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Engineering Biological Approaches for Detection of Toxic Compounds: A New Microbial Biosensor Based on the Pseudomonas putida TtgR Repressor

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Abstract

Environmental contamination by toxic organic compounds and antimicrobials is one of the causes for the recent surge of multidrug-resistant pathogenic bacteria. Monitoring contamination is therefore the first step in containment of antimicrobial resistance and requires the development of simple, sensitive, and quantitative tools that detect a broad spectrum of toxic compounds. In this study, we have engineered a new microbial biosensor based on the ttgR-regulated promoter that controls expression of the TtgABC extrusion efflux pump of Pseudomonas putida, coupled to a gfp reporter. The system was introduced in P. putida DOT-T1E, a strain characterized by its ability to survive in the presence of high concentrations of diverse toxic organic compounds. This whole-cell biosensor is capable to detect a wide range of structurally diverse antibiotics, as well as compounds such as toluene or flavonoids.

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Acknowledgments

We are grateful to M.D. Jesús Villar for the information provided on the antibiotics most commonly used in Hospitals against Pseudomonas. We thank Marisa Cañadas-Garre and Virgen de las Nieves Hospital for providing the antibiotics of clinical use. This work was supported by Spanish Ministry of Economy and Competitiveness, National Programme for Recruitment and Incorporation of Human Resources, Subprogramme: Ramon y Cajal RYC-2009-04570 and grant P11-CVI-7391 from Junta de Andalucía and EFDR.

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The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Department of Environmental Protection, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas (CSIC), C/Profesor Albareda, 1, 18008, Granada, Spain

    Manuel Espinosa-Urgel, Juan Luis Ramos & Ana María Fernández-Escamilla

  2. Systems Biology Research Unit, European Molecular Biology Laboratory (EMBL), Centre for Genomic Regulation, C/Dr. Aiguader, 88, 08003, Barcelona, Spain

    Luis Serrano

Authors
  1. Manuel Espinosa-Urgel
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  2. Luis Serrano
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  4. Ana María Fernández-Escamilla
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Corresponding author

Correspondence to Ana María Fernández-Escamilla.

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12033_2015_9849_MOESM1_ESM.tif

Supplementary material 1 (TIFF 5954 kb). Fig. S1. Toxic respond to toluene of the biosensor, via GFP expression, under UV-light illumination. A) In absence of toluene; B) In the presence of vapour phase of toluene

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Espinosa-Urgel, M., Serrano, L., Ramos, J.L. et al. Engineering Biological Approaches for Detection of Toxic Compounds: A New Microbial Biosensor Based on the Pseudomonas putida TtgR Repressor. Mol Biotechnol 57, 558–564 (2015). https://doi.org/10.1007/s12033-015-9849-2

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