Biochemical and molecular characterization of three new variants of AmpC beta-lactamases from Morganella morganii

Antimicrob Agents Chemother. 2006 Mar;50(3):962-7. doi: 10.1128/AAC.50.3.962-967.2006.

Abstract

Morganella morganii produces an inducible, chromosomally encoded AmpC beta-lactamase. We describe in this study three new variants of AmpC within this species with apparent pIs of 6.6 (M19 from M. morganii strain PP19), 7.4 (M29 from M. morganii strain PP29), and 7.8 (M37 from M. morganii strain PP37). After gene sequencing, deduced amino acid sequences displayed one to six substitutions when compared to the available Morganella AmpC sequences. An AmpR-encoding gene was also found upstream of ampC, including the LysR regulators' helix-turn-helix DNA-binding domain and the putative T-N11-A-protected region in the ampR-ampC intercistronic sequence. All three AmpC variants were purified from in vitro-generated derepressed mutants and showed overall similar kinetic parameters. None of the observed amino acid changes, occurring at the surface of the protein, appear to have a major influence in their catalytic properties. Morganella AmpCs exhibit the highest catalytic efficiencies (k(cat)/K(m)) on classical penicillins, cefoxitin, narrow-spectrum cephalosporins, and cefotaxime. Cefotaxime was more effectively hydrolyzed than other oxyimino-cephalosporins, whereas cefepime was 3 log-fold less efficiently hydrolyzed than other cephalosporins such as cephalothin. Several differences with other AmpC beta-lactamases were found. Ampicillin was more efficiently hydrolyzed than benzylpenicillin. High k(cat)/K(m) values were observed for oxacillin and piperacillin, which are usually poor substrates for AmpC. A fairly efficient hydrolysis of imipenem was detected as well. Aztreonam, carbenicillin, and tazobactam were effective transient inactivators of these variants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / isolation & purification
  • Base Sequence
  • Binding Sites
  • DNA / analysis
  • DNA / genetics
  • Genes, Bacterial
  • Genetic Variation*
  • Helix-Turn-Helix Motifs
  • Isoelectric Focusing
  • Isoelectric Point
  • Kinetics
  • Microbial Sensitivity Tests
  • Models, Molecular
  • Molecular Sequence Data
  • Morganella morganii / enzymology*
  • Morganella morganii / genetics
  • Morganella morganii / growth & development
  • Protein Conformation
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • beta-Lactamases / chemistry
  • beta-Lactamases / genetics*
  • beta-Lactamases / isolation & purification

Substances

  • Bacterial Proteins
  • DNA
  • AmpC beta-lactamases
  • beta-Lactamases