A novel methodology independent of fermentation rate for assessment of the fructophilic character of wine yeast strains

Original Paper


The yeast Saccharomyces cerevisiae has a fundamental role in fermenting grape juice to wine. During alcoholic fermentation its catabolic activity converts sugars (which in grape juice are a near equal ratio of glucose and fructose) and other grape compounds into ethanol, carbon dioxide and sensorily important metabolites. However, S. cerevisiae typically utilises glucose and fructose with different efficiency: glucose is preferred and is consumed at a higher rate than fructose. This results in an increasing difference between the concentrations of glucose and fructose during fermentation. In this study 20 commercially available strains were investigated to determine their relative abilities to utilise glucose and fructose. Parameters measured included fermentation duration and the kinetics of utilisation of fructose when supplied as sole carbon source or in an equimolar mix with glucose. The data were then analysed using mathematical calculations in an effort to identify fermentation attributes which were indicative of overall fructose utilisation and fermentation performance. Fermentation durations ranged from 74.6 to over 150 h, with clear differences in the degree to which glucose utilisation was preferential. Given this variability we sought to gain a more holistic indication of strain performance that was independent of fermentation rate and therefore utilized the area under the curve (AUC) of fermentation of individual or combined sugars. In this way it was possible to rank the 20 strains for their ability to consume fructose relative to glucose. Moreover, it was shown that fermentations performed in media containing fructose as sole carbon source did not predict the fructophilicity of strains in wine-like conditions (equimolar mixture of glucose and fructose). This work provides important information for programs which seek to generate strains that are faster or more reliable fermenters.


Glucose Fructose Fermentation progress Strain comparison Composite trapezoid rule 



