Urease activity in Streptococcus salivarius at low pH

Abstract

Arginine metabolism to alkali by the arginine deiminase system in oral bacteria increases their acid tolerance. The potential of urease activity in Streptococcus salivarius to fulfil a similar role was examined. In cell extracts between pH 5.0 and 8.0, urease activity was over 80% the maximal rate. The urease rate was zero at pH 4.3, and at pH 3.6 the enzyme was rapidly inactivated ( t 1 2 of 0.6 min ). The pH range of intact cells was broader. In Strep. salivarius cells acidified to pH 2.6 for 5 min, urease was completely retained and the ureolytic pH rise was rapid. There was no urease activity after acidification to pH 2. In cells acidified to maintain the pH between 3.3 and 4, viability was maintained for a short period (extrapolation indicated 20 min) and then decreased. This acidification induced alkali generation or acid removal that decreased in parallel to loss of viability. A small fraction (10%) of the urease was rapidly inactivated, after which both the remaining urease and pH response decreased at a similar rate to cell viability ( t 1 2 of 15–20 min ), but for at least 1 h following acidification, a rapid ureolysis induced rise in pH to above 7. In cells held at pH 3.6 and treated to compromise their membranes by freeze-thawing or transient acidification to pH 2.3, 70–80% of the urease was lost rapidly and the remainder inactivated at a rate similar to that in intact cells. Therefore, although at pH below 4, S. salivarius urease is outside its pH activity range and the free enzyme is rapidly inactivated, intact cells the urease is protected and ureolytic generation of ammonia is capable of substantially raising the pH for at least 1 h while the cell population is being progressively killed by acid.