The Legionella (Fluoribacter) gormanii metallo-β-lactamase: A new member of the highly divergent lineage of molecular-subclass B3 β-lactamases

Letizia Boschi, Paola Sandra Mercuri, Maria Letizia Riccio, Gianfranco Amicosante, Moreno Galleni, Jean Marie Frère, Gian Maria Rossolini

Research output: Contribution to journalArticle

Abstract

A metallo-β-lactamase determinant was cloned from a genomic library of Legionella (Fluoribacter) gormanii ATCC 33297(T) constructed in the plasmid vector pACYC184 and transformed into Escherichia coli DH5α, by screening for clones showing a reduced susceptibility to imipenem. The product of the cloned determinant, named FEZ-1, contains a 30-kDa polypeptide and exhibits an isoelectric pH of 7.6. Sequencing revealed that FEZ-1 is a molecular-class B β-lactamase which shares the closest structural similarity (29.7% of identical residues) with the L1 enzyme of Stenotrophomonas maltophilia, being a new member of the highly divergent subclass B3 lineage. All the residues that in L1 are known to be directly or indirectly involved in coordination of the zinc ions were found to be conserved also in FEZ-1, suggesting that the geometry of zinc coordination in the active site of the latter enzyme is identical to that of L1. Unlike L1, however, FEZ-1 appeared to be monomeric in gel permeation chromatography experiments and exhibited a distinctive substrate specificity with a marked preference for cephalosporins and meropenem. The properties of FEZ-1 overall resembled those of a β-lactamase previously purified from the same strain of L. gormanii (T. Fujii, K. Sato, K. Miyata, M. Inoue, and S. Mitsuhashi, Antimicrob. Agents Chemother. 29:925- 926, 1986) and are as yet unique among class B enzymes, reinforcing the notion that considerable functional heterogeneity can be encountered among members of this class. A system for overexpression of the bla(FEZ-1) gene in E. coli, based on the T7 phage promoter, was also developed.

Original languageEnglish
Pages (from-to)1538-1543
Number of pages6
JournalAntimicrobial Agents and Chemotherapy
Volume44
Issue number6
DOIs
Publication statusPublished - 2000

Fingerprint

Legionella
meropenem
Zinc
Enzymes
Escherichia coli
Stenotrophomonas maltophilia
Bacteriophage T7
Genomic Library
Imipenem
Cephalosporins
Substrate Specificity
Gel Chromatography
Catalytic Domain
Plasmids
Clone Cells
Ions
Peptides
Genes

ASJC Scopus subject areas

  • Pharmacology (medical)

Cite this

The Legionella (Fluoribacter) gormanii metallo-β-lactamase : A new member of the highly divergent lineage of molecular-subclass B3 β-lactamases. / Boschi, Letizia; Mercuri, Paola Sandra; Riccio, Maria Letizia; Amicosante, Gianfranco; Galleni, Moreno; Frère, Jean Marie; Rossolini, Gian Maria.

In: Antimicrobial Agents and Chemotherapy, Vol. 44, No. 6, 2000, p. 1538-1543.

Research output: Contribution to journalArticle

Boschi, Letizia ; Mercuri, Paola Sandra ; Riccio, Maria Letizia ; Amicosante, Gianfranco ; Galleni, Moreno ; Frère, Jean Marie ; Rossolini, Gian Maria. / The Legionella (Fluoribacter) gormanii metallo-β-lactamase : A new member of the highly divergent lineage of molecular-subclass B3 β-lactamases. In: Antimicrobial Agents and Chemotherapy. 2000 ; Vol. 44, No. 6. pp. 1538-1543.
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