Helicobacter pylori requires an acidic environment to survive in the presence of urea

The aim of this work was to study the significance of the urease enzyme in promoting Helicobacter pylori survival in various environments. A urease-positive H. pylori isolate, strain N6, and an isogenic urease-negative strain, strain N6(ureB::TnKm), were incubated in phosphate-buffered saline at a pH ranging from 2.2 to 7.2 for 60 min at 37 degrees C in both the presence and the absence of 10 mM urea. The number of CFU per milliliter in each solution, the pH of the bacterial supernatant, and the amounts of ammonia present in the solutions were measured. H. pylori N6 survived well in solutions with pH values ranging from 4.5 to 7.0 in the absence of urea but survived in solutions only with an initial pH below 3.5 in the presence of urea. Neither strain grew after incubation in an alkaline environment. The pH of an acidic solution (i.e., 3.5) rose rapidly to 8.45 in the presence of the wild-type strain and urea. The urease-negative mutant survived in solutions with pH values ranging from 4.5 to 7.2 irrespective of the presence of urea. Ammonia was present in significant amounts when H. pylori N6 was incubated in the presence of urea. Strain N6 survived exposure to concentrations of ammonia as high as 80 mM. The acid environment of the stomach may be crucial for H. pylori survival in the presence of urea. H. pylori does not survive in the normal environment in the presence of urea because of the subsequent rise in pH rather than ammonia toxicity.