title: Transferable production of PER-1 beta-lactamase in Pseudomonas aeruginosa.
abstract: PER-1, an extended-spectrum class A beta-lactamase, has been described only from Pseudomonas aeruginosa RNL-1, which was obtained in France in 1991. During studies on ceftazidime-resistant P. aeruginosa collected from December 1991 to July 1993 in Ankara, Turkey, we found 14 further isolates with PER-1 enzyme, as recognised by isoelectric point (pI 5.4), hydrolytic activity, gene hybridization and DNA sequence. Five of these isolates also had OXA-10 (PSE-2)-related enzymes and one hyper-produced ampC beta-lactamase, whereas eight had PER-1 enzyme alone. The last group, from four wards, appeared to be identical, giving the same DNA restriction patterns and carrying the PER-1 gene on an 8.5 kb HincII fragment. Two more producers were related but the other four were unique. In several representatives of the group of eight replicates, the PER-1 gene was shown to be encoded on a plasmid, larger than 154 kb in size, which transferred to P. aeruginosa PU21. A further isolate had the gene on an even larger conjugative plasmid. By contrast, the PER-1 gene reportedly was chromosomally-inserted in strain RNL-1. The PER-1 producers and their transconjugants were highly resistant to ceftazidime and aztreonam (MIC > or = 128 mg/L) but not to carbapenems or latamoxet. Piperacillin insusceptibility was marginal (MIC 8 mg/L). Clavulanate 4 mg/L, but not tazobactam 4 mg/L, reversed resistance to ceftazidime and carbenicillin. Purification of the enzyme to homogeneity was achieved by three ion exchanges and one gel filtration. We found much lower Vmax rates for aminothiazolyl cephalosporins than reported previously for PER-1 enzyme. This reflected the present assays being in 0.1 M phosphate buffer pH 7.0, whereas the previous were pH-stat-regulated; concentrated phosphate reduced enzyme activity against ceftazidime, but not against cephaloridine.
authors.affiliation: Department of Medical Microbiology, London Hospital Medical College, UK.
authors.forename: F
authors.initials: F
authors.lastname: Danel
authors.forename: L M
authors.initials: LM
authors.lastname: Hall
authors.forename: D
authors.initials: D
authors.lastname: Gur
authors.forename: H E
authors.initials: HE
authors.lastname: Akalin
authors.forename: D M
authors.initials: DM
authors.lastname: Livermore
mesh_terms: D001483:Base Sequence; D002442:Ceftazidime / Q000494:pharmacology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis; D004352:Drug Resistance, Microbial; D006801:Humans; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D002442:Ceftazidime; D001618:beta-Lactamases