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.
-Lactamase IMP-1 Produced by
Escherichia coli
Laboratoire d'Enzymologie & Centre d'Ingénierie des
Protéines,
Received 1 June 1998/Returned for modification 5 October
1998/Accepted 5 January 1999
The blaIMP gene coding for the IMP-1
metallo-
-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-
-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-
-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.
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