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 Franceschini, N.
Right arrow Articles by Rossolini, G. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Franceschini, N.
Right arrow Articles by Rossolini, G. M.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, December 2001, p. 3509-3516, Vol. 45, No. 12
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.12.3509-3516.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Characterization of OXA-29 from Legionella (Fluoribacter) gormanii: Molecular Class D beta -Lactamase with Unusual Properties

Nicola Franceschini,1 Letizia Boschi,2 Simona Pollini,2 Raphaël Herman,3 Mariagrazia Perilli,1 Moreno Galleni,3 Jean-Marie Frère,3 Gianfranco Amicosante,1 and Gian Maria Rossolini2,*

Dipartimento di Scienze e Tecnologie Biomediche, Università di L'Aquila, I-67100 L'Aquila,1 and Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, I-53100 Siena,2 Italy, and Laboratoire d'Enzymologie & Centre d'Ingénierie des Protéines, Institut de Chimie, Université de Liège, Sart Tilman, B-4000 Liège, Belgium3

Received 26 February 2001/Returned for modification 23 June 2001/Accepted 23 September 2001

A class D beta -lactamase determinant was isolated from the genome of Legionella (Fluoribacter) gormanii ATCC 33297T. The enzyme, named OXA-29, is quite divergent from other class D beta -lactamases, being more similar (33 to 43% amino acid identity) to those of groups III (OXA-1) and IV (OXA-9, OXA-12, OXA-18, and OXA-22) than to other class D enzymes (21 to 24% sequence identity). Phylogenetic analysis confirmed the closer ancestry of OXA-29 with members of the former groups. The OXA-29 enzyme was purified from an Escherichia coli strain overexpressing the gene via a T7-based expression system by a single ion-exchange chromatography step on S-Sepharose. The mature enzyme consists of a 28.5-kDa polypeptide and exhibits an isoelectric pH of >9. Analysis of the kinetic parameters of OXA-29 revealed efficient activity (kcat/Km ratios of >105 M-1 · s-1) for several penam compounds (oxacillin, methicillin, penicillin G, ampicillin, carbenicillin, and piperacillin) and also for cefazolin and nitrocefin. Oxyimino cephalosporins and aztreonam were also hydrolyzed, although less efficiently (kcat/Km ratios of around 103 M-1 · s-1). Carbapenems were neither hydrolyzed nor inhibitory. OXA-29 was inhibited by BRL 42715 (50% inhibitory concentration [IC50], 0.44 µM) and by tazobactam (IC50, 3.2 µM), but not by clavulanate. It was also unusually resistant to chloride ions (IC50, >100 mM). Unlike OXA-10, OXA-29 was apparently found as a dimer both in diluted solutions and in the presence of EDTA. Its activity was either unaffected or inhibited by divalent cations. OXA-29 is a new class D beta -lactamase that exhibits some unusual properties likely reflecting original structural and mechanistic features.

* Corresponding author. Mailing address: Dipartimento di Biologia Molecolare, Sez. di Microbiologia, Università di Siena, Policlinico "Le Scotte," 53100 Siena, Italy. Phone: 39 0577 233455. Fax: 39 0577 233325. E-mail: rossolini{at}unisi.it.

Antimicrobial Agents and Chemotherapy, December 2001, p. 3509-3516, Vol. 45, No. 12
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.12.3509-3516.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Jacoby, G. A. (2009). AmpC {beta}-Lactamases. Clin. Microbiol. Rev. 22: 161-182 [Abstract] [Full Text]  
  • Voha, C., Docquier, J.-D., Rossolini, G. M., Fosse, T. (2006). Genetic and Biochemical Characterization of FUS-1 (OXA-85), a Narrow-Spectrum Class D {beta}-Lactamase from Fusobacterium nucleatum subsp. polymorphum.. Antimicrob. Agents Chemother. 50: 2673-2679 [Abstract] [Full Text]  
  • Giuliani, F., Docquier, J.-D., Riccio, M. L., Pagani, L., Rossolini, G. M. (2005). OXA-46, a New Class D {beta}-Lactamase of Narrow Substrate Specificity Encoded by a blaVIM-1-Containing Integron from a Pseudomonas aeruginosa Clinical Isolate. Antimicrob. Agents Chemother. 49: 1973-1980 [Abstract] [Full Text]  
  • Poirel, L., Heritier, C., Nordmann, P. (2005). Genetic and biochemical characterization of the chromosome-encoded class B {beta}-lactamases from Shewanella livingstonensis (SLB-1) and Shewanella frigidimarina (SFB-1). J Antimicrob Chemother 55: 680-685 [Abstract] [Full Text]  
  • Girlich, D., Naas, T., Nordmann, P. (2004). Biochemical Characterization of the Naturally Occurring Oxacillinase OXA-50 of Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 48: 2043-2048 [Abstract] [Full Text]  
  • Toleman, M. A., Rolston, K., Jones, R. N., Walsh, T. R. (2003). Molecular and Biochemical Characterization of OXA-45, an Extended-Spectrum Class 2d' {beta}-Lactamase in Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 47: 2859-2863 [Abstract] [Full Text]  
  • Bellais, S., Girlich, D., Karim, A., Nordmann, P. (2002). EBR-1, a Novel Ambler Subclass B1 {beta}-Lactamase from Empedobacter brevis. Antimicrob. Agents Chemother. 46: 3223-3227 [Abstract] [Full Text]  
  • Avison, M. B., Simm, A. M. (2002). Sequence and genome context analysis of a new molecular class D {beta}-lactamase gene from Legionella pneumophila. J Antimicrob Chemother 50: 331-338 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»