Evolution to carbapenem-hydrolyzing activity in noncarbapenemase class D β-lactamase OXA-10 by rational protein design

Luca, Filomena De; Benvenuti, Manuela; Carboni, Filippo; Pozzi, Cecilia; Rossolini, Gian María; Mangani, Stefano; Docquier, Jean Denis

doi:10.1073/pnas.1110530108

Cited by 64 publications

(92 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These kinetic parameters are similar to those reported previously for OXA-10, OXA-24/40, OXA-48, and OXA-58 (to our knowledge, kinetic parameters for OXA-2 and OXA-23 with oxacillin have not been published) (20,30,(32)(33)(34). The catalytic efficiency (k cat /K m ) of the enzymes with individual carbapenem substrates varied by as much as 3 orders of magnitude (ϳ10 3 to 10 6 M Ϫ1 s Ϫ1 ), with OXA-10 being the least efficient carbapenemase.…”

Section: Resultssupporting

confidence: 90%

Class D β-Lactamases: Are They All Carbapenemases?

Antunes

Lamoureaux

Tóth

et al. 2014

Antimicrob Agents Chemother

134

139

View full text Add to dashboard Cite

Carbapenem-hydrolyzing class D ␤-lactamases (CHDLs) are enzymes of the utmost clinical importance due to their ability to produce resistance to carbapenems, the antibiotics of last resort for the treatment of various life-threatening infections. The vast majority of these enzymes have been identified in Acinetobacter spp., notably in Acinetobacter baumannii. The OXA-2 and OXA-10 enzymes predominantly occur in Pseudomonas aeruginosa and are currently classified as narrow-spectrum class D ␤-lactamases. Here we demonstrate that when OXA-2 and OXA-10 are expressed in Escherichia coli strain JM83, they produce a narrow-spectrum antibiotic resistance pattern. When the enzymes are expressed in A. baumannii ATCC 17978, however, they behave as extended-spectrum ␤-lactamases and confer resistance to carbapenem antibiotics. Kinetic studies of OXA-2 and OXA-10 with four carbapenems have demonstrated that their catalytic efficiencies with these antibiotics are in the same range as those of some recognized class D carbapenemases. These results are in disagreement with the classification of the OXA-2 and OXA-10 enzymes as narrow-spectrum ␤-lactamases, and they suggest that other class D enzymes that are currently regarded as noncarbapenemases may in fact be CHDLs.

show abstract

Section: Resultssupporting

confidence: 90%

Class D β-Lactamases: Are They All Carbapenemases?

Antunes

Lamoureaux

Tóth

et al. 2014

Antimicrob Agents Chemother

134

139

View full text Add to dashboard Cite

show abstract

“…This result agrees with crystal structure analysis of OXA-48 showing that Arg 214 (which is part of a ␤5 strand) is critical for carbapenemase activity (21). In addition, recent studies point out the crucial role of this short loop connecting ␤5 and ␤6 strands in conferring carbapenemase activity of Ambler class D ␤-lactamases (22,23).…”

Section: Discussionsupporting

confidence: 80%

Genetic and Biochemical Characterization of OXA-405, an OXA-48-Type Extended-Spectrum β-Lactamase without Significant Carbapenemase Activity

Dortet

Oueslati

Jeannot

et al. 2015

Antimicrob Agents Chemother

View full text Add to dashboard Cite

The epidemiology of carbapenemases worldwide is showing that OXA-48 variants are becoming the predominant carbapenemase type in Enterobacteriaceae in many countries. However, not all OXA-48 variants possess significant activity toward carbapenems (e.g., OXA-163). Two Serratia marcescens isolates with resistance either to carbapenems or to extended-spectrum cephalosporins were successively recovered from the same patient. A genomic comparison using pulsed-field gel electrophoresis and automated Rep-PCR typing identified a 97.8% similarity between the two isolates. Both strains were resistant to penicillins and first-generation cephalosporins. The first isolate was susceptible to expanded-spectrum cephalosporins, was resistant to carbapenems, and had a significant carbapenemase activity (positive Carba NP test) related to the expression of OXA-48. The second isolate was resistant to expanded-spectrum cephalosporins, was susceptible to carbapenems, and did not express a significant imipenemase activity, (negative for the Carba NP test) despite possessing a bla OXA-48 -type gene. Sequencing identified a novel OXA-48-type ␤-lactamase, OXA-405, with a four-amino-acid deletion compared to OXA-48. The bla OXA-405 gene was located on a ca. 46-kb plasmid identical to the prototype IncL/M bla OXA-48 -carrying plasmid except for a ca. 16.4-kb deletion in the tra operon, leading to the suppression of self-conjugation properties. Biochemical analysis showed that OXA-405 has clavulanic acid-inhibited activity toward expanded-spectrum activity without significant imipenemase activity. This is the first identification of a successive switch of catalytic activity in OXA-48-like ␤-lactamases, suggesting their plasticity. Therefore, this report suggests that the first-line screening of carbapenemase producers in Enterobacteriaceae may be based on the biochemical detection of carbapenemase activity in clinical settings.A mbler class D ␤-lactamases (oxacillinases) are widely disseminated among clinically relevant Gram-negative bacteria (1). They exhibit a high degree of diversity of hydrolysis activity ranging from narrow to broad-spectrum hydrolysis activity toward ␤-lactams (1). Among the class D ␤-lactamases, several enzymes hydrolyze carbapenems. Most carbapenemhydrolyzing class D ␤-lactamases (CHDLs) are from Acinetobacter spp. (e.g., OXA-23, OXA-40, OXA-58, OXA-143) (2, 3), whereas OXA-48-type enzymes are identified in Enterobacteriaceae only (4). The OXA-48-derived CHDLs have initially been identified in Turkey (5), first in Klebsiella pneumoniae and then in other enterobacterial species (4). The known OXA-48 variants are currently as follows: (i) OXA-162, identified from K. pneumoniae isolates in Turkey (6); (ii) OXA-163, identified from K. pneumoniae and Enterobacter cloacae isolates in Argentina (7, 8); (iii) OXA-181, identified from a K. pneumoniae isolate in India (9); (iv) OXA-204, identified from K. pneumoniae isolates in patients having a link with North Africa (10); (v) OXA-232, identified in France from a K. pneumoni...

show abstract

“…A similar substrate expansion is observed in OXA-23 or OXA-24 clinical variants with a proline-to-serine substitution in the same loop (20). The sequence and length of the ␤5␤6 loop have been implicated in carbapenem targeting as well, as replacement of the ␤5␤6 loop from the narrow-spectrum OXA-10 with the loop from OXA-48 imparts carbapenemase activity to the former (34). Other investigations into the mechanism by which class D ␤-lactamases hydrolyze carbapenems have focused on a highly conserved leucine (L168 in OXA-24/40) and how its side chain affects the ability of a water molecule to enter the active site and adopt the correct orientation for efficient deacylation of the carbapenem acyl-intermediate (16,23).…”

mentioning

confidence: 61%