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Journal of Bacteriology, September 2001, p. 5285-5292, Vol. 183, No. 18
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.18.5285-5292.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Involvement of the TonB System in Tolerance to Solvents and Drugs in Pseudomonas putida DOT-T1E

Patricia Godoy, María Isabel Ramos-González, and Juan L. Ramos*

Department of Biochemistry and Molecular and Cellular Biology of Plants, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, E-18008 Granada, Spain

Received 29 May 2001/Accepted 3 July 2001

Pseudomonas putida DOT-T1E is able to grow with glucose as the carbon source in liquid medium with 1% (vol/vol) toluene or 17 g of (123 mM) p-hydroxybenzoate (4HBA) per liter. After random mini-Tn5'phoA-Km mutagenesis, we isolated the mutant DOT-T1E-PhoA5, which was more sensitive than the wild type to 4HBA (growth was prevented at 6 g/liter) and toluene (the mutant did not withstand sudden toluene shock). Susceptibility to toluene and 4HBA resulted from the reduced efflux of these compounds from the cell, as revealed by accumulation assays with 14C-labeled substrates. The mutant was also more susceptible to a number of antibiotics, and its growth in iron-deficient minimal medium was inhibited in the presence of ethylenediamine-di(o-hydroxyphenylacetic acid (EDDHA). Cloning the mutation in the PhoA5 strain and sequencing the region adjacent showed that the mini-Tn5 transposor interrupted the exbD gene, which forms part of the exbBD tonB operon. Complementation by the exbBD and tonB genes cloned in pJB3-Tc restored the wild-type characteristics to the PhoA5 strain.

* Corresponding author. Mailing address: Estación Experimental del Zaidín, C/Prof. Albareda 1, E-18008 Granada, Spain. Phone: 34 958 121011. Fax: 34 958 129600. E-mail: jlramos{at}eez.csic.es.

Journal of Bacteriology, September 2001, p. 5285-5292, Vol. 183, No. 18
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.18.5285-5292.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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  • Huang, B., Ru, K., Yuan, Z., Whitchurch, C. B., Mattick, J. S. (2004). tonB3 Is Required for Normal Twitching Motility and Extracellular Assembly of Type IV Pili. J. Bacteriol. 186: 4387-4389 [Abstract] [Full Text]  
  • Ramos-Gonzalez, M.-I., Ben-Bassat, A., Campos, M.-J., Ramos, J. L. (2003). Genetic Engineering of a Highly Solvent-Tolerant Pseudomonas putida Strain for Biotransformation of Toluene to p-Hydroxybenzoate. Appl. Environ. Microbiol. 69: 5120-5127 [Abstract] [Full Text]  


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