Area under the curve


  1. 1.
    Arroyo-Lopez FN, Querol A, Barrio E (2009) Application of a substrate inhibition model to estimate the effect of fructose concentration on the growth of diverse Saccharomyces cerevisiae strains. J Ind Microbiol Biotechnol 36:663–669PubMedCrossRefGoogle Scholar
  2. 2.
    Berthels NJ, Cordero Otero RR, Bauer FF, Thevelein JM, Pretorius IS (2004) Discrepancy in glucose and fructose utilisation during fermentation by Saccharomyces cerevisiae wine yeast strains. FEMS Yeast Res 4:683–689PubMedCrossRefGoogle Scholar
  3. 3.
    Berthels NJ, Cordero Otero RR, Bauer FF, Pretorius IS, Thevelein JM (2008) Correlation between glucose/fructose discrepancy and hexokinase kinetic properties in different Saccharomyces cerevisiae wine yeast strains. Appl Microbiol Biotechnol 77:1083–1091PubMedCrossRefGoogle Scholar
  4. 4.
    Bisson LF, Block DE (2002) Ethanol tolerance in Saccharomyces. Biodivers Biotechnol Wine Yeast, 85–98Google Scholar
  5. 5.
    Bisson LF, Fraenkel DG (1983) Involvement of kinases in glucose and fructose uptake by Saccharomyces cerevisiae. Proc Natl Acad Sci USA 80:1730–1734PubMedCrossRefGoogle Scholar
  6. 6.
    Boehringe-Mannheim (1989) d-glucose/d-fructose. In: Methods of biochemical analysis and food analysis. Boehringer Mannheim, pp 50–55Google Scholar
  7. 7.
    Cavazza A, Poznanski E, Trioli G (2004) Restart of fermentation of simulated stuck wines by direct inoculation of active dry yeasts. Am J Enol Vitic 55:160–167Google Scholar
  8. 8.
    Ciriacy M, Reifenberger E (1997) Hexose transport. In: Zimmermann FK, Entian KD (eds) Yeast sugar metabolism. Technimic, Lancaster, pp 45–65Google Scholar
  9. 9.
    Diderich JA, Schepper M, van Hoek P, Luttik MA, van Dijken JP, Pronk JT, Klaassen P, Boelens HF, de Mattos MJ, van Dam K, Kruckeberg AL (1999) Glucose uptake kinetics and transcription of HXT genes in chemostat cultures of Saccharomyces cerevisiae. J Biol Chem 274:15350–15359PubMedCrossRefGoogle Scholar
  10. 10.
    Diderich JA, Schuurmans JM, Van Gaalen MC, Kruckeberg AL, Van Dam K (2001) Functional analysis of the hexose transporter homologue HXT5 in Saccharomyces cerevisiae. Yeast 18:1515–1524PubMedCrossRefGoogle Scholar
  11. 11.
    Dumont A, Raynal C, Raginel F, Ortiz-Julien A (2009) The ability of wine yeast to consume fructose. Aust NZ Grapegrow Winemak 543:52–56Google Scholar
  12. 12.
    Elbing K, Larsson C, Bill RM, Albers E, Snoep JL, Boles E, Hohmann S, Gustafsson L (2004) Role of hexose transport in control of glycolytic flux in Saccharomyces cerevisiae. Appl Environ Microbiol 70:5323–5330PubMedCrossRefGoogle Scholar
  13. 13.
    Gafner J, Schütz M (1996) Impact of glucose-fructose-ratio on stuck fermentations: practical experiences to restart stuck fermentations. Wein-Wissenschaft Vitic Enol Sci 51:214–218Google Scholar
  14. 14.
    Gonçalves P, Rodrigues de Sousa H, Spencer-Martins I (2000) FSY1, a novel gene encoding a specific fructose/H+ symporter in the type strain of Saccharomyces carlsbergensis. J Bacteriol 182:5628–5630PubMedCrossRefGoogle Scholar
  15. 15.
    Guillaume C, Delobel P, Sablayrolles JM, Blondin B (2007) Molecular basis of fructose utilization by the wine yeast Saccharomyces cerevisiae: a mutated HXT3 allele enhances fructose fermentation. Appl Environ Microbiol 73:2432–2439PubMedCrossRefGoogle Scholar
  16. 16.
    Henschke PA, Jiranek V (1993) Yeast—metabolism of nitrogen compound. In: Fleet GH (ed) Wine microbiology and biotechnology. Harwood Academic, Chur, pp 77–164Google Scholar
  17. 17.
    Jiranek V, Langridge P, Henschke PA (1995) Amino-acid and ammonium utilization by Saccharomyces cerevisiae wine yeasts from a chemically-defined medium. Am J Enol Vitic 46:75–83Google Scholar
  18. 18.
    Júnior MM, Batistote M, Ernandes JR (2008) Glucose and fructose fermentation by wine yeasts in media containing structurally complex nitrogen sources. J Inst Brew 114:199–204Google Scholar
  19. 19.
    Karpel JE, Place WR, Bisson LF (2008) Analysis of the major hexose transporter genes in wine strains of Saccharomyces cerevisiae. Am J Enol Vitic 59:265–275Google Scholar
  20. 20.
    Lee CK (1987) The chemistry and biochemistry of the sweetness of sugars. Adv Carbohydr Chem Biochem 45:199–351PubMedCrossRefGoogle Scholar
  21. 21.
