The effect of inhibitors of β-lactamases on β-lactamase extracts and on intact cells

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Abstract
The activity of β-lactamase inhibitors was measured on β-lactamases extracted from Gram-negative rods. Ampicillin and cephaloridine were used as substrates in a biological and spectrophotometric system respectively. The effect of the β-lactamases was also assessed in living bacterial cells in order to assess the ability of the inhibitors to inhibit the enzymes inside the cell. Variation in activity in inhibition was present as judged by ability to protect the two substrates from hydrolysis, sulbactam for example showing little inhibitory effect against K1 and SHV 1 enzymes whereas the enzyme was inhibited by clavulanic acid and UK 38006. Clavulanic acid is able to penetrate Pseudomonas aeruginosa, Citrobacter and Enterobacter strains, as judged by the marked fall in MIC of carbenicillin and furbenicillin in the presence of clavulanic acid. Other inhibitors were more variable in their ability to affect growing cells.

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