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A new method for the detection of early contamination of red wine by Brettanomyces bruxellensis using Pseudomonas putida 4-ethylphenol methylene hydroxylase (4-EPMH)

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Abstract

Brettanomyces/Dekkera bruxellensis is a cause of major concern for the winemaking industry worldwide. If a slight presence of this spoilage yeast in red wine adds a Brett character, a strong contamination has irreversible and detrimental effects on the organoleptic qualities due to the production of volatile phenols such as 4-ethylphenol. Time is a key factor in the treatment of B. bruxellensis contaminations. Nowadays, the diagnostic and quantification resources available are time consuming and too expensive, making them either inadequate or inaccessible to most of the winemakers. This study was focused on a new, easy to use, inexpensive method that could allow winemakers to directly detect B. bruxellensis contamination in red wine at an early stage, hence, reducing wine spoilage. In this work, the ability of Pseudomonas putida 4-ethylphenol methylene hydroxylase was tested in order to catabolize the 4-ethylphenol and to elaborate an enzymatic assay with the purpose of detecting early contaminations by B. bruxellensis in red wine. We have developed a colorimetric enzymatic assay, based on the redox state of the 4-ethylphenol methylene hydroxylase co-factor, cytochrome C, that can detect and quantify low concentrations of 4-ethylphenol. The range of concentrations detected is well below the level detectable by the human nose. Combined to an enrichment step, this method allows the detection of B. bruxellensis at an initial concentration of less than 10 cells per ml.

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Acknowledgements

We would like to thank Dr. David HOPPER form the University of Aberystwyth (Wales UK) for the purified 4-EPMH and his precious knowledge and advice.

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    Authors and Affiliations

    1. BIOSENTEC, 65 Allées Campferran, 31320, Auzeville-Tolosane, France

      Hélène Daniels-Treffandier & Christine Campbell

    2. Oxford Vaccine Group, CCVTM, Churchill Hospital, University of Oxford, Oxford, OX3 7LE, UK

      Hélène Daniels-Treffandier

    3. Decanter, Blue Fin Building, 110 Southwark Street, London, SE1 0SU, UK

      Christine Campbell

    4. Chemical & Petrochemical Engineering Department, Faculty of Engineering, Lebanese University, Tripoli, Lebanon

      Joyce Kheir

    5. Faculté des Sciences, Université Saint-Joseph, Mar Roukoz, Mkallès, P.O. Box 11514, Beirut, 1107-2050, Lebanon

      Dominique Salameh & Roger Lteif

    6. Laboratoire de Génie Chimique UMR CNRS 5503, INPT-ENSIACET, Université de Toulouse, 4 allée Emile Monso, BP 74233, 31432, Toulouse Cedex 04, France

      Cédric Brandam & Patricia Taillandier

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    1. Hélène Daniels-Treffandier
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    Correspondence to Joyce Kheir.

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    Hélène Daniels-Treffandier and Christine Campbell have been contributed equally to this work

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    Daniels-Treffandier, H., Campbell, C., Kheir, J. et al. A new method for the detection of early contamination of red wine by Brettanomyces bruxellensis using Pseudomonas putida 4-ethylphenol methylene hydroxylase (4-EPMH). Eur Food Res Technol 243, 1117–1125 (2017). https://doi.org/10.1007/s00217-016-2822-x

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