title: Plasmid location and molecular heterogeneity of the L1 and L2 beta-lactamase genes of Stenotrophomonas maltophilia.
abstract: 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 k(cat)/K(m) 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.
authors.affiliation: 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. Mathewb.Avison@bris.ac.uk
authors.forename: M B
authors.initials: MB
authors.lastname: Avison
authors.forename: C S
authors.initials: CS
authors.lastname: Higgins
authors.forename: C J
authors.initials: CJ
authors.lastname: von Heldreich
authors.forename: P M
authors.initials: PM
authors.lastname: Bennett
authors.forename: T R
authors.initials: TR
authors.lastname: Walsh
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D004269:DNA, Bacterial / Q000235:genetics; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020588:Stenotrophomonas / Q000737:chemistry / Q000235:genetics*; D001618:beta-Lactamases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D007527:Isoenzymes; C088032:beta-lactamase L1; C425104:beta-lactamase L2; D001618:beta-Lactamases