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Microbial genotoxicity bioreporters based on sulA activation

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Abstract

A bacterial genotoxicity reporter strain was constructed in which the tightly controlled strong promoter of the Escherichia coli SOS response gene sulA was fused to the alkaline phosphatase-coding phoA reporter gene. The bioreporter responded in a dose-dependent manner to three model DNA-damaging agents—hydrogen peroxide, nalidixic acid (NA), and mitomycin C (MMC)—detected 30–60 min after exposure. Detection thresholds were 0.15 μM for MMC, 7.5 μM for nalidixic acid, and approximately 50 μM for hydrogen peroxide. A similar response to NA was observed when the bioreporter was integrated into a specially designed, portable electrochemical detection platform. Reporter sensitivity was further enhanced by single and double knockout mutations that enhanced cell membrane permeability (rfaE) and inhibited DNA damage repair mechanisms (umuD, uvrA). The rfaE mutants displayed a five- and tenfold increase in sensitivity to MMC and NA, respectively, while the uvrA mutation was advantageous in the detection of hydrogen peroxide. A similar sensitivity was displayed by the double rfaE/uvrA mutant when challenged with the pre-genotoxic agents 2-amino-3-methylimidazo[4,5-f]quinoline and 2-aminoanthracene following metabolic activation with an S9 mammalian liver fraction.

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Acknowledgments

The authors are grateful for the funding provided by the German BMBF and the Israeli MOST in the framework of project WT601 (“Dipchip”) of the German/Israeli binational Water Technology Program. We are grateful to the National BioResource Project, Japan, for generously sharing the E. coli Keio mutants’ collection.

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Authors and Affiliations

  1. Department of Plant and Environmental Sciences, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel

    Alva Biran, Sharon Yagur-Kroll, Rami Pedahzur & Shimshon Belkin

  2. Department of Physical Electronics, Faculty of Engineering, Tel Aviv University, Tel Aviv, 69978, Israel

    Hadar Ben Yoav & Yosi Shacham-Diamand

  3. Division of Qualitative Hydrology, German Federal Institute of Hydrology (BfG), Am Mainzer Tor 1, 56068, Koblenz, Germany

    Sebastian Buchinger & Georg Reifferscheid

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  1. Alva Biran
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  2. Hadar Ben Yoav
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Correspondence to Shimshon Belkin.

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Biran, A., Ben Yoav, H., Yagur-Kroll, S. et al. Microbial genotoxicity bioreporters based on sulA activation. Anal Bioanal Chem 400, 3013–3024 (2011). https://doi.org/10.1007/s00216-011-5007-2

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  • DOI: https://doi.org/10.1007/s00216-011-5007-2

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