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Antimicrobial Agents and Chemotherapy, July 2007, p. 2359-2365, Vol. 51, No. 7
0066-4804/07/$08.00+0     doi:10.1128/AAC.01395-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Biochemical Characterization of PER-2 and Genetic Environment of blaPER-2{triangledown}

Pablo Power,1 José Di Conza,2 María Margarita Rodríguez,1 Bárbara Ghiglione,1 Juan A. Ayala,3 José María Casellas,4 Marcela Radice,1 and Gabriel Gutkind1*

Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina,1 Cátedra de Microbiología General, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina,2 Centro de Biología Molecular "Severo Ochoa," Madrid, Spain,3 Centro de Estudios en Antimicrobianos, Martínez, Argentina4

Received 7 November 2006/ Returned for modification 20 December 2006/ Accepted 6 April 2007

PER-2 was the first detected and the second most prevalent extended-spectrum β-lactamase in clinical pathogens isolated in Argentina and was also reported only in other South American countries. Citrobacter freundii 33587 was isolated in 1999 in Buenos Aires and was resistant to all tested β-lactams except cephamycins and carbapenems. The strain produced both plasmid-borne TEM-1 and PER-2 (pI 5.4), which could be transferred by conjugation. By PCR screening, thermal asymmetric interlaced PCR, and DNA sequencing, we detected an ISPa12/IS1387a insertion sequence upstream of blaPER-2, previously reported as also being associated with blaPER-1. The presence of similar structures upstream of blaPER-1 and blaPER-2 suggests a common origin and mobilization. Compared to blaPER-1 genes, an additional putative promoter for blaPER-2 was found. PER-2 kinetic analysis showed its high hydrolysis efficiencies toward both CTX and CAZ (kcat/Km, 0.76 and 0.43 µM–1·s–1, respectively).

* Corresponding author. Mailing address: Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina. Phone: 54 11 4964 8285. Fax: 54 11 4508 3645. E-mail: ggutkind{at}ffyb.uba.ar

{triangledown} Published ahead of print on 16 April 2007.

Antimicrobial Agents and Chemotherapy, July 2007, p. 2359-2365, Vol. 51, No. 7
0066-4804/07/$08.00+0     doi:10.1128/AAC.01395-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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