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Applied and Environmental Microbiology, May 2007, p. 2769-2776, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02584-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Genetic Characterization of 2,4,6-Trichlorophenol Degradation in Cupriavidus necator JMP134{triangledown}

M. A. Sánchez and B. González*

Laboratorio de Microbiología, Departamento de Genética Molecular y Microbiología, Millennium Nucleus on Microbial Ecology and Environmental Microbiology and Biotechnology, and Center for Advanced Studies in Ecology and Biodiversity, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile

Received 6 November 2006/ Accepted 13 February 2007

The degradation pathway of 2,4,6-trichlorophenol (2,4,6-TCP), a hazardous pollutant, in the aerobic bacterium Cupriavidus necator JMP134(pJP4) (formerly Ralstonia eutropha JMP134) is encoded by the tcp genes. These genes are located in a genetic context, tcpRXABCYD, which resembles a putative catabolic operon. In this work, these gene sequences were individually disrupted and mutant strains were evaluated for their ability to grow on or degrade 2,4,6-TCP. The tcpX and tcpA mutants completely failed to degrade this compound. Although the tcpC mutant was also unable to grow on 2,4,6-TCP, it still transformed this chlorophenol to 6-chlorohydroquinol. In contrast, the tcpD mutant grew on 2,4,6-TCP, suggesting the presence of additional maleylacetate reductase-encoding genes. Five other open reading frames encoding maleylacetate reductases, in addition to the tcpD gene, were found in the genome of C. necator, and two of them provide this function in the tcpD mutant. The tcpR gene, encoding a putative LysR-type transcriptional regulator, was disrupted, and this mutant strain completely failed to grow on 2,4,6-TCP. Transcriptional fusion studies demonstrated that TcpR activates the expression of the tcp genes, responding specifically to 2,4,6-TCP. The transcriptional start of the tcp operon was mapped, and a putative {sigma}70-type promoter was identified.

* Corresponding author. Mailing address: Laboratorio de Microbiología, Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Casilla 114-D, Santiago, Chile. Phone: 56-2-6862845. Fax: 56-2-6862185. E-mail: bgonzalez{at}bio.puc.cl

{triangledown} Published ahead of print on 23 February 2007.

Applied and Environmental Microbiology, May 2007, p. 2769-2776, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02584-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Perez-Pantoja, D., Donoso, R. A., Sanchez, M. A., Gonzalez, B. (2009). Genuine genetic redundancy in maleylacetate-reductase-encoding genes involved in degradation of haloaromatic compounds by Cupriavidus necator JMP134. Microbiology 155: 3641-3651 [Abstract] [Full Text]  
  • Belchik, S. M., Xun, L. (2008). Functions of Flavin Reductase and Quinone Reductase in 2,4,6-Trichlorophenol Degradation by Cupriavidus necator JMP134. J. Bacteriol. 190: 1615-1619 [Abstract] [Full Text]  


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