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 61 publications

(92 citation statements)

References 30 publications

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“…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

126

139

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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.

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Section: Resultssupporting

confidence: 90%

Class D β-Lactamases: Are They All Carbapenemases?

Antunes

Lamoureaux

Tóth

et al. 2014

Antimicrob Agents Chemother

126

139

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“…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

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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...

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“…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).…”

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confidence: 61%