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Towards an effective biosensor for monitoring AD leachate: a knockout E. coli mutant that cannot catabolise lactate

  • Applied microbial and cell physiology
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Abstract

Development of a biosensor for the convenient measurement of acetate and propionate concentrations in a two-phase anaerobic digestor (AD) requires a bacterium that will be unresponsive to the other organic acids present in the leachate, of which lactate is the most abundant. Successive gene knockouts of E.coli W3110 d-lactate dehydrogenase (dld), l-lactate dehydrogenase (lldD), glycolate oxidase (glcD) and a suspected l-lactate dehdrogenase (ykgF) were performed. The resulting quadruple mutant (IMD Wldgy) was incapable of growth on d- and l-lactate, whereas the wild type grew readily on these substrates. Furthermore, the O2 consumption rates of acetate-grown IMD Wldgy cell suspensions supplied with either acetate (0.1 mM) or a synthetic leachate including acetate (0.1 mM) and dl-lactate (1 mM) were identical (2.79 and 2.70 mg l−1 min−1, respectively). This was in marked contrast to similar experiments with the wild type which gave initial O2 consumption rates of 2.00, 2.36 and 2.97 mg l−1 min−1 when cell suspensions were supplied with acetate (0.1 mM), acetate (0.1 mM) plus d-lactate (1 mM) or acetate (0.1 mM) plus l-lactate (1 mM), respectively. The knockout strain provides a platform for the design of a biosensor that can accessibly monitor acetate and propionate concentrations in AD leachate via O2-uptake measurements.

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Acknowledgments

This publication has emanated from research conducted with the financial support of Science Foundation Ireland/Charles Parsons Energy Research Programme. The authors thank Joseph O’Neill for the help in setting up the DO probe.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. UCD Schools of Biosystems Engineering, Food and Agriculture, University College Dublin, Belfield, Dublin 4, Ireland

    Joseph Sweeney & Kevin McDonnell

  2. UCD School of Biomolecular and Biomedical Science, University College Dublin, Belfield, Dublin 4, Ireland

    Joseph Sweeney & Cormac D. Murphy

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  1. Joseph Sweeney
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  2. Cormac D. Murphy
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  3. Kevin McDonnell
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Correspondence to Cormac D. Murphy.

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Sweeney, J., Murphy, C.D. & McDonnell, K. Towards an effective biosensor for monitoring AD leachate: a knockout E. coli mutant that cannot catabolise lactate. Appl Microbiol Biotechnol 99, 10209–10214 (2015). https://doi.org/10.1007/s00253-015-6887-4

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  • DOI: https://doi.org/10.1007/s00253-015-6887-4

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