作者: Jed Fisher , Robert L. Charnas , Jeremy R. Knowles
DOI: 10.1021/BI00604A024
关键词: Hydroxylamine 、 Enzyme assay 、 Clavulanic acid 、 Isoelectric focusing 、 Biochemistry 、 Escherichia coli 、 Absorbance 、 Chemistry 、 Enzyme 、 Polyacrylamide
摘要: Incubation of clavulanic acid with the beta-lactamase from Escherichia coli RTEM leads to enzyme-catalyzed depletion acid, transient inhibition, and irreversible inactivation enzyme. Both transiently inhibited irreversibly inactivated species show a marked increase in absorbance at 281 nm that is proportional decrease enzyme activity. Hydroxylamine treatment restores about one-third catalytic activity, concomitant nm. Polyacrylamide isoelectric focusing shows three bands approximately equal intensity, different native Upon hydroxylamine treatment, one disappears now focuses identically It evident by an excess generates products, which can be reactivated hydroxylamine.