Modification by glucose of the flocculent phenotype of a Kloeckera apiculata wine strain

  • Oscar A. Sosa
  • María C. Manca de Nadra
  • Marta E. Farías
Original Paper

Abstract

We have evaluated the induction of the flocculent phenotype of Kloeckera apiculata by glucose mc1 and propose a pathway involved in carbohydrate flocculation induction. Pulses of glucose were given to cells growing in glucose-poor medium (2 g l−1) and the flocculation percentage was measured. To elucidate the mechanism involved in flocculation induction, cycloheximide was injected into the cultures 120 min before the glucose pulse. 2,4-Dinitrophenol or cAMP was added to the media instead, or simultaneously with glucose, while a protein kinase A (PKA) inhibitor was added 30 min before the glucose pulse. With 20 and 50 g l−1 glucose pulse, the yeast flocculation percentage arises to 55 and 65%, respectively. The quantity of proteins and the reflocculating capacity of a lectinic protein extract from the yeast cell wall increase as the concentration of glucose pulse was higher. Cycloheximide prevented the glucose-induced flocculation, while cAMP or 2,4-dinitrophenol increased it 4- and 5-fold, respectively. PKA inhibitor completely prevented the glucose induction flocculation. The flocculent phenotype of K. apiculata mc1 was induced by glucose and the mechanism seems to imply de novo protein (lectin) synthesis via the PKA transduction pathway. This work contributes to the elucidation of the mechanism involved in flocculation induction by glucose of a non-Saccharomyces wine yeast, K. apiculata, which has not been reported. The induction of flocculation by glucose could be a biotechnological tool for the early removal of the indigenous microorganisms from the grape must before the inoculation of a selected starter strain to conduct the alcohol fermentation.

Keywords

Kloeckera apiculata Flocculation Flocculent phenotype Glucose induction PKA pathway cAMP effect Wine yeasts 

Notes

Acknowledgments

This work was supported by grants from Banco Río, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) and Consejo de Investigaciones de la Universidad Nacional de Tucumán (CIUNT), Argentina.

