Kinetic study of enzymatic urea hydrolysis in the pH range 4-9

The enzymatic hydrolysis of urea by jack bean urease was investigated at 25degreesC over the pH range 4-9. Reaction rate data were found to be well described by a modified Michaelis-Menten equation with a pH-dependent rate coefficient and a product inhibition term. The influence of pH on activity was interpreted in terms of perturbation of the enzyme distribution among three differently protonated forms. Kinetic analysis yielded a Michaelis constant of 3.21 mmol l(-1) and indicated that the inhibition mechanism was of the fully non-competitive type, with K-P = 12.2 mmol l(-1). The estimated activation energy was 35.3 kJ mol(-1). The resulting kinetic expression was tested by comparing model predictions with the experimental behaviour observed in unbuffered media and over a long-teen period.