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Antimicrobial Agents and Chemotherapy, July 1999, p. 1644-1650, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Biochemical-Genetic Analysis and Distribution of FAR-1, a Class A beta -Lactamase from Nocardia farcinica

Frederic Laurent,1,2 Laurent Poirel,1 Thierry Naas,1,3 El Bachir Chaibi,4 Roger Labia,4 Patrick Boiron,5 and Patrice Nordmann1,*

Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique-Hôpitaux de Paris, Faculté de Médecine Paris-Sud, 94275 Le Kremlin-Bicêtre Cédex,1 Service de Bactériologie, Hôpital Lyon-Sud, Hospices Civils de Lyon, Faculté de Médecine Lyon-Sud, 69921 Oullins Cédex,2 Service de Bactériologie-Virologie, Hôpital Antoine Béclère, Assistance Publique-Hôpitaux de Paris, Faculté de Médecine Paris-Sud, 92141 Clamart Cédex,3 UMR175 CNRS Chimie et Biologie des Substances Actives, 29000 Quimper,4 and Centre National de Référence des Mycoses, des Antifongiques et des Actinomycètes, Institut Pasteur, 75724 Paris Cédex,5 France

Received 20 July 1998/Returned for modification 9 November 1998/Accepted 23 April 1999

From genomic DNA of the clinical isolate Nocardia farcinica VIC, a 1.6-kb Sau3AI fragment was cloned and expressed in Escherichia coli JM109. The recombinant strain expressed a beta -lactamase (pI, 4.6), FAR-1, which conferred high levels of resistance to amoxicillin, piperacillin, ticarcillin, and cephalothin. The hydrolysis constants (kcat, Km, Ki, and 50% inhibitory concentration) confirmed the MIC results and showed that FAR-1 activity is inhibited by clavulanic acid and at a low level by tazobactam and sulbactam. Moreover, FAR-1 beta -lactamase hydrolyzes aztreonam (at a low level) without significant activity against ceftazidime, cefotaxime and imipenem. FAR-1 mature protein of molecular mass ca 32 kDa, has less than 60% amino acid identity with any other class A beta -lactamases, being most closely related to PEN-A from Burkholderia cepacia (52%). A blaFAR-1-like gene was found in all studied N. farcinica strains, underlining the constitutive origin of this gene.

* Corresponding author. Mailing address: Service de Bactériologie-Virologie, Hôpital de Bicêtre, 78 rue du Général Leclerc, 94275 Le Kremlin-Bicêtre Cédex, France. Phone: 33-1-45-21-36-32. Fax: 33-1-45-21-63-40. E-mail: nordmann.patrice{at}bct.ap-hop-paris.fr.

Antimicrobial Agents and Chemotherapy, July 1999, p. 1644-1650, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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