A Kinetic Study of the Replacement by Site Saturation Mutagenesis of Residue 119 in NDM-1 Metallo-β-Lactamase.


Abstract

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 kcat/Km 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 kcat/Km values for ceftazidime about 25- and 89-fold higher, respectively, than that of NDM-1.

Submission Details

ID: CWvASvaZ3

Submitter: Shu-Ching Ou

Submission Date: March 20, 2019, 3:27 p.m.

Version: 1

Publication Details
Marcoccia F;Mercuri PS;Galleni M;Celenza G;Amicosante G;Perilli M,Antimicrob Agents Chemother (2018) A Kinetic Study of the Replacement by Site Saturation Mutagenesis of Residue 119 in NDM-1 Metallo-β-Lactamase. PMID:29784851
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
5ZJ1 2018-03-18T00:00:00+0000 0.96 Crystal structure of NDM-1 in complex with D-captopril derivative CYT-14
6C6I 2018-01-18T00:00:00+0000 1.65 Crystal structure of a chimeric NDM-1 metallo-beta-lactamase harboring the IMP-1 L3 loop
5ZJ2 2018-03-18T00:00:00+0000 1.1 Crystal structure of NDM-1 in complex with D-captopril
3PG4 2010-10-29T00:00:00+0000 2.0 The crystal structure of New Delhi Metallo-beta lactamase (NDM-1)
5ZJ7 2018-03-19T00:00:00+0000 0.96 Crystal structure of NDM-1 in complex with D-captopril derivative CY22
5ZJ8 2018-03-19T00:00:00+0000 0.96 Crystal structure of NDM-1 in complex with D-captopril derivative CY32
5ZGP 2018-03-10T00:00:00+0000 1.15 Crystal structure of NDM-1 at pH6.2 (Bis-Tris) in complex with hydrolyzed ampicillin
5ZH1 2018-03-10T00:00:00+0000 1.05 Crystal structure of NDM-1 at pH7.5 (Imidazole) with 2 molecules per asymmetric unit
6LJ7 2019-12-13T00:00:00+0000 1.16 Crystal structure of NDM-1 in complex with D-captopril derivative wss05010
3SBL 2011-06-05T00:00:00+0000 2.31 Crystal Structure of New Delhi Metal-beta-lactamase-1 from Klebsiella pneumoniae

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Metallo-beta-lactamase type 2 C7C422 BLAN1_KLEPN