Overproduction and biochemical characterization of the Chryseobacterium meningosepticum B1aB metallo-β-lactamase

Sandrine Vessillier, Jean Denis Docquier, Sandrine Rival, Jean Marie Frere, Moreno Galleni, Gianfranco Amicosante, Gian Maria Rossolini, Nicola Franceschini

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Abstract

The BlaB metallo-β-lactamase of Chryseobacterium meningosepticum CCUG4310 was overproduced in Escherichia coli by means of a T7 promoter-based expression system. The overproducing system, scaled up in a 15-liter fermentor, yielded approximately 10 mg of BlaB protein per liter, mostly released in the culture supernatant. The enzyme was purified by two ion-exchange chromatographic steps with an overall yield of 66%. Analysis of the kinetic parameters revealed efficient activities (kcat/Km ratios of > 106 M-1 s-1 toward most penam and carbapenem compounds, with the exception of the 6- α-methoxypenam derivative temocillin and of biapenem, which were poorer substrates. Hydrolysis of cephalosporins was overall less efficient, with a remarkable variability that was largely due to variable affinities of the BlaB enzyme for different compounds. BlaB was also able to hydrolyze serine-β-lactamase inhibitors, including β-iodopenicillanate, sulbactam and, although less efficiently, tazobactam.

Original languageEnglish
Pages (from-to)1921-1927
Number of pages7
JournalAntimicrobial Agents and Chemotherapy
Volume46
Issue number6
DOIs
Publication statusPublished - 2002

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ASJC Scopus subject areas

  • Pharmacology (medical)

Cite this

Vessillier, S., Docquier, J. D., Rival, S., Frere, J. M., Galleni, M., Amicosante, G., Rossolini, G. M., & Franceschini, N. (2002). Overproduction and biochemical characterization of the Chryseobacterium meningosepticum B1aB metallo-β-lactamase. Antimicrobial Agents and Chemotherapy, 46(6), 1921-1927. https://doi.org/10.1128/AAC.46.6.1921-1927.2002