The urease-catalyzed hydrolysis of thiourea and thioacetamide

Jack bean urease catalyzes the hydrolysis of thiourea with a second-order rate constant (K-cat/K-m) of 1.6 (+/-0.2) x 10(-3) M-1 s(-1) at pH7, 25 degrees C. This value is lower than that for urea by a factor of 3 x 10(8). The corresponding substitution of S for O in acetamide reduces the k(cat)/K-m value by only a factor of 33. This greater reactivity of the oxo compounds than of the corresponding thiono compounds, and the tighter binding of urea (K-s = 2.9 mM) than of either the guanidinium ion (K-i = 30 mM) or thiourea (K-i = 70 mM), suggests that the substrate chalcogen (S or O) is more likely to be stabilized in the transition state by coordination to the enzyme via a neutral hydrogen-bond donor (i.e., Bronsted acid catalysis) than by coordination via one of the active-site nickel ions (i.e., Lewis acid catalysis). (C) 1998 Academic Press.