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Journal of Bacteriology, June 2006, p. 3862-3869, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.01675-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Metabolism of Isovanillate, Vanillate, and Veratrate by Comamonas testosteroni Strain BR6020{dagger}

Miguel A. Providenti,1,2* Jason M. O'Brien,1 Jürgen Ruff,2 Alasdair M. Cook,2 and Iain B. Lambert1

Institute of Biology, Carleton University, Ottawa, Ontario, Canada K1S 5B6,1 Department of Biology, The University of Konstanz, D-78457 Konstanz, Germany2

Received 3 November 2005/ Accepted 23 March 2006

In Comamonas testosteroni strain BR6020, metabolism of isovanillate (iVan; 3-hydroxy-4-methoxybenzoate), vanillate (Van; 4-hydroxy-3-methoxybenzoate), and veratrate (Ver; 3,4-dimethoxybenzoate) proceeds via protocatechuate (Pca; 3,4-dihydroxybenzoate). A 13.4-kb locus coding for the catabolic enzymes that channel the three substrates to Pca was cloned. O demethylation is mediated by the phthalate family oxygenases IvaA (converts iVan to Pca and Ver to Van) and VanA (converts Van to Pca and Ver to iVan). Reducing equivalents from NAD(P)H are transferred to the oxygenases by the class IA oxidoreductase IvaB. Studies using whole cells, cell extracts, and reverse transcriptase PCR showed that degradative activity and expression of vanA, ivaA, and ivaB are inducible. In succinate- and Pca-grown cells, there is negligible degradative activity towards Van, Ver, and iVan and little to no expression of vanA, ivaA, and ivaB. Growth on Van or Ver results in production of oxygenases with activity towards Van, Ver, and iVan and expression of vanA, ivaA, and ivaB. With iVan-grown cultures, ivaA and ivaB are expressed, and in assays with whole cells, production of the iVan oxygenase is observed, but there is little activity towards Van or Ver. In cell extracts, though, Ver metabolism is observed, which suggests that the system mediating iVan uptake in whole cells does not mediate Ver uptake.


* Corresponding author. Mailing address: Institute of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, Ontario, Canada K1S 5B6. Phone: (613) 520-2600, ext. 1216. Fax: (613) 520-3539. E-mail: miguel_providenti{at}carleton.ca.

{dagger} Dedicated to the memory of R. Campbell Wyndham (1951-2002).


Journal of Bacteriology, June 2006, p. 3862-3869, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.01675-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.







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