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Antimicrobial Agents and Chemotherapy, February 2001, p. 413-419, Vol. 45, No. 2
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.2.413-419.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Plasmid Location and Molecular Heterogeneity of the L1 and L2 beta -Lactamase Genes of Stenotrophomonas maltophilia

Matthew B. Avison,* Catherine S. Higgins, Charlotte J. von Heldreich, Peter M. Bennett, and Timothy R. Walsh

Bristol Centre for Antimicrobial Research and Evaluation (BCARE), Department of Pathology and Microbiology, School of Medical Sciences, University of Bristol, Bristol BS8 ITD, United Kingdom

Received 16 March 2000/Returned for modification 1 July 2000/Accepted 30 October 2000

An approximately 200-kb plasmid has been purified from clinical isolates of Stenotrophomonas maltophilia. This plasmid was found in all of the 10 isolates examined and contains both the L1 and the L2 beta -lactamase genes. The location of L1 and L2 on a plasmid makes it more likely that they could spread to other gram-negative bacteria, potentially causing clinical problems. Sequence analysis of the 10 L1 genes revealed three novel genes, L1c, L1d, and L1e, with 8, 12, and 20% divergence from the published strain IID 1275 L1 (L1a), respectively. The most unusual L1 enzyme (L1e) displayed markedly different kinetic properties, with respect to hydrolysis of nitrocefin and imipenem, compared to those of L1a (250- and 100-fold lower kcat/Km ratios respectively). L1c and L1d, in contrast, displayed levels of hydrolysis very similar to that of L1a. Several nonconservative amino acid differences with respect to L1a, L1b, L1c, and L1d were observed in the substrate binding-catalytic regions of L1e, and this could explain the kinetic differences. Three novel L2 genes (L2b, L2c, and L2d) were sequenced from the same isolates, and their sequences diverge from the published sequence of strain IID 1275 L2 (L2a) by 4, 9, and 25%, respectively. Differences in L1 and L2 gene sequences were not accompanied by similar divergences in 16S rRNA gene sequences, for which differences of <1% were found. It is therefore apparent that the L1 and L2 genes have evolved relatively quickly, perhaps because of their presence on a plasmid.

* Corresponding author. Mailing address: Department of Pathology and Microbiology, University of Bristol, School of Medical Sciences, University Walk, Bristol BS8 1TD, United Kingdom. Phone: (44) (117) 9287541. Fax: (44) (117) 9287896. E-mail: Matthewb.Avison{at}bris.ac.uk.

Antimicrobial Agents and Chemotherapy, February 2001, p. 413-419, Vol. 45, No. 2
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.2.413-419.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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