Imipenem as substrate and inhibitor of β-lactamases

Abstract

The interaction between imipenem, a carbapenem antibiotic, and two representative β-lactamases has been studied. The first enzyme was β-lactamase I, a class-A β-lactamase from Bacillus cereus; imipenem behaved as a slow substrate (k(cat.) 6.7 min-1, K(m) 0.4 mM at 30°C and at pH 7) that reacted by a branched pathway. There was transient formation of an altered species formed in a reversible reaction; this species was probably an acyl-enzyme in a slightly altered, but considerably more labile, conformation. The kinetics of the reaction were investigated by measuring both the concentration of the substrate and the activity of the enzyme, which fell and then rose again more slowly. The second enzyme was the chromosomal class-C β-lactamase from Pseudomonas aeruginosa; imipenem was a substrate with a low k(cat.) (0.8 min-1) and a low K(m) (0.7 μM). Possible implications for the clinical use of imipenem are considered.