title: The kinetics of non-stoichiometric bursts of beta-lactam hydrolysis catalysed by class C beta-lactamases.
abstract: Class C beta-lactamases from Pseudomonas aeruginosa and several species of the Enterobacteriaceae have been observed to undergo a rapid burst in hydrolysis of beta-lactam antibiotics before relaxation to a steady-state rate of hydrolysis. The amplitude of the burst corresponds to the hydrolysis of between 1 and 10,000 mol of the substrate per mol of enzyme. The decay of the rate of hydrolysis in the burst phase comprises two exponential reactions, which indicates that there are three different reactive states of the enzymes. Examination of the kinetics of acylation by slowly reacting beta-lactams suggests that there are three forms of the free enzyme in slow equilibrium. Thus it would appear that the burst kinetics exhibited by class C enzymes can be attributed to redistribution of the enzyme between different conformations induced by the reaction with substrate.
authors.affiliation: Pharma Division, Preclinical Research, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
authors.forename: M G
authors.initials: MG
authors.lastname: Page
mesh_terms: D000215:Acylation; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D001426:Bacterial Proteins; D002513:Cephamycins / Q000378:metabolism; D004755:Enterobacteriaceae / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology; D006868:Hydrolysis; D007700:Kinetics; D008956:Models, Chemical; D010078:Oxazines / Q000378:metabolism; D010397:Penicillanic Acid / Q000031:analogs & derivatives / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013045:Species Specificity; D013997:Time Factors; D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D002513:Cephamycins; D010078:Oxazines; D011994:Recombinant Proteins; D065093:beta-Lactamase Inhibitors; D010397:Penicillanic Acid; C048795:AmpC beta-lactamases; D001618:beta-Lactamases; C037902:isatoic anhydride; C035978:cefbuperazone