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             <jats:p>\n                      We aimed to characterize OXA-1054, a novel carbapenem-hydrolyzing class D \u03b2-lactamase (CHDL) detected in a multidrug-resistant\n                      <jats:italic toggle=\"yes\">Pseudomonas aeruginosa<\/jats:italic>\n                      clinical isolate. Antimicrobial susceptibility was determined by broth microdilution, and carbapenemase activity was confirmed by hydrolysis assays. Whole-genome sequencing via Illumina and PacBio platforms enabled comprehensive analysis of the resistome and plasmid architecture. The\n                      <jats:italic toggle=\"yes\">bla<\/jats:italic>\n                      <jats:sub>OXA-1054<\/jats:sub>\n                      gene was cloned in parallel with\n                      <jats:italic toggle=\"yes\">bla<\/jats:italic>\n                      <jats:sub>OXA-48<\/jats:sub>\n                      and\n                      <jats:italic toggle=\"yes\">bla<\/jats:italic>\n                      <jats:sub>OXA-198<\/jats:sub>\n                      into the pUCP24 plasmid. \u03b2-Lactamases were produced in\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      PAO1 for comparative evaluation of the phenotypic impact of each. OXA-48, OXA-198, and OXA-1054 \u03b2-lactamases were purified and further subjected to steady-state kinetic analysis, and 50% inhibitory activity of \u03b2-lactamase inhibitors was determined. The clinical\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      isolate ARGA00461 showed resistance to carbapenems and most \u03b2-lactam\/\u03b2-lactamase inhibitor combinations and tested positive in carbapenem hydrolysis assays. Genomic analysis revealed that the\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      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\n                      <jats:italic toggle=\"yes\">bla<\/jats:italic>\n                      <jats:sub>OXA-1054<\/jats:sub>\n                      gene was located in a small (\u22485 kbp) non-conjugative plasmid coexisting with a conjugative IncP plasmid, which likely facilitated its mobilization. Production of OXA-1054 in\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      PAO1 conferred a broader spectrum of \u03b2-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 \u03b2-lactam resistance in\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      , including carbapenems and \u03b2-lactam\/\u03b2-lactamase inhibitor combinations.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aac.01805-25","type":"journal-article","created":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T06:46:42Z","timestamp":1780037202000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Genetic and biochemical characterization of OXA-1054, a carbapenem-hydrolyzing class D \u03b2-lactamase conferring broad-spectrum \u03b2-lactam resistance in\n                    <i>Pseudomonas aeruginosa<\/i>"],"prefix":"10.1128","volume":"70","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8387-3784","authenticated-orcid":true,"given":"Luc\u00eda","family":"Gonz\u00e1lez-Pinto","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01qckj285","id-type":"ROR","asserted-by":"publisher"}],"name":"Servicio de Microbiolog\u00eda Cl\u00ednica & Grupo de Investigaci\u00f3n en Microbiolog\u00eda. Instituto de Investigaci\u00f3n Biom\u00e9dica de A Coru\u00f1a (INIBIC), Complexo Hospitalario Universitario de A Coru\u00f1a (CHUAC), SERGAS, Universidade da Coru\u00f1a (UDC)","place":["A Coru\u00f1a, Spain"]},{"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"publisher"}],"name":"CIBER de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III","place":["Madrid, Espa\u00f1a"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3476-9737","authenticated-orcid":true,"given":"Laura","family":"Monge-Olivares","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"publisher"}],"name":"CIBER de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III","place":["Madrid, Espa\u00f1a"]},{"id":[{"id":"https:\/\/ror.org\/03yxnpp24","id-type":"ROR","asserted-by":"publisher"}],"name":"Instituto de Biomedicina de Sevilla IBIS, Hospital Universitario Virgen Macarena\/CSIC\/Universidad de Sevilla","place":["Seville, Spain"]},{"id":[{"id":"https:\/\/ror.org\/03yxnpp24","id-type":"ROR","asserted-by":"publisher"}],"name":"Departamento de Microbiolog\u00eda, Facultad de Medicina, Universidad de Sevilla","place":["Seville, Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8039-6641","authenticated-orcid":true,"given":"Gloria","family":"P\u00e9rez-Rodr\u00edguez","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01qckj285","id-type":"ROR","asserted-by":"publisher"}],"name":"Servicio de Microbiolog\u00eda Cl\u00ednica & Grupo de Investigaci\u00f3n en Microbiolog\u00eda. Instituto de Investigaci\u00f3n Biom\u00e9dica de A Coru\u00f1a (INIBIC), Complexo Hospitalario Universitario de A Coru\u00f1a (CHUAC), SERGAS, Universidade da Coru\u00f1a (UDC)","place":["A Coru\u00f1a, Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6296-5949","authenticated-orcid":true,"given":"Pablo","family":"Aja-Macaya","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01qckj285","id-type":"ROR","asserted-by":"publisher"}],"name":"Servicio de Microbiolog\u00eda Cl\u00ednica & Grupo de Investigaci\u00f3n en Microbiolog\u00eda. 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