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Indole-based assay to assess the effect of ethanol on Pseudomonas putida F1 dioxygenase activity

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Abstract

Toluene dioxygenase (TDO) is ubiquitous in nature and has a broad substrate range, including benzene, toluene, ethylbenzene and xylenes (BTEX). Pseudomonas putida F1 (PpF1) induced on toluene is known to produce indigo from indole through the activity of TDO. In this work, a spectrophotometric assay previously developed to measure indole to indigo production rates was modified to characterize the effects of various ethanol concentrations on toluene aerobic biodegradation activity and assess catabolite repression of TDO. Indigo production rate by cells induced on toluene alone was 0.0012 ± 0.0006 OD610 min−1. The presence of ethanol did not fully repress TDO activity when toluene was also available as a carbon source. However, indigo production rates by PpF1 grown on ethanol:toluene mixtures (3:1 w/w) decreased by approximately 50%. Overall, the proposed spectrophotometric assay is a simple approach to quantify TDO activity, and demonstrates how the presence of ethanol in groundwater contaminated with reformulated gasoline is likely to interfere with naturally occurring microorganisms from fully expressing their aerobic catabolic potential towards hydrocarbons bioremediation.

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Authors and Affiliations

  1. Tecnoamb Tecnologias Ambientais Ltda, Desembargador Vitor Lima, 260 Sala 604, Florianópolis, SC, 88040-400, Brazil

    Márcio Luis Busi da Silva

  2. Department of Civil and Environmental Engineering, Rice University, MS 317, Houston, TX, 77251-1892, USA

    Pedro J. J. Alvarez

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  1. Márcio Luis Busi da Silva
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  2. Pedro J. J. Alvarez
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Correspondence to Márcio Luis Busi da Silva.

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da Silva, M.L.B., Alvarez, P.J.J. Indole-based assay to assess the effect of ethanol on Pseudomonas putida F1 dioxygenase activity. Biodegradation 21, 425–430 (2010). https://doi.org/10.1007/s10532-009-9312-9

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  • DOI: https://doi.org/10.1007/s10532-009-9312-9

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