Genetic and biochemical characterization of OXA-1054, a carbapenem-hydrolyzing class D β-lactamase conferring broad-spectrum β-lactam resistance in <i>Pseudomonas aeruginosa</i>

González-Pinto, Lucía;Monge-Olivares, Laura;Pérez-Rodríguez, Gloria;Aja-Macaya, Pablo;Arca-Suárez, Jorge;et.al.
(2026) Antimicrobial Agents and Chemotherapy — Vol. 70, n° 7, p. e0180525 (2026)

Files

González-Pinto L, Antimicrob Agents Chemother, 2026.pdf
  • Open Access
  • Adobe PDF
  • 685.95 KB
Download

Details

Authors
  • González-Pinto, Lucíaorcid-logoServicio de Microbiología Clínica & Grupo de Investigación en Microbiología. Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC)
    Author
  • Monge-Olivares, Lauraorcid-logoCIBER de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III
    Author
  • Pérez-Rodríguez, Gloriaorcid-logoServicio de Microbiología Clínica & Grupo de Investigación en Microbiología. Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC)
    Author
  • Aja-Macaya, Pabloorcid-logoServicio de Microbiología Clínica & Grupo de Investigación en Microbiología. Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC)
    Author
  • Bogaerts, PierreUCLouvain
    Author
  • Arca-Suárez, Jorgeorcid-logoServicio de Microbiología Clínica & Grupo de Investigación en Microbiología. Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC)
    Author
Show more
Abstract
We aimed to characterize OXA-1054, a novel carbapenem-hydrolyzing class D β-lactamase (CHDL) detected in a multidrug-resistant Pseudomonas aeruginosa clinical isolate. Antimicrobial susceptibility was determined by broth microdilution, and carbapenemase activity was confirmed by hydrolysis assays. Whole-genome sequenc ing via Illumina and PacBio platforms enabled comprehensive analysis of the resis tome and plasmid architecture. The bla OXA-1054 gene was cloned in parallel with bla OXA-48 and bla OXA-198 into the pUCP24 plasmid. β-Lactamases were produced in P. aeruginosa PAO1 for comparative evaluation of the phenotypic impact of each. OXA-48, OXA-198, and OXA-1054 β-lactamases were purified and further subjected to steady-state kinetic analysis, and 50% inhibitory activity of β-lactamase inhibitors was determined. The clinical P. aeruginosa isolate ARGA00461 showed resistance to carbapenems and most β-lactam/β-lactamase inhibitor combinations and tested positive in carbapenem hydrolysis assays. Genomic analysis revealed that the P. aeruginosa isolate ARGA00461 carried a gene coding for a previously uncharacterized CHDL, designated OXA-1054, which is closely related to the OXA-372 enzyme of environmental origin. The bla OXA-1054 gene was located in a small (≈5 kbp) non-conjugative plasmid coexisting with a conjugative IncP plasmid, which likely facilitated its mobilization. Production of OXA-1054 in P. aeruginosa PAO1 conferred a broader spectrum of β-lactam resistance than other CHDLs. Enzyme kinetics confirmed carbapenem hydrolysis with catalytic efficiency comparable to OXA-48 and revealed higher affinity for cefepime compared to OXA-48 and OXA-198. Among the inhibitors tested, only avibactam demonstrated relevant inhibitory activity. OXA-1054 mediates broad-spectrum β-lactam resistance in P. aeruginosa, including carbapenems and β-lactam/β-lactamase inhibitor combinations.
Affiliations

Citations

  • APA
  • Chicago
  • FWB

González-Pinto, L., Monge-Olivares, L., Pérez-Rodríguez, G., Aja-Macaya, P., Sánchez-Peña, L., Gomis-Font, M., Rodríguez-Pallares, S., Blanco-Martín, T., López-Cerero, L., Beceiro, A., Bogaerts, P., Oliver, A., Bou, G., & Arca-Suárez, J. (2026). Genetic and biochemical characterization of OXA-1054, a carbapenem-hydrolyzing class D β-lactamase conferring broad-spectrum β-lactam resistance in <i>Pseudomonas aeruginosa</i>. Antimicrobial Agents and Chemotherapy, 70(7), e0180525. https://doi.org/10.1128/aac.01805-25 (Original work published 2026)