The bla(FEZ-1) gene coding for the metallo-beta -lactamase of Legionella (Fluoribacter) gormanii ATCC 33297(T) was overexpressed via a T7 expression system in Escherichia coli BL21(DE3)(pLysS), The product was purified to homogeneity in two steps with a yield of 53%. The FEZ-1 metallo-beta -lactamase exhibited a broad-spectrum activity profile, with a preference for cephalosporins such as cephalothin, cefuroxime, and cefotaxime, Monobactams were not hydrolyzed, The beta -lactamase was inhibited by metal chelators, FEZ-1 is a monomeric enzyme with a molecular mass of 29,440 Da which possesses two zinc-binding sites. Its zinc content did not vary in the pH range of 5 to 9, but the presence of zinc ions modified the catalytic efficiency of the enzyme. A model of the FEZ-1 three-dimensional structure was built.
Mercuri, P.S., Bouillenne, F., Boschi, L., Lamotte-Brasseur, J., Amicosante, G., Devreese, B., et al. (2001). Biochemical characterization of the FEZ-1 metallo-beta-lactamase of Legionella gormanii ATCC 33297T produced in Escherichia coli. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 45(4), 1254-1262 [10.1128/AAC.45.4.1254-1262.2001].
Biochemical characterization of the FEZ-1 metallo-beta-lactamase of Legionella gormanii ATCC 33297T produced in Escherichia coli
Rossolini G. M.;
2001-01-01
Abstract
The bla(FEZ-1) gene coding for the metallo-beta -lactamase of Legionella (Fluoribacter) gormanii ATCC 33297(T) was overexpressed via a T7 expression system in Escherichia coli BL21(DE3)(pLysS), The product was purified to homogeneity in two steps with a yield of 53%. The FEZ-1 metallo-beta -lactamase exhibited a broad-spectrum activity profile, with a preference for cephalosporins such as cephalothin, cefuroxime, and cefotaxime, Monobactams were not hydrolyzed, The beta -lactamase was inhibited by metal chelators, FEZ-1 is a monomeric enzyme with a molecular mass of 29,440 Da which possesses two zinc-binding sites. Its zinc content did not vary in the pH range of 5 to 9, but the presence of zinc ions modified the catalytic efficiency of the enzyme. A model of the FEZ-1 three-dimensional structure was built.
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https://hdl.handle.net/11365/27469
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