Saccharomyces cerevisiae accumulates GAPDH-derived peptides on its cell surface that induce death of non-Saccharomyces yeasts by cell-to-cell contact.

During wine fermentations, Saccharomyces cerevisiae starts to excrete antimicrobial peptides (AMPs) into the growth medium that induce death of non-Saccharomyces yeasts at the end of exponential growth phase (24-48 h). Those AMPs were found to derive from the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). On the other hand, the early death of non-Saccharomyces yeasts during wine fermentations was also found to be mediated by a cell-to-cell contact mechanism. Since GAPDH is a cell-wall-associated protein in S. cerevisiae, we put forward the hypothesis that the GAPDH-derived AMPs could accumulate on the cell surface of S. cerevisiae, thus inducing death of non-Saccharomyces yeasts by cell-to-cell contact. Here we show that 48-h grown (stationary phase) cells of S. cerevisiae induce death of Hanseniaspora guilliermondii and Lachancea thermotolerans by direct cell-to-cell contact, while 12-h grown cells (mid-exponential phase) do not. Immunological tests performed with a specific polyclonal antibody against the GAPDH-derived AMPs revealed their presence in the cell wall of S. cerevisiae cells grown for 48 h, but not for 12 h. Taken together, our data show that accumulation of GAPDH-derived AMPs on the cell surface of S. cerevisiae is one of the factors underlying death of non-Saccharomyces yeasts by cell-to-cell contact.

[1]  I. S. Pretorius,et al.  Yeast Stress Response and Fermentation Efficiency: How to Survive the Making of Wine - A Review , 2019, South African Journal of Enology & Viticulture.

[2]  N. Arneborg,et al.  Dominance of Saccharomyces cerevisiae in alcoholic fermentation processes: role of physiological fitness and microbial interactions , 2016, Applied Microbiology and Biotechnology.

[3]  C. Prista,et al.  Antimicrobial properties and death-inducing mechanisms of saccharomycin, a biocide secreted by Saccharomyces cerevisiae , 2016, Applied Microbiology and Biotechnology.

[4]  Patrícia Branco,et al.  Cell-to-cell contact and antimicrobial peptides play a combined role in the death of Lachanchea thermotolerans during mixed-culture alcoholic fermentation with Saccharomyces cerevisiae. , 2015, FEMS microbiology letters.

[5]  Amy C. Kelly,et al.  Saccharomyces cerevisiae , 2013, Prion.

[6]  M. Bely,et al.  An innovative tool reveals interaction mechanisms among yeast populations under oenological conditions , 2013, Applied Microbiology and Biotechnology.

[7]  Patrícia Branco,et al.  Identification of novel GAPDH-derived antimicrobial peptides secreted by Saccharomyces cerevisiae and involved in wine microbial interactions , 2013, Applied Microbiology and Biotechnology.

[8]  Chen Kai-hu Application of fluorescence in situ hybridization , 2012 .

[9]  S. Ohlmeier,et al.  Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a specific substrate of yeast metacaspase. , 2011, Biochimica et biophysica acta.

[10]  M. Sirover On the functional diversity of glyceraldehyde-3-phosphate dehydrogenase: biochemical mechanisms and regulatory control. , 2011, Biochimica et biophysica acta.

[11]  J. McGarvey,et al.  A rapid method to improve protein detection by indirect ELISA. , 2011, Biochemical and biophysical research communications.

[12]  F. Girio,et al.  Saccharomyces cerevisiae CCMI 885 secretes peptides that inhibit the growth of some non-Saccharomyces wine-related strains , 2010, Applied Microbiology and Biotechnology.

[13]  A. Sawa,et al.  Glyceraldehyde-3-phosphate Dehydrogenase Aggregate Formation Participates in Oxidative Stress-induced Cell Death* , 2009, The Journal of Biological Chemistry.

[14]  H. Santos,et al.  Improving sample treatment for in-solution protein identification by peptide mass fingerprint using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. , 2007, Journal of proteome research.

[15]  F. Girio,et al.  Cellular death of two non-Saccharomyces wine-related yeasts during mixed fermentations with Saccharomyces cerevisiae. , 2006, International journal of food microbiology.

[16]  M. Sirover New nuclear functions of the glycolytic protein, glyceraldehyde‐3‐phosphate dehydrogenase, in mammalian cells , 2005, Journal of cellular biochemistry.

[17]  D. Gozalbo,et al.  Starvation and temperature upshift cause an increase in the enzymatically active cell wall-associated glyceraldehyde-3-phosphate dehydrogenase protein in yeast. , 2003, FEMS yeast research.

[18]  N. Arneborg,et al.  Characterization of early deaths of non-Saccharomyces yeasts in mixed cultures with Saccharomyces cerevisiae , 2003, Archives of Microbiology.

[19]  N. Arneborg,et al.  Viable Saccharomyces cerevisiae cells at high concentrations cause early growth arrest of non‐Saccharomyces yeasts in mixed cultures by a cell–cell contact‐mediated mechanism , 2003, Yeast.

[20]  J. O'connor,et al.  The glyceraldehyde-3-phosphate dehydrogenase polypeptides encoded by the Saccharomyces cerevisiae TDH1, TDH2 and TDH3 genes are also cell wall proteins. , 2001, Microbiology.

[21]  Linda F. Bisson,et al.  Stuck and Sluggish Fermentations , 1999, American Journal of Enology and Viticulture.

[22]  W. Chaffin,et al.  Cell Wall and Secreted Proteins ofCandida albicans: Identification, Function, and Expression , 1998, Microbiology and Molecular Biology Reviews.

[23]  J. O'connor,et al.  The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase of Candida albicans is a surface antigen , 1997, Journal of bacteriology.

[24]  J. Findlay,et al.  A protein homologous to glyceraldehyde-3-phosphate dehydrogenase is induced in the cell wall of a flocculent Kluyveromyces marxianus. , 1992, Biochimica et biophysica acta.

[25]  V. Fischetti,et al.  A major surface protein on group A streptococci is a glyceraldehyde-3- phosphate-dehydrogenase with multiple binding activity , 1992, The Journal of experimental medicine.

[26]  E. Postma,et al.  Proton-motive force-driven D-galactose transport in plasma membrane vesicles from the yeast Kluyveromyces marxianus. , 1991, The Journal of biological chemistry.

[27]  M. Holland,et al.  Isolation and characterization of yeast strains carrying mutations in the glyceraldehyde-3-phosphate dehydrogenase genes. , 1985, The Journal of biological chemistry.

[28]  R. Dernick,et al.  Simplified method for silver staining of proteins in polyacrylamide gels and the mechanism of silver staining , 1985 .