Biochemical Characterization of PER-2 and Genetic Environment of blaPER-2
Abstract
Biochemical Characterization of PER-2 and Genetic Environment of bla PER-2 ▿ 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 Gutkind 1 , * 1 Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina 2 Cátedra de Microbiología General, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina 3 Centro de Biología Molecular “Severo Ochoa,” Madrid, Spain 4 Centro de Estudios en Antimicrobianos, Martínez, Argentina ABSTRACT 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 IS Pa12 /IS 1387a insertion sequence upstream of bla PER-2 , previously reported as also being associated with bla PER-1 . The presence of similar structures upstream of bla PER-1 and bla PER-2 suggests a common origin and mobilization. Compared to bla PER-1 genes, an additional putative promoter for bla PER-2 was found. PER-2 kinetic analysis showed its high hydrolysis efficiencies toward both CTX and CAZ ( k cat / K m , 0.76 and 0.43 μM −1 ·s −1 , respectively).