References

  1. 1.
    Al-Mahmood C, Colin S, Bonaly R (1991) Kluyveromyces bulgaricus yeast lectins: isolation of two galactose specific lectins forms from yeast cell wall. J Biol Chem 266:20882–20887Google Scholar
  2. 2.
    Amri MA, Bonaly R, Duteurtre B, Moll M (1979) Growth and flocculation of two Saccharomyces uvarum strain. Eur J Appl Microbiol Biotechnol 7:227–234CrossRefGoogle Scholar
  3. 3.
    Bendiak DS (1994) Quantification of the Helm’s flocculation test. J Am Soc Brew Chem 52:120–122Google Scholar
  4. 4.
    Caspani G, Tortora P, Hanozet GM, Guerritore A (1985) Glucose-stimulated cAMP increase may be mediated by intracellular acidification in Saccharomyces cerevisiae. FEBS Lett 186:75–79CrossRefGoogle Scholar
  5. 5.
    Crauwels M, Donaton MCV, Pernambuca MB, Winderickx J, de Winde JH, Thevelein JM (1997) The Sch9 protein kinase in the yeast Saccharomyces cerevisiae control cAPK activity and is required for nitrogen activation of the fermentable-growth-medium induced (FMG)-pathway. Microbiology 143:2627–2637CrossRefGoogle Scholar
  6. 6.
    El-Behhari M, Ngondi Ekomé J, Pucci B, Coulon J, Bonaly R (1998) Comparative extraction procedures for a galactose specific lectin involved in flocculation of Kluyveromyces lactis strains. Appl Microbiol Biotechnol 49:16–23CrossRefGoogle Scholar
  7. 7.
    Farías ME, Manca de Nadra MC (2003) Flocculation and cell surface characterization of Kloeckera apiculata form wine. J Appl Microbiol 95:457–462CrossRefGoogle Scholar
  8. 8.
    Fernandes PA, Sousa M, Morales-Ferreira P (1993) Flocculation of Kluyveromyces marxianus is induced by temperature upshift. Yeast 9:859–869CrossRefGoogle Scholar
  9. 9.
    Gagiano M, Bauer FF, Pretorius IS (2002) The sensing of nutritional status and the relationship to filamentous growth in Saccharomyces cerevisiae. FEMS Yeast Res 2:433–470Google Scholar
  10. 10.
    Gagiano M, Bester M, van Dyk D, Franken J, Bauer FF, Pretorius IS (2003) Mss11p is a transcription factor regulating pseudohyphal differentiation, invasive growth and starch metabolism in Saccharomyces cerevisiae in response to nutrient availability. Mol Microbiol 47:119–134CrossRefGoogle Scholar
  11. 11.
    Géhin G, Bonaly R, Coulon J (2001) The role of glucose in the Kluyveromyces bulgaricus flocculation phenomenon: transduction by cAMP-dependent protein kinase pathway? FEMS Microbiol Lett 203:229–233CrossRefGoogle Scholar
  12. 12.
    Helm E, Nohr B, Thorne RSW (1953) The measurement of yeast flocculence and its significance in brewing. Wallerstein Lab Communs 16:315–355Google Scholar
  13. 13.
    Javadekar VS, Sivaraman H, Sainkar R, Khan MI (2000) A mannose-binding protein from the cell surface of flocculent Saccharomyces cereivsiae (NCIM 3528): its role in flocculation. Yeast 16:99–110CrossRefGoogle Scholar
  14. 14.
    Jin YL, Speers RA (1999) Flocculation of Saccharomyces cerevisiae. Food Res Intern 31:421–440CrossRefGoogle Scholar
  15. 15.
    Jin YL, Speers RA (2000) Effect of environmental conditions on the flocculation of Saccharomyces cerevisiae. J Am Soc Brew Chem 58:108–116Google Scholar
  16. 16.
    Johnson BF, Walker T, Calleja GB, Seligy VL (1988) Sexual co-flocculation and asexual self-flocculation in budding and fission yeast: experimental establishment of a fundamental difference. Can J Microbiol 34:1105–1107CrossRefGoogle Scholar
  17. 17.
    Kim TS, Kim HY, Yoon JH, Kang HS (2004) Recruitment of the Swi/Snf complex by Ste12-Tec1 promotes Flo8-Mss11-mediated activation of STA1 expression. Mol Cell Biol 24:9542–9556CrossRefGoogle Scholar
  18. 18.
    Kraakman L, Lemaire K, Ma P, Teunissen WRH, Donaton MCV, van Dijck P, Winderickx J, de Winde J, Thevelein JM (1999) A Saccharomyces cerevisiae G-protein coupled receptor Gpr1, is specifically required for glucose activation of the cAMP pathway during transition to growth on glucose. Mol Microbiol 32:1002–1012CrossRefGoogle Scholar
  19. 19.
    Madhani H, Fink G (1997) Combinatorial control required for the specificity of yeast MAPK signaling. Science 275:1314–1317CrossRefGoogle Scholar
  20. 20.
    Miki BLA, Poon NH, James AP, Seligy VL (1982) Possible mechanism for flocculation interactions governed by gene FLO1 in Saccharomyces cerevisiae. J Bacteriol 50:878–889Google Scholar
  21. 21.
    Purwin C, Nicolay K, Scheffers WA, Holzer H (1986) Mechanism of control of adenylate cyclase activity in yeast by fermentable sugars and carbonyl cyanide m-chlorophenylhydrazone. J Biol Chem 261:8744–8749Google Scholar
  22. 22.
    Romano P, Fiore C, Paraggio M, Caruso M, Capece A (2003) Function of yeast species and strains in wine flavour. Int J Food Microbiol 86:169–180CrossRefGoogle Scholar
