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Applied and Environmental Microbiology, February 2009, p. 719-728, Vol. 75, No. 3
0099-2240/09/$08.00+0     doi:10.1128/AEM.01771-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Metabolomic Investigation of the Bacterial Response to a Metal Challenge{triangledown}

Valentina Tremaroli,1,5 Matthew L. Workentine,1,2 Aalim M. Weljie,3 Hans J. Vogel,1,3 Howard Ceri,1,2 Carlo Viti,4 Enrico Tatti,4 Ping Zhang,3 Alexander P. Hynes,1 Raymond J. Turner,1* and Davide Zannoni5

Department of Biological Sciences,1 Biofilm Research Group,2 Metabolomics Research Centre, University of Calgary, Calgary, Alberta, Canada,3 Department of Agricultural Biotechnology, University of Florence, Florence, Italy,4 Department of Biology, General Microbiology Unit, University of Bologna, Bologna, Italy5

Received 31 July 2008/ Accepted 22 November 2008

Pseudomonas pseudoalcaligenes KF707 is naturally resistant to the toxic metalloid tellurite, but the mechanisms of resistance are not known. In this study we report the isolation of a KF707 mutant (T5) with hyperresistance to tellurite. In order to characterize the bacterial response and the pathways leading to tolerance, we utilized Phenotype MicroArray technology (Biolog) and a metabolomic technique based on nuclear magnetic resonance spectroscopy. The physiological states of KF707 wild-type and T5 cells exposed to tellurite were also compared in terms of viability and reduced thiol content. Our analyses showed an extensive change in metabolism upon the addition of tellurite to KF707 cultures as well as different responses when the wild-type and T5 strains were compared. Even in the absence of tellurite, T5 cells displayed a "poised" physiological status, primed for tellurite exposure and characterized by altered intracellular levels of glutathione, branched-chain amino acids, and betaine, along with increased resistance to other toxic metals and metabolic inhibitors. We conclude that hyperresistance to tellurite in P. pseudoalcaligenes KF707 is correlated with the induction of the oxidative stress response, resistance to membrane perturbation, and reconfiguration of cellular metabolism.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Calgary, 2500 University Drive NW, Calgary, Alberta T2N 1N4, Canada. Phone: (403) 220-4308. Fax: (403) 289-9211. E-mail: turnerr{at}ucalgary.ca

{triangledown} Published ahead of print on 1 December 2008.

Applied and Environmental Microbiology, February 2009, p. 719-728, Vol. 75, No. 3
0099-2240/09/$08.00+0     doi:10.1128/AEM.01771-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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