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In Vitro and In Silico Inhibition of Staphylococcus aureus Efflux Pump NorA by α-Pinene and Limonene

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Abstract

Since the discovery of the first antibiotics, bacteria have acquired a variety of resistance mechanisms, with efflux pump (EP) being the most prominent mechanism for intracellular targeting drugs. These proteins have become efficient mechanisms of resistance to antibiotics in species such as Staphylococcus aureus and, therefore, have been identified as promising therapeutic targets in antibacterial drug development. Accordingly, evidence suggests that monoterpenes can act as EP inhibitors and can be useful in circumventing bacterial resistance. This study aimed to evaluate the effectiveness of monoterpenes α-pinene and limonene as EP inhibitors against a strain of S. aureus expressing NorA protein. The minimum inhibitory concentration (MIC) against the 1199B strain of S. aureus, which carries genes encoding efflux proteins associated with antibiotic resistance to norfloxacin, was assessed through the broth microdilution method. The results obtained served as a subsidy for the analysis of the NorA pump inhibition with norfloxacin and ethidium bromide. Docking techniques, in silico, were used to evaluate the interaction of monoterpenes with NorA. Both monoterpenes showed no clinically effective antibacterial activity. Nevertheless, these compounds were found to decrease the MICs of ethidium bromide and norfloxacin indicating EP inhibition, which was confirmed by molecular docking analyses. In conclusion, α-pinene and limonene showed promising antibiotic-enhancing properties in S. aureus 1199B strain, indicating that monoterpenes can be used in targeted drug development to combat antibiotic resistance associated with EP expression.

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Funding

This research received no external funding.

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Authors and Affiliations

  1. Department of Biological Chemistry, Regional University of Cariri - URCA, Rua Cel. Antonio Luis 1161, Pimenta, Crato, CE, 63105-000, Brazil

    Ana Carolina Justino de Araújo, Priscilla Ramos Freitas, Cristina Rodrigues dos Santos Barbosa, Débora Feitosa Muniz, Ray Silva de Almeida, Irwin Rose Alencar de Menezes, Saulo Relison Tintino & Henrique Douglas Melo Coutinho

  2. Gonçalo Moniz Institute (IGM), Oswaldo Cruz Foundation (FIOCRUZ), Salvador, BA, Brazil

    Jaime Ribeiro-Filho

Authors
  1. Ana Carolina Justino de Araújo
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  2. Priscilla Ramos Freitas
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  3. Cristina Rodrigues dos Santos Barbosa
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  4. Débora Feitosa Muniz
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  5. Ray Silva de Almeida
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  6. Irwin Rose Alencar de Menezes
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  7. Jaime Ribeiro-Filho
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  8. Saulo Relison Tintino
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  9. Henrique Douglas Melo Coutinho
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Contributions

Conceptualization, A.C.J.A., J.R.-F., S.R.T., and H.D.M.C.; formal analysis, A.C.J.d.A., P.R.F., C.R.d.S.B., D.F.M., R.S.A., I.R.A.M., J.R.-F., and S.R.T.; funding acquisition, J.R.-F., S.R.T., and H.D.M.C.; investigation, A.C.J.d.A., P.R.F., C.R.d.S.B., D.F.M., R.S.A., I.R.A.M., J.R.-F., and S.R.T.; project administration, H.D.M.C.; resources, H.D.M.C.; supervision, ,J.R.-F., S.R.T., and H.D.M.C.; validation, A.C.J.A., J.R.-F. and S.R.T.; visualization, I.R.A.M., S.R.T., and H.D.M.C.; writing—original draft, A.C.J.d.A., P.R.F., C.R.d.S.B., R.S.A., and I.R.A.M.; writing—review & editing, S.R.T. and H.D.M.C. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Henrique Douglas Melo Coutinho.

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de Araújo, A.C.J., Freitas, P.R., dos Santos Barbosa, C.R. et al. In Vitro and In Silico Inhibition of Staphylococcus aureus Efflux Pump NorA by α-Pinene and Limonene. Curr Microbiol 78, 3388–3393 (2021). https://doi.org/10.1007/s00284-021-02611-9

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  • DOI: https://doi.org/10.1007/s00284-021-02611-9

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