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A rapid and reagent-free bioassay for the detection of dioxin-like compounds and other aryl hydrocarbon receptor (AhR) agonists using autobioluminescent yeast

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Abstract

An autonomously bioluminescent Saccharomyces cerevisiae BLYAhS bioreporter was developed in this study for the simple and rapid detection of dioxin-like compounds (DLCs) and aryl hydrocarbon receptor (AhR) agonists. This recombinant yeast reporter was based on a synthetic bacterial luciferase reporter gene cassette (lux) that can produce the luciferase as well as the enzymes capable of self-synthesizing the requisite substrates for bioluminescent production from endogenous cellular metabolites. As a result, bioluminescent signal production is generated continuously and autonomously without cell lysis or exogenous reagent addition. By linking the expression of the autobioluminescent lux reporter cassette to AhR activation via the use of a dioxin-responsive promoter, the S. cerevisiae BLYAhS bioreporter emitted a bioluminescent signal in response to DLC exposure in a dose-responsive manner. The model dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), could be detected within 4 h with a half maximal effective concentration (EC50) of ~ 8.1 nM and a lower detection limit of 500 pM. The autobioluminescent response of BLYAhS to other AhR agonists, including 2,3,7,8-tetrachlorodibenzofuran (TCDF), polychlorinated bisphenyl congener 126 (PCB-126) and 169 (PCB-169), 1,2,3,6,7,8-hexachlorodibenzo-p-dioxin (HxCDD), 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD), benzo[a]pyrene (BaP), and β-naphthoflavone (bNF), were also characterized in this study. The non-destructive and reagent-free nature of the BLYAhS reporter assay facilitated near-continuous, automated signal acquisition without additional hands-on effort and cost, providing a simple and cost-effective method for rapid DLC detection.

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Acknowledgements

Portions of this work were supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture under agreement number 2015-33610-23598 of the Small Business Innovation Research Grants Program and the University of Tennessee Institute for a Secure and Sustainable Environment (ISSE) Seed Grant.

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Authors and Affiliations

  1. Center for Environmental Biotechnology, The University of Tennessee Knoxville, 676 Dabney Hall, 1416 Circle Drive, Knoxville, TN, 37996, USA

    Tingting Xu, Anna Young & Enolia Marr

  2. 490 BioTech Inc., 2450 EJ Chapman Drive, Knoxville, TN, 37996, USA

    Gary Sayler, Steven Ripp & Dan Close

Authors
  1. Tingting Xu
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  2. Anna Young
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  3. Enolia Marr
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Correspondence to Dan Close.

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Conflict of interest

Gary Sayler, Steven Ripp, and Dan Close maintain Board of Director affiliations with 490 BioTech, Inc. Tingting Xu, Anna Young, and Enolia Marr declare no conflict of interest.

Additional information

Published in the topical collection Microbial Biosensors for Analytical Applications with guest editor Gérald Thouand.

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Xu, T., Young, A., Marr, E. et al. A rapid and reagent-free bioassay for the detection of dioxin-like compounds and other aryl hydrocarbon receptor (AhR) agonists using autobioluminescent yeast. Anal Bioanal Chem 410, 1247–1256 (2018). https://doi.org/10.1007/s00216-017-0780-1

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  • DOI: https://doi.org/10.1007/s00216-017-0780-1

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