Advertisement

Applied Microbiology and Biotechnology

, Volume 99, Issue 20, pp 8597–8609 | Cite as

Designing and creating Saccharomyces interspecific hybrids for improved, industry relevant, phenotypes

  • Jennifer R. BellonEmail author
  • Fei Yang
  • Martin P. Day
  • Debra L. Inglis
  • Paul J. Chambers
Applied genetics and molecular biotechnology

Abstract

To remain competitive in increasingly overcrowded markets, yeast strain development programmes are crucial for fermentation-based food and beverage industries. In a winemaking context, there are many yeast phenotypes that stand to be improved. For example, winemakers endeavouring to produce sweet dessert wines wrestle with fermentation challenges particular to fermenting high-sugar juices, which can lead to elevated volatile acidity levels and extended fermentation times. In the current study, we used natural yeast breeding techniques to generate Saccharomyces spp. interspecific hybrids as a non-genetically modified (GM) strategy to introduce targeted improvements in important, wine-relevant traits. The hybrids were generated by mating a robust wine strain of Saccharomyces cerevisiae with a wine isolate of Saccharomyces bayanus, a species previously reported to produce wines with low concentrations of acetic acid. Two hybrids generated from the cross showed robust fermentation properties in high-sugar grape juice and produced botrytised Riesling wines with much lower concentrations of acetic acid relative to the industrial wine yeast parent. The hybrids also displayed suitability for icewine production when bench-marked against an industry standard icewine yeast, by delivering icewines with lower levels of acetic acid. Additionally, the hybrid yeast produced wines with novel aroma and flavour profiles and established that choice of yeast strain impacts on wine colour. These new hybrid yeasts display the desired targeted fermentation phenotypes from both parents, robust fermentation in high-sugar juice and the production of wines with low volatile acidity, thus establishing their suitability for wine styles that are traditionally troubled by excessive volatile acidity levels.

Keywords

Saccharomyces interspecific hybrids Targeted wine yeast strain development Non-genetically modified (non-GM) High-sugar fermentation 

Notes

Acknowledgments

This work was financially supported by Australia’s grapegrowers and winemakers through their investment body the Australian Grape and Wine Authority, with matching funds from the Australian Government. The AWRI is part of the Wine Innovation Cluster.

The icewine component of this research was financed by the Natural Sciences and Engineering Research Council of Canada.

The authors would like to thank Jean-Michel Salmon (INRA, France) for his generous gift of tetraploid yeast strain 53–7 and Nick Van Holst Pellekaan for his assistance with the fluorescence flow cytometry analyses.

Funding

This study was funded by the Australian Grape and Wine Authority (project number AWR 1301) and the Natural Sciences and Engineering Research Council of Canada (Discovery grant number 238872).

Conflict of interest

The authors state that they have no conflicts of interest to disclose.

Ethical statement

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

Supplementary material

253_2015_6737_MOESM1_ESM.pdf (1.1 mb)
ESM 1 (PDF 1175 kb)

