title: Imipenem resistance in Klebsiella pneumoniae is associated with the combination of ACT-1, a plasmid-mediated AmpC beta-lactamase, and the foss of an outer membrane protein.
abstract: Six Escherichia coli and 12 Klebsiella pneumoniae isolates from a single hospital expressed a common beta-lactamase with a pI of approximately 9.0 and were resistant to cefoxitin and cefotetan (MIC ranges, 64 to > 128 and 16 to > 128 micrograms/ml, respectively). Seventeen of the 18 strains produced multiple beta-lactamases. Most significantly, three K. pneumoniae strains were also resistant to imipenem (MICs, 8 to 32 micrograms/ml). Spectrophotometric beta-lactamase assays with purified enzyme indicated hydrolysis of cephamycins, in addition to cephaloridine and benzylpenicillin. The 4ene encoding the pI 9.0 beta-lactamase (designated ACT-1 for AmpC type) was cloned and sequenced, which revealed an ampC-type beta-lactamase gene that originated from Enterobacter cloacae and that had 86% sequence homology to the P99 beta-lactamase and 94% homology to the partial sequence of MIR-1. Southern blotting revealed that the gene encoding ACT-1 was on a large plasmid in some of the K. pneumoniae strains as well as on the chromosomes of all of the strains, suggesting that the gene is located on an easily mobilized element. Outer membrane protein profiles of the K. pneumoniae strains revealed that the three imipenem-resistant strains were lacking a major outer membrane protein of approximately 42 kDa which was present in the imipenem-susceptible strains. ACT-1 is the first plasmid-mediated AmpC-type beta-lactamase derived from Enterobacter which has been completely sequenced. This work demonstrates that in addition to resistance to cephamycins, imipenem resistance can occur in K. pneumoniae when a high level of the ACT-1 beta-lactamase is produced in combination with the loss of a major outer membrane protein.
authors.affiliation: Wyeth-Ayerst Research, Pearl River, New York 10965, USA. bradfop@war.wyeth.com
authors.forename: P A
authors.initials: PA
authors.lastname: Bradford
authors.forename: C
authors.initials: C
authors.lastname: Urban
authors.forename: N
authors.initials: N
authors.lastname: Mariano
authors.forename: S J
authors.initials: SJ
authors.lastname: Projan
authors.forename: J J
authors.initials: JJ
authors.lastname: Rahal
authors.forename: K
authors.initials: K
authors.lastname: Bush
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000502:physiology; D015139:Blotting, Southern; D015780:Carbapenems / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000302:isolation & purification / Q000378:metabolism; D004196:Disease Outbreaks; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D004927:Escherichia coli Infections / Q000382:microbiology; D005260:Female; D006801:Humans; D015378:Imipenem / Q000494:pharmacology*; D007223:Infant; D007525:Isoelectric Focusing; D007710:Klebsiella Infections / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D018440:beta-Lactam Resistance / Q000235:genetics; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
chemical_list: D001425:Bacterial Outer Membrane Proteins; D015780:Carbapenems; D004269:DNA, Bacterial; D015378:Imipenem; D001618:beta-Lactamases