Biochemical characterization of a cefotaximehydrolysing
β-lactamase encoded by a conjugative plasmid

Abstract

During the nosocomials infections occurring in the intensive care unit of the military hospital of Tunis in 2005, the Enterobacter cloacae BW 1150 strain was isolated from a stool culture. This strain was found to have a high level resistance to broad-spectrum -lactams. Resistance profile against the
various families of antibiotics was determined using the disc diffusion test. The minimal inhibitory concentrations values showed that this strain was resistant to the -lactams such as ampicillin and the extended spectrum cephalosporins (cefotaxime, ceftriaxon and cefpirome). Analysis of this strain by the disk diffusion test revealed synergies between amoxicillin-clavulanate (AMX-CA) and ceftriaxon, ceftazidime and cefotaxime. Cell sonicate of this isolate is very active against cefotaxime and showed a
specific activity (AS) of 7.54 U/mg for the same antibiotic. This activity was inhibited by the sulbactam and the clavulanic acid. Isoelectrofocusing methods revealed that the crude extract of the E. cloacae BW 1150 strain showed 1 - lactamase activity with an isoelectric piont (pI) of about 8. This activity was transferred by conjugation and was highly expressed in the transconjugant.