References

  1. Ausubel F, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. Wiley, New YorkGoogle Scholar
  2. Bakker J, Bridle P, Timberlake CF (1986) Tristimulus measurements (CIELAB 76) of port wine colour. Vitis 25:67–78Google Scholar
  3. Belloch C, Orlic S, Barrio E, Querol A (2008) Fermentative stress adaptation of hybrids within the Saccharomyces sensu stricto complex. Int J Food Microbiol 122:188–195CrossRefPubMedGoogle Scholar
  4. Bellon JR, Eglinton JM, Siebert TE, Pollnitz AP, Rose L, de Barros LM, Chambers PJ (2011) Newly generated interspecific wine yeast hybrids introduce flavour and aroma diversity to wines. Appl Microbiol Biotechnol 91:603–612CrossRefPubMedGoogle Scholar
  5. Bellon JR, Schmid F, Capone DL, Chambers PJ (2013) Introducing a new breed of wine yeast: interspecific hybridization between a commercial Saccharomyces cerevisiae wine yeast and Saccharomyces mikatae. PLoS ONE 8(4):e62053. doi: 10.1371/journal.pone0062053 PubMedCentralCrossRefPubMedGoogle Scholar
  6. Blomberg A, Adler L (1989) Roles of glycerol and glycerol-3-phosphate dehydrogenase (NAD+) in acquired osmotolerance of Saccharomyces cerevisiae. J Bacteriol 171:1087–1092PubMedCentralPubMedGoogle Scholar
  7. Borneman AR, Desany BA, Riches R, Affourtit JP, Forgan AH, Pretorius IS, Egholm M, Chambers PJ (2012) The genome sequence of the wine yeast Vin7 reveals an allotriploid genome with Saccharomyces cerevisiae and Saccharomyces kudriavzevii origins. FEMS Yeast Res 12:88–96CrossRefPubMedGoogle Scholar
  8. Castellari L, Ferruzzi M, Magrini A, Giudici P, Passarelli P, Zambonelli C (1994) Unbalanced wine fermentation by cryotolerant vs. non-cryotolerant Saccharomyces strains. Vitis 33:49–52Google Scholar
  9. Cheynier V, Trousdale E, Singleton VL, Salgues M, Wdlde R (1986) Characterisation of 2-S-glutathionyl caftaric acid and its hydrolysis in relation to grape wines. J Agric Food Chem 34:217–221CrossRefGoogle Scholar
  10. Cheynier V, Rigaud J, Souquet J-M, Duprat F, Moutounet M (1990) Must browning in relation to the behavior of phenolic compounds during oxidation. Am J Enol Vitic 41:346–349Google Scholar
  11. Cliff MA, Pickering GJ (2006) Determination of odour detection thresholds for acetic acid and ethyl acetate in ice wine. J Wine Res 17:45–52CrossRefGoogle Scholar
  12. Curtin CD, Borneman AR, Chambers PC, Pretorius IS (2012) De-novo assembly and analysis of the heterozygous triploid genome of the wine spoilage yeast Dekkera bruxellensis AWRI 1499. PLoS ONE 7(3):e33840. doi: 10.1371/journal.pone0033840 PubMedCentralCrossRefPubMedGoogle Scholar
  13. Damasceno LF, Fernandes FA, Magalhães MM, Brito ES (2008) Non-enzymatic browning in clarified apple juice during thermal treatment: kinetics and process control. Food Chem 106:172–179CrossRefGoogle Scholar
  14. Eglinton JM, McWilliam SJ, Fogarty MW, Francis IL, Kwiatkowski MJ, Høj PB, Henschke PA (2000) The effect of Saccharomyces bayanus-mediated fermentation on the chemical composition and aroma profile of Chardonnay wine. Aust J Grape Wine Res 6:190–196CrossRefGoogle Scholar
  15. Esteve-Zarzoso B, Belloch C, Uruburu F, Querol A (1999) Identification of yeasts by Saccharomyces cerevisiae heterozygous for mating type. Genetics 70:41–58Google Scholar
  16. Etiévant PX (1991) Wine. In: Maarse H (ed) Volatile compounds in food. Food Science and Technology. Marcel Dekker Inc, New York, pp 362–371Google Scholar
  17. Fortuna M, Sousa MJ, Corte-Real M, Leao C, Salvador A, Sansonetty F (2001) Cell cycle analysis of yeasts. Curr Protocol Cytom 11.13.1–11.13.9Google Scholar
  18. Gafner J, Schütz M (1996) Impact of glucose-fructose-ratio on stuck fermentations; practical experiences to restart stuck fermentations. Vitic Enol Sci 51:214–218Google Scholar
  19. Greig D, Borts RH, Louis EJ, Travisano M (2002) Epistasis and hybrid sterility in Saccharomyces. Proc R Soc Lond 269:1167–1171CrossRefGoogle Scholar
  20. Kontkanen D, Inglis D, Pickering G, Reynolds A (2004) Effect of yeast inoculation rate, acclimatization, and nutrient addition on Icewine fermentation. Am J Enol Vitic 55:363–370Google Scholar
  21. Kumaran R, Yang SY, Leu JY (2013) Characterization of chromosome stability in diploid, polyploidy and hybrid yeast cells. PLoS One 8:e68094PubMedCentralCrossRefPubMedGoogle Scholar
