathway genes, usually located chromosomally, may be spread widely among bacterial species via horizontal transfer or transposition events.
The GenBank/EMBL/DDBJ accession number for the sequence reported in this paper is AP008980.
A figure showing RT-PCR and expression analysis of pca genes, and a table of annotated ORFs are available as supplementary material with the online version of this paper.
Present address: Department of Industrial Chemistry, Faculty of Engineering, Shibaura Institute of Technology, Minato-ku, Tokyo 108-8548, Japan.
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