AAC
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Vessillier, S.
Right arrow Articles by Franceschini, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Vessillier, S.
Right arrow Articles by Franceschini, N.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, June 2002, p. 1921-1927, Vol. 46, No. 6
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.6.1921-1927.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Overproduction and Biochemical Characterization of the Chryseobacterium meningosepticum BlaB Metallo-ß-Lactamase

Sandrine Vessillier,1 Jean-Denis Docquier,2 Sandrine Rival,3 Jean-Marie Frere,4 Moreno Galleni,4 Gianfranco Amicosante,1 Gian Maria Rossolini,2 and Nicola Franceschini1*

Dipartimento di Scienze e Tecnologie Biomediche, Università di L'Aquila, I-67100 L'Aquila,1 Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, I-53100 Siena, Italy,2 Institut für Biochemie, Gebande 6, Universität des Saarlandes, 66123 Saarbrücken, Germany,3 Laboratoire d'Enzymologie and Centre d'Ingénierie des Protéines, Institut de Chimie, Université de Liège, Sart Tilman, B-4000 Liège, Belgium4

Received 31 July 2001/ Returned for modification 23 November 2001/ Accepted 16 March 2002

The BlaB metallo-ß-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-{alpha}-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-ß-lactamase inhibitors, including ß-iodopenicillanate, sulbactam and, although less efficiently, tazobactam.

* Corresponding author. Mailing address: Dipartimento di Scienze e Tecnologie Biomediche, Università dell'Aquila, Loc. Coppito, 67100-L'Aquila, Italy. Phone: 39 0862 433456. Fax: 39 0862 433433. E-mail: nicola.franceschini{at}cc.univaq.it.

Antimicrobial Agents and Chemotherapy, June 2002, p. 1921-1927, Vol. 46, No. 6
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.6.1921-1927.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Clin. Vaccine Immunol. Clin. Microbiol. Rev.
J. Clin. Microbiol. ALL ASM JOURNALS

Copyright © 2002 by the American Society for Microbiology. All rights reserved.