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Microbiology 146 (2000), 2967-2975
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Microbiology (2000), 146, 2967-2975.
© 2000 Society for General Microbiology

Physiology and Growth

Growth inhibition of Escherichia coli by dichloromethane in cells expressing dichloromethane dehalogenase/glutathione S-transferase

Gareth J. Evans1, Gail P. Fergusona,1, Ian R. Booth1 and Stéphane Vuilleumier2

Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK1
Institut für Mikrobiologie, ETH Zürich, ETH-Zentrum/LFV, CH-8092 Zürich, Switzerland2

Author for correspondence: Stéphane Vuilleumier. Tel: +41 1 632 33 57. Fax: +41 1 632 11 48. e-mail: svuilleu{at}micro.biol.ethz.ch

Dichloromethane (DCM) dehalogenase converts DCM to formaldehyde via the formation of glutathione metabolites and generates 2 mol HCl per mol DCM metabolized. Growth of Escherichia coli expressing DCM dehalogenase was immediately and severely inhibited during conversion of 0·3 mM DCM. Intracellular pH (pHi) rapidly decreased and chloride ions were steadily released into the medium. Bacterial growth resumed after completion of DCM conversion and cell viability was unaffected. At 0·6 mM DCM there was no recovery from growth inhibition in liquid culture due to the build-up of inhibitory concentrations of formaldehyde. DCM turnover stimulated potassium efflux from cells, which was suppressed by glucose. The potassium efflux, therefore, did not contribute to growth inhibition. It was concluded that initial growth inhibition results from lowering of the cytoplasmic pH, but severity of growth inhibition was greater than expected for the change in pHi. Possible contributors to growth inhibition are discussed.

Keywords: glutathione, dichloromethane, intracellular pH, chloride, formaldehyde

Abbreviations: DCM, dichloromethane; FDH, formaldehyde dehydrogenase; GSH, glutathione; GST, glutathione S-transferase; pHi, intracellular pH; {Delta}{varphi}, membrane potential

a Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.






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