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Antimicrobial Agents and Chemotherapy, June 2000, p. 1556-1561, Vol. 44, No. 6
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

OXA-24, a Novel Class D beta -Lactamase with Carbapenemase Activity in an Acinetobacter baumannii Clinical Strain

Germán Bou,dagger Antonio Oliver, and Jesús Martínez-Beltrán*

Servicio de Microbiología, Hospital Ramón y Cajal, 28034 Madrid, Spain

Received 11 June 1999/Returned for modification 13 October 1999/Accepted 8 March 2000

Acinetobacter baumannii RYC 52763/97, a clinical isolate involved in a prolonged nosocomial outbreak at our hospital, was resistant to all beta -lactams tested, including imipenem and meropenem, which had MICs of 128 and 256 µg/ml, respectively. This strain synthesized three beta -lactamases: a plasmid-mediated TEM-1 beta -lactamase (pI 5.4), an AmpC-type chromosomal cephalosporinase (pI 9.4), and a novel, presumptively chromosomally mediated OXA-related enzyme (pI 9.0) named OXA-24. After cloning and sequencing, the deduced amino acid sequence of the OXA-24 beta -lactamase showed 40% homology with the OXA-10 (PSE-2) and OXA-7 beta -lactamases, 39% homology with the OXA-11 and OXA-5 enzymes, and 33% homology with the LCR-1 beta -lactamase. The amino acid sequence of the OXA-24 beta -lactamase contained the STFK motif found in serine beta -lactamases, but the typical class D triad KTG was replaced by KSG and the motif YGN was replaced by FGN. The OXA-24 beta -lactamase hydrolyzed benzylpenicillin and cephaloridine but lacked activity against oxacillin, cloxacillin, and methicillin. The enzymatic activity was inhibited by chloride ions and by tazobactam (50% inhibitory concentration [IC50], 0.5 µM), sulbactam (IC50, 40 µM), and clavulanic acid (IC50, 50 µM). Carbapenem MICs for an Escherichia coli transformant (pBMB-1) expressing the cloned OXA-24 enzyme had a fourfold increase. Relative Vmax/Km values of 13 and 6 were obtained with imipenem and meropenem, respectively, and a positive microbiological assay result with imipenem was obtained with a purified enzymatic extract of this transformant strain. Therefore, we consider this new beta -lactamase to be involved in the carbapenem resistance of A. baumannii RYC 52763/97.

* Corresponding author. Mailing address: Servicio de Microbiología, Hospital Ramón y Cajal, Carretera de Colmenar Km. 9,100, 28034 Madrid, Spain. Phone: 34-1-3368082. Fax: 34-1-3368809. E-mail: jmtzbeltran{at}hrc.insalud.es.

dagger Present address: Department of Immunology and Division of Infectious Diseases, Mayo Clinic, Rochester, MN 55905.

Antimicrobial Agents and Chemotherapy, June 2000, p. 1556-1561, Vol. 44, No. 6
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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