The THIN-B metallo-beta-lactamase, a subclass B3 enzyme produced by the environmental species Janthinobacterium lividum, was overproduced in Escherichia coli by means of a T7-based expression system. The enzyme was purified (>95%) by two ion-exchange chromatography steps and subjected to biochemical analysis. The native THIN-B enzyme is a monomeric protein of 31 kDa. It exhibits the highest catalytic efficiencies with carbapenem substrates and cephalosporins, except for cephaloridine, which acts as a poor inactivator. Individual rate constants for inactivation by chelators were measured, suggesting that inactivation occurred by a mechanism involving formation of a ternary complex.

Docquier, J., Lopizzo, T., Liberatori, S., Prenna, M., Thaller, M.c., Frère, J., et al. (2004). Biochemical characterization of the THIN-B metallo-beta-lactamase of Janthinobacterium lividum. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 48(12), 4778-4783 [10.1128/AAC.48.12.4778-4783.2004].

Biochemical characterization of the THIN-B metallo-beta-lactamase of Janthinobacterium lividum

THALLER, MARIA CRISTINA;
2004-12-01

Abstract

The THIN-B metallo-beta-lactamase, a subclass B3 enzyme produced by the environmental species Janthinobacterium lividum, was overproduced in Escherichia coli by means of a T7-based expression system. The enzyme was purified (>95%) by two ion-exchange chromatography steps and subjected to biochemical analysis. The native THIN-B enzyme is a monomeric protein of 31 kDa. It exhibits the highest catalytic efficiencies with carbapenem substrates and cephalosporins, except for cephaloridine, which acts as a poor inactivator. Individual rate constants for inactivation by chelators were measured, suggesting that inactivation occurred by a mechanism involving formation of a ternary complex.
dic-2004
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore MED/07 - MICROBIOLOGIA E MICROBIOLOGIA CLINICA
Settore BIO/19 - MICROBIOLOGIA GENERALE
English
Con Impact Factor ISI
beta-Lactamases; Escherichia coli; Electrophoresis, Polyacrylamide Gel; Chelating Agents; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Cephalosporins; Culture Media; Kinetics; Enzyme Inhibitors; Recombinant Proteins; Sulfhydryl Compounds; Molecular Sequence Data; Trypsin; Proteobacteria; Amino Acid Sequence; Chromatography, Ion Exchange
Docquier, J., Lopizzo, T., Liberatori, S., Prenna, M., Thaller, M.c., Frère, J., et al. (2004). Biochemical characterization of the THIN-B metallo-beta-lactamase of Janthinobacterium lividum. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 48(12), 4778-4783 [10.1128/AAC.48.12.4778-4783.2004].
Docquier, J; Lopizzo, T; Liberatori, S; Prenna, M; Thaller, Mc; Frère, J; Rossolini, G
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/9737
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