title: TEM-80, a novel inhibitor-resistant beta-lactamase in a clinical isolate of Enterobacter cloacae.
abstract: Enterobacter cloacae Ecl261 was isolated with Escherichia coli Ec257 from the urine of a patient living in a nursing home. Both isolates were resistant to ticarcillin (MICs, 1,024 microg/ml), without significant potentiation of its activity by 2 microg of clavulanate per ml (MICs, 512 microg/ml), and susceptible to naturally active cephalosporins. This inhibitor-resistant phenotype was conferred in both strains by similar conjugative plasmids of 40 kb (Ecl261) and 30 kb (Ec257), which also conveyed resistance to sulfonamides and trimethoprim. Clinical and transconjugant strains produced a beta-lactamase with a pI of 5.2 which belonged to the TEM family, as indicated by specific PCR amplification. Compared with TEM-1, this enzyme exhibited lower catalytic efficiencies (14- and 120-fold less for amoxicillin and ticarcillin, respectively), and higher concentrations of beta-lactamase inhibitors were required to yield a 50% reduction in benzylpenicillin hydrolysis (750-, 82-, and 50-fold higher concentrations for clavulanate, sulbactam, and tazobactam, respectively). Gene sequencing revealed four nucleotide differences with the nucleotide sequence of bla(TEM-1A). The first replacement (T32C), located in the promoter region, was described as being responsible for the increase in the level of beta-lactamase production. The three other changes led to amino acid substitutions that define a new inhibitor-resistant TEM (IRT) beta-lactamase, TEM-80 (alternate name, IRT-24). Two of them, Met69Leu and Asn276Asp, have previously been related to inhibitor resistance. The additional mutation, Ile127Val, was demonstrated by site-directed mutagenesis to have a very weak effect, at least alone, on the IRT phenotype. This is the first description of an IRT beta-lactamase in E. cloacae. The horizontal transfer of bla(TEM-80) may have occurred either from Ec257 to Ecl261 or in the reverse order.
authors.affiliation: Laboratoire de Microbiologie, Université de Bordeaux 2, 33076 Bordeaux Cedex, France. corinne.arpin@bacterio.u-bordeaux2.fr
authors.forename: Corinne
authors.initials: C
authors.lastname: Arpin
authors.forename: Roger
authors.initials: R
authors.lastname: Labia
authors.forename: Véronique
authors.initials: V
authors.lastname: Dubois
authors.forename: Patrick
authors.initials: P
authors.lastname: Noury
authors.forename: Muriel
authors.initials: M
authors.lastname: Souquet
authors.forename: Claudine
authors.initials: C
authors.lastname: Quentin
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D004352:Drug Resistance, Microbial; D016972:Enterobacter cloacae / Q000187:drug effects / Q000201:enzymology*; D004756:Enterobacteriaceae Infections / Q000382:microbiology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism*; D047090:beta-Lactams / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D047090:beta-Lactams; D001618:beta-Lactamases; C461659:beta-lactamase TEM-80, Enterobacter cloacae