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Applied and Environmental Microbiology, May 2004, p. 3064-3072, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.3064-3072.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Growth of Polychlorinated-Biphenyl-Degrading Bacteria in the Presence of Biphenyl and Chlorobiphenyls Generates Oxidative Stress and Massive Accumulation of Inorganic Polyphosphate

Francisco P. Chávez,1 Heinrich Lünsdorf,2 and Carlos A. Jerez1*

Laboratory of Molecular Microbiology and Biotechnology and Millennium Institute for Advanced Studies in Cell Biology and Biotechnology, Department of Biology, Faculty of Sciences, University of Chile, Santiago, Chile,1 Division of Microbiology, GBF-National Research Centre for Biotechnology, Braunschweig, Germany2

Received 2 September 2003/ Accepted 22 January 2004

Inorganic polyphosphate (polyP) plays a significant role in increasing bacterial cell resistance to unfavorable environmental conditions and in regulating different biochemical processes. Using transmission electron microscopy of the polychlorinated biphenyl (PCB)-degrading bacterium Pseudomonas sp. strain B4 grown in defined medium with biphenyl as the sole carbon source, we observed large and abundant electron-dense granules at all stages of growth and following a shift from glucose to biphenyl or chlorobiphenyls. Using energy dispersive X-ray analysis and electron energy loss spectroscopy with an integrated energy-filtered transmission electron microscope, we demonstrated that these granules were mainly composed of phosphate. Using sensitive enzymatic methods to quantify cellular polyP, we confirmed that this polymer accumulates in PCB-degrading bacteria when they grow in the presence of biphenyl and chlorobiphenyls. Concomitant increases in the levels of the general stress protein GroEl and reactive oxygen species were also observed in chlorobiphenyl-grown cells, indicating that these bacteria adjust their physiology with a stress response when they are confronted with compounds that serve as carbon and energy sources and at the same time are chemical stressors.

* Corresponding author. Mailing address: Departamento de Biología, Facultad de Ciencias Universidad de Chile, Santiago 1, Casilla 653, Santiago, Chile. Phone: (56-2) 678 7376. Fax: (56-2) 271-2983. E-mail: cjerez{at}uchile.cl.

Applied and Environmental Microbiology, May 2004, p. 3064-3072, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.3064-3072.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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