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J. Bacteriol. doi:10.1128/JB.01675-06
Copyright (c) 2006, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Biochemical and Genetic Analysis of the -Resorcylate (2,6-Dihydroxybenzoate), Catabolic Pathway in Rhizobium sp. Strain MTP-10005: Identification and Functional Analysis of Its Gene Cluster

Department of Biotechnology, Faculty of Engineering, Kansai University, 3-3-35 Yamate-Cho, Suita, Osaka-Fu 564-8680, Kansai University High technology Research Center, Suita, Osaka-Fu 564-8680, Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto-Fu 611-0011, Japan

^* To whom correspondence should be addressed. Email: oikawa{at}ipcku.kansai-u.ac.jp,

	Abstract

We identified a gene cluster that is involved in the -resorcylate (2,6-dihydroxybenzoate) catabolism of the aerobic bacterium, Rhizobium sp. strain MTP-10005. The cluster consists of the graRDAFCBEK genes, and graA, graB, graC, and graD were heterologously expressed in Escherichia coli. The enzymological studies showed that graD, graA, graC, and graB encode the reductase (GraD) and oxygenase (GraA) components of a resorcinol hydroxylase (EC 1.14.13.x), a maleylacetate reductase (GraC; EC 1.3.1.32), and a hydroxyquinol 1,2-dioxygenase (GraB; EC 1.13.11.37). The bioinformatic analyses suggested that graE, graR, and graK encode a protein with an unknown function (GraE), a MarR-type transcriptional regulator (GraR), and a benzoate transporter (GraK). The quantitative reverse-transcription PCR of graF, which encodes -resorcylate decarboxylase, revealed that the maximum relative mRNA expression level ( (5.93±0.82) x 10^-4) of graF was detected in the total RNA of the cells after one hour cultivation when -resorcylate was used as a sole carbon source. The reverse-transcription PCR of graDAFCBE genes showed that these genes are transcribed as a single mRNA and the transcription of the gene cluster is induced by -resorcylate. These results suggested that graDAFCBE genes are responsible as an operon for the growth of Rhizobium sp. strain MTP-10005 on -resorcylate and are probably regulated by GraR at the transcription level. This is the first report of the -resorcylate catabolic pathway in an aerobic bacterium.

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