1. The rate of hydrolysis of methicillin, cloxacillin and quinacillin by staphylococcal extracellular penicillinase decreases progressively with time. 2. The inactivation is prevented but not reversed by benzylpenicillin. 3. The rate of inactivation produced by quinacillin is minimal when the rate of hydrolysis is at a maximum. 4. Under certain conditions, partially inactivated enzyme can be reactivated. 5. Combination of the enzyme with antiserum, while permitting hydrolysis, prevents inactivation. 6. No evidence for a stable enzyme-substrate complex has been found.
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