Identification of novel GAPDH-derived antimicrobial peptides secreted by Saccharomyces cerevisiae and involved in wine microbial interactions
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Saccharomyces cerevisiae plays a primordial role in alcoholic fermentation and has a vast worldwide application in the production of fuel-ethanol, food and beverages. The dominance of S. cerevisiae over other microbial species during alcoholic fermentations has been traditionally ascribed to its higher ethanol tolerance. However, recent studies suggested that other phenomena, such as microbial interactions mediated by killer-like toxins, might play an important role. Here we show that S. cerevisiae secretes antimicrobial peptides (AMPs) during alcoholic fermentation that are active against a wide variety of wine-related yeasts (e.g. Dekkera bruxellensis) and bacteria (e.g. Oenococcus oeni). Mass spectrometry analyses revealed that these AMPs correspond to fragments of the S. cerevisiae glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein. The involvement of GAPDH-derived peptides in wine microbial interactions was further sustained by results obtained in mixed cultures performed with S. cerevisiae single mutants deleted in each of the GAPDH codifying genes (TDH1-3) and also with a S. cerevisiae mutant deleted in the YCA1 gene, which codifies the apoptosis-involved enzyme metacaspase. These findings are discussed in the context of wine microbial interactions, biopreservation potential and the role of GAPDH in the defence system of S. cerevisiae.
KeywordsAntimicrobial peptides Wine microbial interactions Alcoholic fermentation Biopreservation Metacaspases Glyceraldehyde-3-phosphate dehydrogenase
The present work was financed by FEDER funds through POFC-COMPETE and by national funds through Fundação para a Ciência e a Tecnologia (FCT) in the scope of project FCOMP-01-0124-FEDER-014055. M.G.A. and J.C. acknowledge the funding support from FCT (PEst-C/EQB/LA0006/2011). Patrícia Branco is the recipient of a PhD fellowship (SFRH/BD/89673/2012) funded by FCT, Portugal. We want also to thanks to Professors Isabel Sá-Correia (IST/UTL, Lisbon, Portugal), Luísa Marinho (FCUL, Lisbon, Portugal) and Paula Ludovico (ICVS/UM, Braga, Portugal) for kindly providing some mutant strains.
- Bauer FF, Pretorius IS (2000) Yeast stress response and fermentation efficiency: how to survive the making of wine—a review. S Afr J Enol Vitic 21:27–51Google Scholar
- Bisson LF (1999) Stuck and sluggish fermentations. Am J Enol Vitic 50:107–119Google Scholar
- Comitini F, De JI, Pepe L, Mannazzu I, Ciani M (2004) Pichia anomala and Kluyveromyces wickerhamii killer toxins as new tools against Dekkera/Brettanomyces spoilage yeasts. FEMS Microbiol Lett 238:235–240. doi:10.1016/j.femsle.2004.07.040
- Comitini F, Ferretti R, Clementi F, Mannuzzu I, Ciani M (2005) Interactions between Saccharomyces cerevisiae and malolactic bacteria: preliminary characterization of a yeast proteinaceous compound(s) active against Oenococcus oeni. J Appl Microbiol 99:105–111. doi:10.1111/j.1365-2672.2005.02579.x PubMedCrossRefGoogle Scholar
- Delgado ML, O’Connor JE, Azorin I, Renau-Piqueras J, Gil ML, Gozalbo D (2001) The glyceraldehyde-3-phosphate dehydrogenase polypeptides encoded by the Saccharomyces cerevisiae TDH1, TDH2 and TDH3 genes are also cell wall proteins. Microbiol 147:411–417Google Scholar
- Fleet GH, Heard GM (1993) Yeast growth during fermentation. In: Fleet GH (ed) Wine microbiology and biotechnology. Harwood Academic Publishers, Basel, Switzerland, pp 27–54Google Scholar
- Pretorius IS (2000) Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking. Yeast 16:675–729. doi:10.1002/1097-0061(20000615)16:8<675::AID-YEA585>3.0.CO;2-B PubMedCrossRefGoogle Scholar
- Wagener J, Schneider JJ, Baxmann S, Kalbacher H, Borelli C, Nuding S, Küchler R, Wehkamp J, Kaeser MD, Mailänder-Sanchez D, Braunsdorf C, Hube B, Schild L, Forssmann W-G, Korting H-C, Liepke C, Schaller M (2013) A peptide derived from the highly conserved protein GAPDH is involved in tissue protection by different antifungal strategies and epithelial immunomodulation. J Investigat Dermatol 133:144–153. doi:10.1038/jid.2012.254 CrossRefGoogle Scholar