Emergence of Oxacillinase-Mediated Resistance to Imipenem in Klebsiella pneumoniae

Klebsiella pneumoniae strain 11978 was isolated in Turkey in 2001 and was found to be resistant to all β-lactams, including carbapenems. Cloning and expression in Escherichia coli identified five β-lactamases, including two novel oxacillinases. The β-lactamase OXA-48 hydrolyzed imipenem at a high level and was remotely related (less than 46% amino acid identity) to the other oxacillinases. It hydrolyzed penicillins and imipenem but not expanded-spectrum cephalosporins. The blaOXA-48 gene was plasmid encoded and not associated with an integron, in contrast to most of the oxacillinase genes. An insertion sequence, IS1999, was found immediately upstream of blaOXA-48. Another plasmid that encoded a second oxacillinase gene, blaOXA-47, located inside a class 1 integron was identified in K. pneumoniae 11978. OXA-47 had a narrow spectrum of hydrolysis activity and did not hydrolyze ceftazidime or imipenem, as is found for the β-lactamase (OXA-1) to which it is related. In addition, β-lactamases TEM-1 and SHV-2a were expressed from the same K. pneumoniae isolate. Analysis of the outer membrane proteins of this isolate revealed that it lacked a porin of ca. 36 kDa. Thus, the high-level resistance to β-lactams of this clinical isolate resulted from peculiar β-lactamases and modification of outer membrane proteins.