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Biocontrol of Brettanomyces/Dekkera bruxellensis in alcoholic fermentations using saccharomycin-overproducing Saccharomyces cerevisiae strains

  • Patrícia Branco
  • Farzana Sabir
  • Mário Diniz
  • Luísa Carvalho
  • Helena AlbergariaEmail author
  • Catarina PristaEmail author
Applied genetics and molecular biotechnology
  • 33 Downloads

Abstract

Microbial contamination of alcoholic fermentation processes (e.g. winemaking and fuel-ethanol production) is a serious problem for the industry since it may render the product unacceptable and/or reduce its productivity, leading to large economic losses. Brettanomyces/Dekkera bruxellensis is one of the most dangerous microbial contaminant of ethanol industrial fermentations. In the case of wine, this yeast species can produce phenolic compounds that confer off-flavours to the final product. In fuel-ethanol fermentations, D. bruxellensis is a persistent contaminant that affects ethanol yields and productivities. We recently found that Saccharomyces cerevisiae secretes a biocide, which we named saccharomycin, composed of antimicrobial peptides (AMPs) derived from the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Saccharomycin is active against several wine-related yeast species, namely D. bruxellensis. However, the levels of saccharomycin naturally secreted by S. cerevisiae during alcoholic fermentation are not sufficient to ensure the complete death of D. bruxellensis. Therefore, the aim of the present work was to construct genetically modified S. cerevisiae strains to overproduce these GAPDH-derived AMPs. The expression levels of the nucleotides sequences encoding the AMPs were evaluated in the modified S. cerevisiae strains by RT-qPCR, confirming the success of the recombinant approach. Furthermore, we confirmed by immunological tests that the modified S. cerevisiae strains secreted higher amounts of the AMPs by comparison with the non-modified strain, inducing total death of D. bruxellensis during alcoholic fermentations.

Keywords

Antimicrobial peptides, glyceraldehyde 3-phosphate dehydrogenase Microbial contamination Genetically-modified yeasts Wine Preservatives Fuel-ethanol Bioethanol 

Notes

Acknowledgements

The authors thank Fundação para a Ciência e Tecnologia for financial support of this work through the project PTDC/BII-BIO/31761/2017 and also through the Research Units “Linking Landscape, Environment, Agriculture and Food” (LEAF) and “Unidade de Ciências Biomoleculares Aplicadas” (UCIBIO). P. Branco was the recipient of a PhD fellowship (SFRH/ BD/ 89673/ 2012) funded by FCT, Portugal.

Funding

The present work was financed by FEDER funds through POFC-COMPETE in the scope of project FCOMP-01-0124-FEDER-014055 and by national funds through Fundação para a Ciência e Tecnologia (FCT) in the scope of project PTDC/BII-BIO/31761/2017 and also through the research units LEAF (UID/AGR/04129/2013) and UCIBIO (UID/Multi/04378/2013). P. Branco received a PhD fellowship (SFRH/ BD/ 89673/ 2012) funded by FCT, Portugal.

Compliance with ethical standards

Conflict of interest

The authors declare they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Linking Landscape, Environment, Agriculture and Food (LEAF), Instituto Superior de AgronomiaUniversity of LisbonLisbonPortugal
  2. 2.Unit of Bioenergy, Laboratório Nacional de Energia e Geologia (LNEG)LisbonPortugal
  3. 3.Research Institute for Medicines (iMed.ULisboa), Faculty of PharmacyUniversidade de LisboaLisbonPortugal
  4. 4.UCIBIO-REQUIMTE, Department of Chemistry, Faculdade de Ciências e TecnologiaUniversidade NOVA de LisboaCaparicaPortugal

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