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J Antimicrob Chemother. 2015 Apr;70(4):1059-63. doi: 10.1093/jac/dku524. Epub 2015 Jan 11.

Heterogeneous hydrolytic features for OXA-48-like β-lactamases.

Author information

1
INSERM U914 'Emerging Resistance to Antibiotics', K.-Bicêtre, France LabEx LERMIT, Faculté de Médecine Paris Sud, K.-Bicêtre, France.
2
INSERM U914 'Emerging Resistance to Antibiotics', K.-Bicêtre, France LabEx LERMIT, Faculté de Médecine Paris Sud, K.-Bicêtre, France Centre National Associé-Centre de Référence des Résistances aux Antibiotiques, K.-Bicêtre, France Medical and Molecular Microbiology, Department of Medicine, Faculty of Science, University of Fribourg, Fribourg, Switzerland HFR-Hôpital Cantonal, Fribourg, Switzerland.
3
INSERM U914 'Emerging Resistance to Antibiotics', K.-Bicêtre, France LabEx LERMIT, Faculté de Médecine Paris Sud, K.-Bicêtre, France Centre National Associé-Centre de Référence des Résistances aux Antibiotiques, K.-Bicêtre, France Medical and Molecular Microbiology, Department of Medicine, Faculty of Science, University of Fribourg, Fribourg, Switzerland laurent.poirel@unifr.ch.

Abstract

OBJECTIVES:

Carbapenem-hydrolysing class D β-lactamases of the OXA-48 type are increasingly reported from Enterobacteriaceae. β-Lactamase OXA-48 hydrolyses penicillins very efficiently, but carbapenems only weakly and spares broad-spectrum cephalosporins. Recently, diverse OXA-48-like β-lactamases have been identified worldwide (OXA-162, OXA-181, OXA-163, OXA-204 and OXA-232). They differ by few amino acid substitutions or by amino acid deletions.

METHODS:

blaOXA-48, blaOXA-162, blaOXA-163, blaOXA-181, blaOXA-204 and blaOXA-232 were cloned into the same expression vector and expressed in the same Escherichia coli background. Kinetic studies were performed with enzymes purified by ion-exchange chromatography. Determination of hydrolytic activities was performed by UV spectrophotometry. MICs were determined for all recombinant strains, using as background either the WT E. coli TOP10 strain or a porin-deficient E. coli strain.

RESULTS:

Kinetic studies showed that OXA-162 and OXA-204 shared the same hydrolytic properties as OXA-48. On the other hand, OXA-181 possessed a higher ability to hydrolyse carbapenems, while OXA-232 hydrolysed those substrates less efficiently. In contrast to the other OXA-48-like β-lactamases, OXA-163 hydrolysed broad-spectrum cephalosporins very efficiently, but did not possess significant carbapenemase activity. Although several of these OXA-48-like enzymes possess low activity against carbapenems, MICs of carbapenems were significantly elevated when determined for strains possessing permeability defects.

CONCLUSIONS:

A detailed comparative analysis of the kinetic properties of the OXA-48-like β-lactamases is provided here. It clarifies the respective features of each OXA-48-like variant and their respective impacts in terms of carbapenem resistance.

KEYWORDS:

carbapenemases; class D β-lactamases; enzymatic activity

PMID:
25583748
DOI:
10.1093/jac/dku524
[Indexed for MEDLINE]

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