Interplay between OXA-10 β-Lactamase Production and Low Outer-Membrane Permeability in Carbapenem Resistance in Enterobacterales
Isaac Alonso-García,
Juan Carlos Vázquez-Ucha,
Marta Martínez-Guitián,
Cristina Lasarte-Monterrubio,
Salud Rodríguez-Pallares,
Pablo Camacho-Zamora,
Soraya Rumbo-Feal,
Pablo Aja-Macaya,
Lucía González-Pinto,
Michelle Outeda-García,
Romina Maceiras,
Paula Guijarro-Sánchez,
María José Muíño-Andrade,
Ana Fernández-González,
Marina Oviaño,
Concepción González-Bello,
Jorge Arca-Suárez,
Alejandro Beceiro,
Germán Bou
Affiliations
Isaac Alonso-García
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Juan Carlos Vázquez-Ucha
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Marta Martínez-Guitián
NANOBIOFAR, Centre for Research in Molecular Medicine and Chronic Diseases (CiMUS), Universidad de Santiago de Compostela, Campus Vida Avenida Barcelona s/n, 15782 Santiago de Compostela, Spain
Cristina Lasarte-Monterrubio
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Salud Rodríguez-Pallares
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Pablo Camacho-Zamora
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Soraya Rumbo-Feal
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Pablo Aja-Macaya
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Lucía González-Pinto
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Michelle Outeda-García
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Romina Maceiras
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Paula Guijarro-Sánchez
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
María José Muíño-Andrade
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Ana Fernández-González
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Marina Oviaño
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Concepción González-Bello
Centro Singular de Investigación en Química Biolóxica e Materiais Moleculares (CiQUS), Departamento de Química Orgánica, Universidade de Santiago de Compostela, Jenaro de la Fuente s/n, 15782 Santiago de Compostela, Spain
Jorge Arca-Suárez
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Alejandro Beceiro
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
Germán Bou
Servicio de Microbiología and Instituto de Investigación Biomédica A Coruña (INIBIC), Complexo Hospitalario Universitario A Coruña, As Xubias 84, 15006 A Coruña, Spain
The OXA-10 class D β-lactamase has been reported to contribute to carbapenem resistance in non-fermenting Gram-negative bacilli; however, its contribution to carbapenem resistance in Enterobacterales is unknown. In this work, minimum inhibitory concentrations (MICs), whole genome sequencing (WGS), cloning experiments, kinetic assays, molecular modelling studies, and biochemical assays for carbapenemase detection were performed to determine the impact of OXA-10 production on carbapenem resistance in two XDR clinical isolates of Escherichia coli with the carbapenem resistance phenotype (ertapenem resistance). WGS identified the two clinical isolates as belonging to ST57 in close genomic proximity to each other. Additionally, the presence of the blaOXA-10 gene was identified in both isolates, as well as relevant mutations in the genes coding for the OmpC and OmpF porins. Cloning of blaOXA-10 in an E. coli HB4 (OmpC and OmpF-deficient) demonstrated the important contribution of OXA-10 to increased carbapenem MICs when associated with porin deficiency. Kinetic analysis showed that OXA-10 has low carbapenem-hydrolysing activity, but molecular models revealed interactions of this β-lactamase with the carbapenems. OXA-10 was not detected with biochemical tests used in clinical laboratories. In conclusion, the β-lactamase OXA-10 limits the activity of carbapenems in Enterobacterales when combined with low permeability and should be monitored in the future.
