Biochemical Characterization of Sfh-I, a Subclass B2 Metallo-β-Lactamase from Serratia fonticola UTAD54
AbstractBiochemical Characterization of Sfh-I, a Subclass B2 Metallo-β-Lactamase from Serratia fonticola UTAD54 ▿ Fátima Fonseca 1 , 2 , * , Christopher J. Arthur 3 , Elizabeth H. C. Bromley 3 , † , Bart Samyn 4 , Pablo Moerman 4 , Maria José Saavedra 5 , António Correia 1 and James Spencer 2 , * 1 CESAM & Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal 2 School of Cellular and Molecular Medicine, University of Bristol Medical Sciences Building, University Walk, Bristol BS8 1TD, United Kingdom 3 School of Chemistry, University of Bristol, Cantock's Close, Bristol BS8 1TS, United Kingdom 4 Laboratory of Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry, Physiology and Microbiology, Ghent University, Gent, Belgium 5 CECAV, Department of Veterinary Science, University of Trás-os-Montes e Alto Douro, 5001-801 Vila Real, Portugal ABSTRACT The subclass B2 metallo-β-lactamase (MBL) Sfh-I from Serratia fonticola UTAD54 was cloned and overexpressed in Escherichia coli . The recombinant protein binds one equivalent of zinc, as shown by mass spectrometry, and preferentially hydrolyzes carbapenem substrates. However, compared to other B2 MBLs, Sfh-I also shows limited hydrolytic activity against some additional substrates and is not inhibited by a second equivalent of zinc. These data confirm Sfh-I to be a subclass B2 metallo-β-lactamase with some distinctive properties.