    Luyten K, Riou C, Blondin B (2002) The hexose transporters of Saccharomyces cerevisiae play different roles during enological fermentation. Yeast 19:713–726PubMedCrossRefGoogle Scholar
  22. 22.
    McBryde C, Gardner JM, de Barros Lopes M, Jiranek V (2006) Generation of novel wine yeast strains by adaptive evolution. Am J Enol Vitic 57:423–430Google Scholar
  23. 23.
    Meneses FJ, Henschke PA, Jiranek V (2002) A survey of industrial strains of Saccharomyces cerevisiae reveals numerous altered patterns of maltose and sucrose utilisation. J Inst Brew 108:310–321Google Scholar
  24. 24.
    Meneses FJ, Jiranek V (2002) Expression patterns of genes and enzymes involved in sugar catabolism in industrial Saccharomyces cerevisiae strains displaying novel fermentation characteristics. J Inst Brew 108:322–335Google Scholar
  25. 25.
    Ozcan S, Johnston M (1995) Three different regulatory mechanisms enable yeast hexose transporter (HXT) genes to be induced by different levels of glucose. Mol Cell Biol 15:1564–1572PubMedGoogle Scholar
  26. 26.
    Ozcan S, Johnston M (1999) Function and regulation of yeast hexose transporters. Microbiol Mol Biol Rev 63:554–569PubMedGoogle Scholar
  27. 27.
    Perez M, Luyten K, Michel R, Riou C, Blondin B (2005) Analysis of Saccharomyces cerevisiae hexose carrier expression during wine fermentation: both low- and high-affinity Hxt transporters are expressed. FEMS Yeast Res 5:351–361PubMedCrossRefGoogle Scholar
  28. 28.
    Ramos J, Szkutnicka K, Cirillo VP (1988) Relationship between low- and high-affinity glucose transport systems of Saccharomyces cerevisiae. J Bacteriol 170:5375–5377PubMedGoogle Scholar
  29. 29.
    Reifenberger E, Freidel K, Ciriacy M (1995) Identification of novel HXT genes in Saccharomyces cerevisiae reveals the impact of individual hexose transporters on glycolytic flux. Mol Microbiol 16:157–167PubMedCrossRefGoogle Scholar
  30. 30.
    Rolland F, Wanke V, Cauwenberg L, Ma PS, Boles E, Vanoni M, de Winde JH, Thevelein JM, Winderickx J (2001) The role of hexose transport and phosphorylation in cAMP signalling in the yeast Saccharomyces cerevisiae. FEMS Yeast Res 1:33–45PubMedGoogle Scholar
  31. 31.
    Rolland F, Winderickx J, Thevelein JM (2001) Glucose-sensing mechanisms in eukaryotic cells. Trends Biochem Sci 26:310–317PubMedCrossRefGoogle Scholar
  32. 32.
    Salmon J-M (1989) Effects of sugar transport inactivation in Saccharomyces cerevisiae on sluggish and stuck enological fermentations. Appl Environ Microbiol 55:953–958PubMedGoogle Scholar
  33. 33.
    Salmon JM, Vincent O, Mauricio JC, Bely M, Barre P (1993) Sugar-transport inhibition and apparent loss of activity in Saccharomyces cerevisiae as a major limiting factor of enological fermentations. Am J Enol Vitic 44:56–64Google Scholar
  34. 34.
    Serrano R, Delafuente G (1974) Regulatory properties of the constitutive hexose transport in Saccharomyces cerevisiae. Mol Cell Biochem 5:161–171PubMedCrossRefGoogle Scholar
  35. 35.
    Stanley D, Bandara A, Fraser S, Chambers PJ, Stanley GA (2010) The ethanol stress response and ethanol tolerance of Saccharomyces cerevisiae. J Appl Microbiol. (doi:  10.1111/j.1365-2672.2009.04657.x)
  36. 36.
    Tronchoni J, Gamero A, Arroyo-Lopez FN, Barrio E, Querol A (2009) Differences in the glucose and fructose consumption profiles in diverse Saccharomyces wine species and their hybrids during grape juice fermentation. Int J Food Microbiol 134:237–243PubMedCrossRefGoogle Scholar
  37. 37.
    Varela C, Cardenas J, Melo F, Agosin E (2005) Quantitative analysis of wine yeast gene expression profiles under winemaking conditions. Yeast 22:369–383PubMedCrossRefGoogle Scholar
  38. 38.
    Verwaal R, Paalman JW, Hogenkamp A, Verkleij AJ, Verrips CT, Boonstra J (2002) HXT5 expression is determined by growth rates in Saccharomyces cerevisiae. Yeast 19:1029–1038PubMedCrossRefGoogle Scholar
  39. 39.
    Wang D, Xu Y, Hu J, Zhao G (2004) Fermentation kinetics of different sugars by apple wine yeast Saccharomyces cerevisiae. J Inst Brew Distill 110:340–346Google Scholar

Copyright information

© Society for Industrial Microbiology 2010

Authors and Affiliations

  1. 1.School of Agriculture, Food and WineThe University of AdelaideGlen OsmondAustralia
  2. 2.Wine Innovation ClusterAdelaideAustralia
  3. 3.The Australian Wine Research InstituteGlen OsmondAustralia

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