  22. Kunicka-Styczyńska A, Rajkowska K (2011) Physiological and genetic stability of hybrids of industrial wine yeasts Saccharomyces sensu stricto complex. J Appl Microbiol 110:1538–1549CrossRefPubMedGoogle Scholar
  23. Mayer VW, Aguilera A (1990) High levels of chromosome instability in polyploids of Saccharomyces cerevisiae. Mutat Res 231:177–186CrossRefPubMedGoogle Scholar
  24. Morales L, Dujon B (2012) Evolutionary role of interspecies hybridization and genetic exchanges in yeasts. Microbiol Mol Biol Rev 76:721–739PubMedCentralCrossRefPubMedGoogle Scholar
  25. Naumov GI (1996) Genetic identification of biological species in the Saccharomyces sensu stricto complex. J Ind Microbiol 17:295–302CrossRefGoogle Scholar
  26. Nissen TL, Schulze U, Nielsen J, Villadsen J (1997) Flux distributions in anaerobic, glucose-limited cultures of Saccharomyces cerevisiae. Microbiology 143:203–218CrossRefPubMedGoogle Scholar
  27. Nurgel C, Pickering GJ, Inglis DL (2004) Sensory and chemical characteristics of Canadian ice wines. J Sci Food Agric 84:1675–1684CrossRefGoogle Scholar
  28. Pérez-Través L, Lopes CA, Querol A, Barrio E (2014) On the complexity of the Saccharomyces bayanus taxon: hybridisation and potential hybrid speciation. PLoS One 9:e93729PubMedCentralCrossRefPubMedGoogle Scholar
  29. Rainieri S, Zambonelli C, Giudici P, Castellari L (1998) Characterisation of thermotolerant Saccharomyces cerevisiae hybrids. Biotechnol Lett 20:543–547CrossRefGoogle Scholar
  30. Reifenberger E, Boles E, Ciracy M (1997) Kinetic characterization of individual hexose transporters of Saccharomyces cerevisiae and their relation to the triggering mechanisms of glucose repression. Eur J Biochem 245:324–333CrossRefPubMedGoogle Scholar
  31. Rodrigues de Sousa H, Spencer-Martins I, Gonçalves P (2004) Differential regulation by glucose and fructose of a gene encoding a specific fructose/H+ symporter in Saccharomyces sensu stricto yeasts. Yeast 21:519–530CrossRefPubMedGoogle Scholar
  32. Saison D, De Schutter DP, Uyttenhove B, Delvaux F, Delvaux FR (2009) Contribution of staling compounds to the aged flavor of lager beer by studying their flavor thresholds. Food Chem 114:1206–1215CrossRefGoogle Scholar
  33. Salgues M, Cheynier V, Gunata Z, Wylde R (1986) Oxidation of grape juice 2-S-glutathionyl caffeoyl tartaric acid by Botrytis cinerea laccase and characterization of a new substance: 2,5-di-S-glutathionyl caffeoyl tartaric acid. J Food Sci 51:1191–1194CrossRefGoogle Scholar
  34. Salmon JM (1997) Enological fermentation kinetics of an isogenic ploidy series derived from an industrial Saccharomyces cerevisiae strain. J Ferment Bioeng 83:253–260CrossRefGoogle Scholar
  35. Sebastini F, Barberio Casalone E, Cavalieri D, Polsinelli M (2002) Crosses between Saccharomyces cerevisiae and Saccharomyces bayanus generate fertile hybrids. Res Microbiol 153:53–58CrossRefGoogle Scholar
  36. Siebert TE, Smyth HE, Capone DL, Neuwöhner C, Pardon KH, Skouroumounis GK, Herderich MJ, Sefton MA, Pollnitz AP (2005) Stable isotope dilution analysis of wine fermentation products by HS-SPME-GC-MS. Anal Bioanal Chem 381:937–947CrossRefPubMedGoogle Scholar
  37. Walker GM (1998) Yeast physiology and biotechnology. Wiley, LondonGoogle Scholar
  38. Wieczorke R, Krampe S, Weierstall T, Friedel K, Hollenberg CP, Boles E (1999) Concurrent knockout of at least 20 transporter genes is required to block uptake of hexoses in Saccharomyces cerevisiae. FEBS Lett 464:123–128CrossRefPubMedGoogle Scholar
  39. Zambonelli C, Passarelli P, Rainieri S, Bertolini L (1997) Technological properties and temperature response of interspecific Saccharomyces hybrids. J Sci Food Agric 74:7–12CrossRefGoogle Scholar
  40. Zoecklein BW, Fugelsang KC, Gump BH, Nury FS (1995a) Wine analysis and production. Chapter: volatile acidity. Chapman and Hall, New York, pp 192–198CrossRefGoogle Scholar
  41. Zoecklein BW, Fugelsang KC, Gump BH, Nury FS (1995b) Wine analysis and production. Chapter: laboratory procedures. Chapman and Hall, New York, pp 474–477CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Jennifer R. Bellon
    • 1
    Email author
  • Fei Yang
    • 2
  • Martin P. Day
    • 1
  • Debra L. Inglis
    • 2
  • Paul J. Chambers
    • 1
  1. 1.The Australian Wine Research InstituteGlen OsmondAustralia
  2. 2.Cool Climate Oenology and Viticulture InstituteBrock UniversitySt CatharinesCanada

Personalised recommendations