The THIN-B metallo-β-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.D., Lopizzo, T., Liberatori, S., Prenna, M., Thaller, M.C., Frere, 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
DOCQUIER J. D.;
2004-01-01
Abstract
The THIN-B metallo-β-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.
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https://hdl.handle.net/11365/3729
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