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Microbiology 148 (2002), 2215-2222
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Microbiology (2002), 148, 2215-2222.
© 2002 Society for General Microbiology

Research Paper

Inhibition of Escherichia coli growth by acetic acid: a problem with methionine biosynthesis and homocysteine toxicity

Andrew J. Roea,1, Conor O’Byrneb,1, Debra McLaggan1 and Ian R. Booth1

Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK1

Author for correspondence: Ian R. Booth. Tel: +44 1224 273152. Fax: +44 1224 273144. e-mail: gen118{at}abdn.ac.uk

The mechanism by which methionine relieves the growth inhibition of Escherichia coli K-12 that is caused by organic weak acid food preservatives was investigated. In the presence of 8 mM acetate the specific growth rate of E. coli Frag1 (in MacIlvaine’s minimal medium pH 6·0) is reduced by 50%. Addition of methionine restores growth to 80% of that observed in untreated controls. Similar relief was seen with cultures treated with either benzoate or propionate. Mutants with an elevated intracellular methionine pool were almost completely resistant to the inhibitory effects of acetate, suggesting that the methionine pool becomes limiting for growth in acetate-treated cells. Measurement of the intracellular concentrations of pathway intermediates revealed that the homocysteine pool is increased dramatically in acetate-treated cells, suggesting that acetate inhibits a biosynthetic step downstream from this intermediate. Supplementation of the medium with homocysteine inhibits the growth of E. coli cells. Acetate inhibition of growth arises from the depletion of the intracellular methionine pool with the concomitant accumulation of the toxic intermediate homocysteine and this augments the effect of lowering cytoplasmic pH.

Keywords: enteric bacteria, amino acid synthesis, acetate, weak acids, metabolite toxicity

Abbreviations: pHi, intracellular pH; pHo, external pH

a Present address: Zoonotic and Animal Pathogens Research Group, Department of Veterinary Pathology, Teviot Place, Edinburgh EH8 9AG, UK.

b Present address: Department of Microbiology, National University of Ireland – Galway, Galway, Ireland.




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