Antibiotic Trapping by Plasmid-Encoded CMY-2 beta-Lactamase Combined with Reduced Outer Membrane Permeability as a Mechanism of Carbapenem Resistance in Escherichia coli

W.H.F. Goessens, A.K. van der Bij, R. van Boxtel, J.D.D. Pitout, J.P. van Ulsen, D.C. Melles, J. Tommassen

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

A liver transplant patient was admitted with cholangitis, for which meropenem therapy was started. Initial cultures showed a carbapenem-susceptible (CS) Escherichia coli strain, but during admission, a carbapenem-resistant (CR) E. coli strain was isolated. Analysis of the outer membrane protein profiles showed that both CS and CR E. coli lacked the porins OmpF and OmpC. Furthermore, PCR and sequence analysis revealed that both CS and CR E. coli possessed blaCTX-M-15 and blaOXA-1. The CR E. coli strain additionally harbored blaCMY-2 and demonstrated a>15-fold increase in-lactamase activity against nitrocefin, but no hydrolysis of meropenem was detected. However, nitrocefin hydrolysis appeared strongly inhibited by meropenem. Furthermore, the CMY-2 enzyme demonstrated lower electrophoretic mobility after its incubation either in vitro or in vivo with meropenem, indicative of its covalent modification with meropenem. The presence of the acyl-enzyme complex was confirmed by mass spectrometry. By transformation of the CMY-2-encoding plasmid into various E. coli strains, it was established that both porin deficiency and high-level expression of the enzyme were needed to confer meropenem resistance. In conclusion, carbapenem resistance emerged by a combination of elevated-lactamase production and lack of porin expression. Due to the reduced outer membrane permeability, only small amounts of meropenem can enter the periplasm, where they are trapped but not degraded by the large amount of the-lactamase. This study, therefore, provides evidence that the mechanism of trapping by CMY-2-lactamase plays a role in carbapenem resistance. Copyright © 2013, American Society for Microbiology. All Rights Reserved.
Original languageEnglish
Pages (from-to)3941-3949
JournalAntimicrobial Agents and Chemotherapy
Volume57
Issue number8
DOIs
Publication statusPublished - 2013

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