Mutational effects on carbapenem hydrolysis of YEM-1, a new sub-class B2 metallo-β-lactamase from Yersinia mollaretii

The analysis of the genome sequence of Yersinia mollaretii (Y. mollaretii) ATCC 43969 indicates the presence of the blaYEM gene coding for YEM-1, a putative subclass B2 metallo-{beta}-lactamase. The objectives of our work were to produce, purify and complete the kinetic characterization of YEM-1. Compared to the known subclass B2 metallo-{beta}-lactamases, YEM-1 displayed a narrowest substrate profile since it is only able to hydrolyse imipenem with a high catalytic efficiency but not all the other carbapenems tested such as biapenem, meropenem, doripenem and ertapenem. A possible explanation of this peculiar activity profile is the presence of tyrosine 67 (loop L1), threonine 156 (loop L2) and serine 236 (loop L3) respectively. We showed that the substitution of Y67 broadened the activity profile of the enzyme for all carbapenems but still displayed a poor activity toward the other {beta}-lactam classes.