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Antimicrobial Agents and Chemotherapy, April 1999, p. 902-906, Vol. 43, No. 4
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Biochemical Characterization of the Pseudomonas aeruginosa 101/1477 Metallo-beta -Lactamase IMP-1 Produced by Escherichia coli

Nezha Laraki,1 Nicola Franceschini,2 Gian Maria Rossolini,3 Pasqualino Santucci,2 Cécile Meunier,4 Edwin de Pauw,4 Gianfranco Amicosante,2 Jean Marie Frère,1 and Moreno Galleni1,*

Laboratoire d'Enzymologie & Centre d'Ingénierie des Protéines, Institut de Chimie,1 and Département de Chimie Générale & Chimie Physique, B6 Sart Tilman,4 Université de Liège, B-4000 Liège, Belgium, and Dipartimento di Scienze & Tecnologie Biomediche, Università dell'Aquila, I-67100 Loc Coppito, L'Aquila,2 and Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, I-53100 Siena,3 Italy

Received 1 June 1998/Returned for modification 5 October 1998/Accepted 5 January 1999

The blaIMP gene coding for the IMP-1 metallo-beta -lactamase produced by a Pseudomonas aeruginosa clinical isolate (isolate 101/1477) was overexpressed via a T7 expression system in Escherichia coli BL21(DE3), and its product was purified to homogeneity with a final yield of 35 mg/liter of culture. The structural and functional properties of the enzyme purified from E. coli were identical to those of the enzyme produced by P. aeruginosa. The IMP-1 metallo-beta -lactamase exhibits a broad-spectrum activity profile that includes activity against penicillins, cephalosporins, cephamycins, oxacephamycins, and carbapenems. Only monobactams escape its action. The enzyme activity was inhibited by metal chelators, of which 1,10-o-phenanthroline and dipicolinic acid were the most efficient. Two zinc-binding sites were found. The zinc content of the P. aeruginosa 101/1477 metallo-beta -lactamase was not pH dependent.

* Corresponding author. Mailing address: Laboratoire d'Enzymologie & Centre d'Ingénierie des Protéines, Institut de Chimie, B6 Sart Tilman, Université de Liège, B-4000 Liège, Belgium. Phone: 32-4-3663348. Fax: 32-4-3663364. E-mail: mgalleni{at}ulg.ac.be.

Antimicrobial Agents and Chemotherapy, April 1999, p. 902-906, Vol. 43, No. 4
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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