Open AccessA Kinetic Study of the Replacement by Site Saturation Mutagenesis of Residue 119 in NDM-1 Metallo-β-Lactamase
Author(s) -
Francesca Marcoccia,
Paola Sandra Mercuri,
Moreno Galleni,
Giuseppe Celenza,
Gianfranco Amicosante,
Mariagrazia Perilli
Publication year - 2018
Publication title -
antimicrobial agents and chemotherapy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.07
H-Index - 259
eISSN - 1070-6283
pISSN - 0066-4804
DOI - 10.1128/aac.02541-17
Subject(s) - saturated mutagenesis , residue (chemistry) , chemistry , saturation (graph theory) , kinetic energy , mutagenesis , site directed mutagenesis , microbiology and biotechnology , environmental chemistry , biology , biochemistry , mutant , mathematics , physics , gene , combinatorics , quantum mechanics
New Delhi metallo-β-lactamase 1 (NDM-1) is a subclass B1 metallo-β-lactamase that exhibits a broad spectrum of activity against β-lactam antibiotics. Here we report the kinetic study of 6 Q119X variants obtained by site-directed mutagenesis of NDM-1. All Q119X variants were able to hydrolyze carbapenems, penicillins and first-, second-, third-, and fourth-generation cephalosporins very efficiently. In particular, Q119E, Q119Y, Q119V, and Q119K mutants showed improvements in k cat / K m values for penicillins, compared with NDM-1. The catalytic efficiencies of the Q119K variant for benzylpenicillin and carbenicillin were about 65- and 70-fold higher, respectively, than those of NDM-1. The Q119K and Q119Y enzymes had k cat / K m values for ceftazidime about 25- and 89-fold higher, respectively, than that of NDM-1.
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