  23. 23.
    Rupp S, Summers E, Lo HJ, Madhani H, Fink G (1999) MAP kinase and cAMP filamentation signaling pathways converge on the unusually large promoter of the yeast FLO11 gene. EMBO J 18:1257–1269CrossRefGoogle Scholar
  24. 24.
    Sampermans S, Mortier J, Soares EV (2005) Flocculation onset in Saccharomyces cerevisiae: the role of nutrients. J Appl Microbiol 98:525–531CrossRefGoogle Scholar
  25. 25.
    Schwartz MA, Madhani HD (2004) Principles of MAP kinase signaling specificity in Saccharomyces cerevisiae. Annu Rev Genet 38:725–748CrossRefGoogle Scholar
  26. 26.
    Soares EV, Texeira JA, Mota M (1994) Effect of cultural and nutritional conditions on the control of flocculation expression in Saccharomyces cerevisiae. Can J Microbiol 40:851–857Google Scholar
  27. 27.
    Soares EV, Mota M (1996) Flocculation onset, growth phase, and genealogical age in Saccharomyces cerevisiae. Can J Microbiol 42:539–547CrossRefGoogle Scholar
  28. 28.
    Soares EV, Vroman A (2003) Effect of different starvation conditions on the flocculation of Saccharomyces cerevisiae. J Appl Microbiol 95:325–330CrossRefGoogle Scholar
  29. 29.
    Stewart GG (1975) Yeast flocculation. Practical implications and experimental findings. Brew Dig 50:42–62Google Scholar
  30. 30.
    Stratford M (1992) Lectin-mediated aggregation of yeasts–yeast flocculation. Biotechnol Genet Eng Rev 10:283–341Google Scholar
  31. 31.
    Stratford M (1994) Genetic aspects of yeast flocculation: in particular, the role of FLO genes in the flocculation of Saccharomyces cerevisiae. Colloids Surf B Biointerfaces 2:151–158CrossRefGoogle Scholar
  32. 32.
    Stratford M, Carter AT (1993) Yeast flocculation: lectin synthesis and activation. Yeast 9:371–378CrossRefGoogle Scholar
  33. 33.
    Straver MH, Smit G, Kijne J W (1994) Purification and partial characterization of a flocculin from brewer’s yeast. Appl Environ Microbiol 60:2754–2758Google Scholar
  34. 34.
    Teunissen AWRH, Steensma HY (1995) Review: the dominant flocculation genes of Saccharomyces cerevisiae constitute a new subtelomeric gene family. Yeast 11:1001–1013CrossRefGoogle Scholar
  35. 35.
    Teunissen AWRH, Van Der Berg J, Steensma HY (1995) Transcriptional regulation of flocculation genes in Saccharomyces cerevisiae. Yeast 11:435–446CrossRefGoogle Scholar
  36. 36.
    Thevelein JM, Cauwenberg L, Colombo S, de Winde JH, Donation M, Dumortier F, Kraakman L, Lemaire K, Ma P, Nauwealaers D, Rolland F, Teunissen A, van Dijck P, Versele M, Wera S, Winderickx J (2000) Nutrient-induced signal transduction through the protein kinase A pathway and its role in the control of metabolism, stress resistance, and growth in yeast. Enzyme Microb Technol 26:819–825CrossRefGoogle Scholar
  37. 37.
    van Dyk D, Pretorius IS, Bauer FF (2005) Mss11p is a central element of the regulatory network that controls FLO11 expression and invasive growth in Saccharomyces cerevisiae. Genetics 169:91–106Google Scholar
  38. 38.
    Verstrepen KJ, Iserentant D, Malcorps P, Derdelinckx G, van Dijck P, Winderickx J, Pretorius IS, Thevelein JM, Delvaux FR (2004) Glucose and sucrose: hazardous fast-food for industrial yeast? Trends Biotechnol 22:531–537CrossRefGoogle Scholar
  39. 39.
    Verstrepen KJ, Klis FM (2006) Flocculation, adhesion and biofilm formation in yeasts. Mol Microbiol 60:5–15CrossRefGoogle Scholar
  40. 40.
    Vyas VK, Kuchin S, Berkey CD, Carlson M (2003) Snf1 kinases with different beta-subunit isoforms play distinct roles in regulating haploid invasive growth. Mol Cell Biol 23:1341–1348CrossRefGoogle Scholar
  41. 41.
    Winderickx J, Holsbeeks I, Lagatie O, Giots F, Thevelein J, de Winde H (2003) From feast to famine: adaptation to nutrient availability in yeast. In: Hohmann S, Mager PWH (eds) Topics in current genetics, vol. 1: yeast stress responses. Springer, Berlin, pp 305–386CrossRefGoogle Scholar
  42. 42.
    Yaguchi S, Mitsui K, Iba H, Tsurugi K (2000) Phosphorylation of the GTS1 gene product of the yeast Saccharomyces cerevisiae and its effect on heat tolerance and flocculation. FEMS Microbiol Lett 187:179–189CrossRefGoogle Scholar

Copyright information

© Society for Industrial Microbiology 2008

Authors and Affiliations

  • Oscar A. Sosa
    • 1
  • María C. Manca de Nadra
    • 1
    • 2
  • Marta E. Farías
    • 1
    • 2
  1. 1.Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán and Centro de Referencia para Lactobacilos (CERELA)TucumánArgentina
  2. 2.Career Investigator of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)TucumánArgentina

Personalised recommendations