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Anal Bioanal Chem. 2018 Feb;410(4):1247-1256. doi: 10.1007/s00216-017-0780-1. Epub 2017 Dec 6.

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

Author information

1
Center for Environmental Biotechnology, The University of Tennessee Knoxville, 676 Dabney Hall, 1416 Circle Drive, Knoxville, TN, 37996, USA.
2
490 BioTech Inc., 2450 EJ Chapman Drive, Knoxville, TN, 37996, USA.
3
490 BioTech Inc., 2450 EJ Chapman Drive, Knoxville, TN, 37996, USA. dan.close@490biotech.com.

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.

KEYWORDS:

Aryl hydrocarbon receptor; Autobioluminescence; Bioassays; Bioreporter; Dioxin-like compound; Dioxins/furans

PMID:
29214529
DOI:
10.1007/s00216-017-0780-1
[Indexed for MEDLINE]

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