KPC-53 enzyme is a natural KPC variant which showed a duplication of L167E168 residues in the Omega-loop structure. The bla(K)(PC-53) gene was cloned both into pBC-SK and pET-24a vectors, and the recombinant plasmids were transferred by transformation in Escherichia coli competent cells to evaluate the antimicrobial susceptibility and to produce the enzyme. Compared to KPC-3, the KPC-53 was less stable and showed a dramatic reduction of k(cat) and K-cat/K-m versus several beta-lactams, in particular carbapenems. Indeed, a-2000-fold reduction was observed in the k(cat) values of KPC-53 for imipenem and meropenem. Concerning inhibitors, KPC-53 was susceptible to tazobactam and clavulanic acid but maintained resistance to avibactam. The molecular modeling indicates that the L167E168 duplication in KPC-53 modifies the interactions between residues involved in the catalytic pocket, changing the flexibility of the S2-loop, which is directly coupled with the catalytic properties of the KPC enzymes.

A Two Amino Acid Duplication, L167E168, in the Ω-Loop Drastically Decreases Carbapenemase Activity of KPC-53, a Natural Class A β-Lactamase / Piccirilli, Alessandra; Cherubini, Sabrina; Celenza, Giuseppe; Rossolini, Gian Maria; Brisdelli, Fabrizia; Segatore, Bernardetta; Principe, Luigi; Luzzaro, Francesco; Andriani, Lilia; Amicosante, Gianfranco; Perilli, Mariagrazia. - In: ANTIMICROBIAL AGENTS AND CHEMOTHERAPY. - ISSN 1098-6596. - ELETTRONICO. - 66:(2022), pp. 0-0. [10.1128/aac.02402-21]

A Two Amino Acid Duplication, L167E168, in the Ω-Loop Drastically Decreases Carbapenemase Activity of KPC-53, a Natural Class A β-Lactamase

Rossolini, Gian Maria;
2022

Abstract

KPC-53 enzyme is a natural KPC variant which showed a duplication of L167E168 residues in the Omega-loop structure. The bla(K)(PC-53) gene was cloned both into pBC-SK and pET-24a vectors, and the recombinant plasmids were transferred by transformation in Escherichia coli competent cells to evaluate the antimicrobial susceptibility and to produce the enzyme. Compared to KPC-3, the KPC-53 was less stable and showed a dramatic reduction of k(cat) and K-cat/K-m versus several beta-lactams, in particular carbapenems. Indeed, a-2000-fold reduction was observed in the k(cat) values of KPC-53 for imipenem and meropenem. Concerning inhibitors, KPC-53 was susceptible to tazobactam and clavulanic acid but maintained resistance to avibactam. The molecular modeling indicates that the L167E168 duplication in KPC-53 modifies the interactions between residues involved in the catalytic pocket, changing the flexibility of the S2-loop, which is directly coupled with the catalytic properties of the KPC enzymes.
66
0
0
Piccirilli, Alessandra; Cherubini, Sabrina; Celenza, Giuseppe; Rossolini, Gian Maria; Brisdelli, Fabrizia; Segatore, Bernardetta; Principe, Luigi; Luzzaro, Francesco; Andriani, Lilia; Amicosante, Gianfranco; Perilli, Mariagrazia
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in FLORE sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2158/1286780
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 0
  • ???jsp.display-item.citation.isi??? 0
social impact