Role of a mutation at position 167 of CTX-M-19 in ceftazidime hydrolysis

Antimicrob Agents Chemother. 2004 May;48(5):1454-60. doi: 10.1128/AAC.48.5.1454-1460.2004.

Abstract

CTX-M-19 is a recently identified ceftazidime-hydrolyzing extended-spectrum beta-lactamase, which differs from the majority of CTX-M-type beta-lactamases that preferentially hydrolyze cefotaxime but not ceftazidime. To elucidate the mechanism of ceftazidime hydrolysis by CTX-M-19, the beta-lactam MICs of a CTX-M-19 producer, and the kinetic parameters of the enzyme were confirmed. We reconfirmed here that CTX-M-19 is also stable at a high enzyme concentration in the presence of bovine serum albumin (20 micro g/ml). Under this condition, we obtained more accurate kinetic parameters and determined that cefotaxime (k(cat)/K(m), 1.47 x 10(6) s(-1) M(-1)), cefoxitin (k(cat)/K(m), 62.2 s(-1) M(-1)), and aztreonam (k(cat)/K(m), 1.34 x 10(3) s(-1) M(-1)) are good substrates and that imipenem (k(+2)/K, 1.20 x 10(2) s(-1) M(-1)) is a poor substrate. However, CTX-M-18 and CTX-M-19 exhibited too high a K(m) value (2.7 to 5.6 mM) against ceftazidime to obtain their catalytic activity (k(cat)). Comparison of the MICs with the catalytic efficiency (k(cat)/K(m)) of these enzymes showed that some beta-lactams, including cefotaxime, ceftazidime, and aztreonam showed a similar correlation. Using the previously reported crystal structure of the Toho-1 beta-lactamase, which belongs to the CTX-M-type beta-lactamase group, we have suggested characteristic interactions between the enzymes and the beta-lactams ceftazidime, cefotaxime, and aztreonam by molecular modeling. Aminothiazole-bearing beta-lactams require a displacement of the aminothiazole moiety due to a severe steric interaction with the hydroxyl group of Ser167 in CTX-M-19, and the displacement affects the interaction between Ser130 and the acidic group such as carboxylate and sulfonate of beta-lactams.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aztreonam / pharmacology
  • Ceftazidime / metabolism*
  • Cephalosporins / metabolism*
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Hydrolysis
  • Kinetics
  • Microbial Sensitivity Tests
  • Models, Molecular
  • Monobactams / pharmacology
  • Mutagenesis, Site-Directed
  • Mutation / physiology*
  • beta-Lactamases / genetics*
  • beta-Lactamases / metabolism*

Substances

  • Cephalosporins
  • Monobactams
  • Ceftazidime
  • beta-Lactamases
  • beta-lactamase CTX-M-19
  • Aztreonam