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Journal of Bacteriology, June 2002, p. 3053-3060, Vol. 184, No. 11
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.11.3053-3060.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Energetics of Helicobacter pylori and Its Implications for the Mechanism of Urease-Dependent Acid Tolerance at pH 1

Kerstin Stingl,* Eva-Maria Uhlemann, Roland Schmid, Karlheinz Altendorf, and Evert P. Bakker

Abteilung Mikrobiologie, Universität Osnabrück, D-49069 Osnabrück, Germany

Received 26 October 2001/ Accepted 3 March 2002

In the presence of urea the neutrophilic human pathogen Helicobacter pylori survives for several hours at pH 1 with concomitant cytoplasmic pH homeostasis. To study this effect in detail, the transmembrane proton motive force and cytoplasmic urease activity of H. pylori were determined at various pH values. In the absence of urea, the organism maintained a close-to-neutral cytoplasm and an internally negative membrane potential at external pH values greater than 4 to 5. In the presence of urea, H. pylori accomplished cytoplasmic pH homeostasis down to an external pH of 1.2. At this external pH, the cytoplasmic pH was 4.9 and the membrane potential was slightly negative inside. The latter finding is in contrast to the situation in acidophiles, which develop inside-positive membrane potentials under similar conditions. Measurements of the time course of the membrane potential confirmed that addition of urea to the cells led to hyperpolarization. Most likely, this effect was due to electrogenic export of ammonium cations from the cytoplasm. The urease activity of intact cells increased nearly exponentially with decreasing external pH. This activation was not due to enhanced gene expression at low external pH values. In cell extracts the pH optimum of urease activity was dependent on the buffer system and was about pH 5 in sodium citrate buffer. Since this is the cytoplasmic pH of the cells at pH 1 to 2, we propose that cytoplasmic pH is a factor in the in vivo activation of the urease at low external pH values. The mechanism by which urease activity leads to cytoplasmic pH homeostasis in H. pylori is discussed.


* Corresponding author. Mailing address: Abteilung Mikrobiologie, Universität Osnabrück, D-49069 Osnabrück, Germany. Phone: 495419692867. Fax: 495419692870. E-mail: stingl{at}biologie.uni-osnabrueck.de.


Journal of Bacteriology, June 2002, p. 3053-3060, Vol. 184, No. 11
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.11.3053-3060.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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