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Antimicrobial Agents and Chemotherapy, August 2002, p. 2427-2434, Vol. 46, No. 8
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.8.2427-2434.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Characterization of a Novel Plasmid-Mediated Cephalosporinase (CMY-9) and Its Genetic Environment in an Escherichia coli Clinical Isolate

Yohei Doi,* Naohiro Shibata, Keigo Shibayama, Kazunari Kamachi, Hiroshi Kurokawa, Keiko Yokoyama, Tetsuya Yagi, and Yoshichika Arakawa

Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious Diseases, Tokyo, Japan

Received 19 November 2001/ Returned for modification 9 February 2002/ Accepted 3 May 2002

An Escherichia coli strain, HKYM68, which showed resistance to broad-spectrum cephalosporins was isolated from a sputum specimen in Japan. The high-level resistance of the strain to ceftazidime, cefpirome, and moxalactam was carried by a self-transferable plasmid. The ß-lactamase gene responsible for the resistance was cloned and sequenced. The deduced amino acid sequence of this gene product, CMY-9, had a single amino acid substitution (E85D), the residue reported to be part of the recognition site for the R1 side chain of ß-lactams, compared with the amino acid sequence of CMY-8 and also had 78% identity with the amino acid sequence of CepH, a chromosomal cephalosporinase of Aeromonas hydrophila. A sul1-type class 1 integron containing an aacA1-orfG gene cassette was identified upstream of blaCMY-9 and ended with a truncated 3' conserved segment. The following 2.1 kb was almost identical to the common region of integrons In6 and In7 and the integron of pSAL-1, except that orf513 encoding a putative transposase was identified instead of orf341 due to addition of a single nucleotide. blaCMY-9 was closely located downstream of the end of the common region. These observations are indicative of the exogenous derivation of blaCMY-9 from some environmental microorganisms such as aeromonads.

* Corresponding author. Mailing address: Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan. Phone: 81-42-561-0771, ext. 538. Fax: 81-42-561-7173. E-mail: yohei{at}nih.go.jp.

Antimicrobial Agents and Chemotherapy, August 2002, p. 2427-2434, Vol. 46, No. 8
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.8.2427-2434.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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