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Antimicrobial Agents and Chemotherapy, August 2001, p. 2269-2275, Vol. 45, No. 8
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.8.2269-2275.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Novel Cefotaximase (CTX-M-16) with Increased Catalytic Efficiency Due to Substitution Asp-240right-arrow Gly

R. Bonnet,1,* C. Dutour,1 J. L. M. Sampaio,2 C. Chanal,1 D. Sirot,1 R. Labia,3 C. De Champs,1 and J. Sirot1

Laboratoire de Bactériologie, Faculté de Médecine, 63001 Clermont-Ferrand Cedex,1 and UMR 175, CNRS-MNHN, 29000 Quimper,3 France, and Setor de Bacteriologia, Laboratório Lâmina LTDA, 71 - Botafogo, Rio de Janeiro, Brazil 22280-0302

Received 28 November 2000/Returned for modification 19 March 2001/Accepted 11 May 2001

Three clinical strains (Escherichia coli Rio-6, E. coli Rio-7, and Enterobacter cloacae Rio-9) collected in 1996 and 1999 from hospitals in Rio de Janeiro (Brazil) were resistant to broad-spectrum cephalosporins and gave a positive double-disk synergy test. Two blaCTX-M genes encoding beta -lactamases of pl 7.9 and 8.2 were implicated in this resistance: the blaCTX-M-9 gene observed in E. coli Rio-7 and E. cloacae Rio-9 and a novel CTX-M-encoding gene, designated blaCTX-M-16, observed in E. coli strain Rio-6. The deduced amino acid sequence of CTX-M-16 differed from CTX-M-9 only by the substitution Asp-240right-arrowGly. The CTX-M-16-producing E. coli transformant exhibited the same level of resistance to cefotaxime (MIC, 16 µg/ml) but had a higher MIC of ceftazidime (MIC, 8 versus 1 µg/ml) than the CTX-M-9-producing transformant. Enzymatic studies revealed that CTX-M-16 had a 13-fold higher affinity for aztreonam and a 7.5-fold higher kcat for ceftazidime than CTX-M-9, thereby showing that the residue in position 240 can modulate the enzymatic properties of CTX-M enzymes. The two blaCTX-M-9 genes and the blaCTX-M-16 gene were located on different plasmids, suggesting the presence of mobile elements associated with CTX-M-encoding genes. CTX-M-2 and CTX-M-8 enzymes were found in Brazil in 1996, and two other CTX-M beta -lactamases, CTX-M-9 and CTX-M-16, were subsequently observed. These reports are evidence of the diversity of CTX-M-type extended-spectrum beta -lactamases in Brazil.

* Corresponding author. Mailing address: Faculté de Médecine, Service de Bactériologie-Virologie, 28, Place Henri Dunant, 63 001 Clermont-Ferrand Cedex, France. Phone: 33 4 73 60 80 18. Fax: 33 4 73 27 74 94. E-mail: Richard.Bonnet{at}u-clermont1.fr.

Antimicrobial Agents and Chemotherapy, August 2001, p. 2269-2275, Vol. 45, No. 8
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.8.2269-2275.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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