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Synthesis and electrochemical detection of a thiazolyl-indole natural product isolated from the nosocomial pathogen Pseudomonas aeruginosa

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Abstract

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen, capable of surviving in a broad range of natural environments and quickly acquiring resistance. It is associated with hospital-acquired infections, particularly in patients with compromised immunity, and is the primary cause of morbidity and mortality in cystic fibrosis (CF) patients. P. aeruginosa is also of nosocomial importance on dairy farms and veterinary hospitals, where it is a key morbidity factor in bovine mastitis. P. aeruginosa uses a cell-cell communication system consisting of signalling molecules to coordinate bacterial secondary metabolites, biofilm formation, and virulence. Simple and sensitive methods for the detection of biomolecules as indicators of P. aeruginosa infection would be of great clinical importance. Here, we report the synthesis of the P. aeruginosa natural product, barakacin, which was recently isolated from the bovine ruminal strain ZIO. A simple and sensitive electrochemical method was used for barakacin detection using a boron-doped diamond (BDD) and glassy carbon (GC) electrodes, based on cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The influence of electrolyte pH on the peak potential and peak currents was also investigated. At pH 2.0, the peak current was linearly dependent on barakacin concentration (in the range used, 1–10 μM), with correlation coefficients greater than 0.98 on both electrodes. The detection limit (S/N = 3) on the BDD electrode was 100-fold lower than that obtained on the GC electrode. The optimized method using the BDD electrode was extended to bovine (cow feces) and human (sputum of a CF patient) samples. Spiked barakacin was easily detected in these matrices at a limit of 0.5 and 0.05 μM, respectively.

Electrochemical detection of barakacin

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Acknowledgments

This research was financially supported by SFI/EI Technology Innovation Development Award (TIDA) (SFI/12/TIDA/B2405). FOG acknowledges the grants awarded by the European Commission (FP7-PEOPLE-2013-ITN, 607786; FP7-KBBE-2012-6, CP-TP-312184; FP7-KBBE-2012-6, 311975; OCEAN 2011–2, 287589; Marie Curie 256596; EU-634486); Science Foundation Ireland (SSPC-2, 12/RC/2275; 13/TIDA/B2625; 12/TIDA/B2411; 12/TIDA/B2405; 14/TIDA/2438); the Department of Agriculture and Food (FIRM/RSF/CoFoRD; FIRM 08/RDC/629; FIRM 1/F009/MabS; FIRM 13/F/516); the Irish Research Council for Science, Engineering and Technology (PD/2011/2414; GOIPG/2014/647); the Health Research Board/Irish Thoracic Society (MRCG-2014-6); the Marine Institute (Beaufort award C2CRA 2007/082); and Teagasc (Walsh Fellowship 2013). JDG thanks Science Foundation Ireland (08/SRC/B1412) for research funding of the Irish Separation Science Cluster (ISSC) under the Strategic Research Cluster Programme. GMG acknowledges supports by Science Foundation Ireland (SFI/12/IP/1315, SFI/09/RFP/CHS2353, and SSPC2 12/RC/2275), the Irish Research Council (GOIPG/2013/336), and the UCC for a Strategic Research Fund Ph.D. Studentship.

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Authors and Affiliations

  1. Innovative Chromatography Group, Irish Separation Science Cluster (ISSC), Cork, Ireland

    Alyah Buzid, Phyllis E. Hayes, Fengjun Shang, John H. T. Luong & Jeremy D. Glennon

  2. Department of Chemistry and Analytical & Biological Chemistry Research Facility (ABCRF), University College Cork, Cork, Ireland

    Alyah Buzid, Eoin Ó Muimhneacháin, Phyllis E. Hayes, Leticia M. Pardo, Fengjun Shang, John H. T. Luong, Jeremy D. Glennon & Gerard P. McGlacken

  3. BIOMERIT Research Centre, Department of Microbiology, University College Cork, Cork, Ireland

    F. Jerry Reen & Fergal O’Gara

  4. School of Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, Perth, WA, 6845, Australia

    Fergal O’Gara

  5. School of Chemical Sciences, University of Auckland, 23 Symonds Street, Auckland, New Zealand

    Jonathan Sperry

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  1. Alyah Buzid
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  2. Eoin Ó Muimhneacháin
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  9. John H. T. Luong
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Corresponding authors

Correspondence to Jeremy D. Glennon or Gerard P. McGlacken.

Ethics declarations

Sputum samples were collected from paediatric patients attending the CF clinic at Cork University Hospital, Ireland. Ethical approval was granted by the Clinical Research Ethics Committee (CREC) for sputum collection and samples were handled according to the approved guidelines. Written informed consent from all patients/guardians was obtained for acquisition and analysis outlined in this study.

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The authors declare that they have no competing interests.

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Buzid, A., Muimhneacháin, E.Ó., Reen, F.J. et al. Synthesis and electrochemical detection of a thiazolyl-indole natural product isolated from the nosocomial pathogen Pseudomonas aeruginosa . Anal Bioanal Chem 408, 6361–6367 (2016). https://doi.org/10.1007/s00216-016-9749-8

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