Kinetics and product stoichiometry of ureolysis by human salivary bacteria and artificial mouth plaques.

Ureolysis was investigated in salivary bacteria from persons with widely-differing oral ureolytic activities. Rate curves and product stoichiometry were established for urea disappearance, ammonia appearance and conversion of [14C]-urea to 14CO2. Ammonia, released stoichiometrically from urea, was best measured by a direct phenate-hypochlorite reaction. About 80 per cent of the urea-C was liberated as free CO2. Slight deviations from ammonia stoichiometry and most of the CO2 loss occurred in the first 5-10 min of reaction, when the rate of urea disappearance was constant and up to 2-fold higher than subsequently. This rate-change suggests that flux in the ureolysis pathway may be under feedback control. Ureolysis by salivary-sediment bacteria followed Michaelis-Menten kinetics with a Km of 2.5 mM; rates of end-product formation were independent of urea concentration between 25 and 500 mM. Ureolysis was inhibited 98 per cent by 5 mM acetohydroxamic acid, a urease inhibitor, and could be partly solubilized by sonication to give an enzyme preparation which, without cofactor supplementation, quantitatively hydrolysed urea. Thus urea metabolism by oral bacteria may principally involve urease-catalysed hydrolysis, rather than non-urease pathways.