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Reducing haziness in white wine by overexpression of Saccharomyces cerevisiae genes YOL155c and YDR055w

  • Applied Genetics and Molecular Biotechnology
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Abstract

Grape proteins aggregate in white wine to form haze. A novel method to prevent haze in wine is the use of haze protective factors (Hpfs), specific mannoproteins from Saccharomyces cerevisiae, which reduce the particle size of the aggregated proteins. Hpf1p was isolated from white wine and Hpf2p from a synthetic grape juice fermentation. Putative structural genes, YOL155c and YDR055w, for these proteins were identified from partial amino acid sequences of Hpf1p and Hpf2p, respectively. YOL155c also has a homologue, YIL169c, in S. cerevisiae. Comparison of the partial amino acid sequence of deglycosylated-Hpf2p with the deduced protein sequence of YDR055w, confirmed five of the 15 potential N-linked glycosylation sites in this sequence were occupied. Methylation analysis of the carbohydrate moieties of Hpf2p indicated that this protein contained both N- and O-linked mannose chains. Material from fermentation supernatant of deletion strains had significantly less activity than the wild type. Moreover, YOL155c and YIL169c overexpressing strains and a strain overexpressing 6xHis-tagged Hpf2p produced greater haze protective activity than the wild type strains. A storage trial demonstrated the short to midterm stability of 6xHis-tagged Hpf2p in wine.

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References

  • Anumala KR, Taylor P (1992) A comprehensive procedure for preparation of partially methylation alditol acetates from glycoprotein carbohydrates. Anal Biochem 203:101–108

    Article  Google Scholar 

  • Ausubel FM (2001) Current protocols in molecular biology. Wiley, New York

    Book  Google Scholar 

  • Ballou CE (1976) Structure and biosynthesis of the mannan component of the yeast cell envelope. In: JN Strathern, EW Jones, Broach JR (eds) Academic, Washington DC, pp 93–158

    Google Scholar 

  • Carlsson S (1993) Isolation and charaterisation of glycoproteins. In: Fukuda M, Kobata A (eds) Oxford University Press, New York, pp 1–26

    Google Scholar 

  • Caro LHP, Tettelin H, Vossen JH, Ram AFJ, Van Den Ende H, Klis FM (1997) In sicilio identification of glycosyl-phosphatidylinositol-anchored plasma-membrane and cell wall proteins of Saccharomyces cerevisiae. Yeast 13:1477–1489

    Article  CAS  Google Scholar 

  • Dupin IVS, McKinnon BM, Ryan C, Boulay M, Markides AJ, Jones GP, Williams PJ, Waters EJ (2000a) Saccharomyces cerevisiae mannoproteins that protect wine from protein haze: their release during fermentation and lees contact and a proposal for their mechanism of action. J Agric Food Chem 48:3098–3105

    Article  CAS  Google Scholar 

  • Dupin IVS, Stockdale VJ, Williams PJ, Jones GP, Markides AJ, Waters EJ (2000b) Saccharomyces cerevisiae mannoproteins that protect wine from protein haze: evaluation of extraction methods and immunolocalization. J Agric Food Chem 48:1086–1095

    Article  CAS  Google Scholar 

  • Elder JH, Alexander S (1982) Endo-b-N-acetylglucosaminodase F: endo glycosidase from Flavobacterium meningospectucum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A 79:4540–4544

    Article  CAS  Google Scholar 

  • Feldmann H (2000) Genolevures—a novel approach to ‘evolutionary genomics’. FEBS Lett 487:1–2

    Article  CAS  Google Scholar 

  • Ferreira RB, Piçarra-Pereira MA, Monteiro S, Loureiro VB, Teixeira AR (2002) The wine proteins. Trends Food Sci Technol 12:230–239

    Article  Google Scholar 

  • Gancedo J (1998) Yeast carbon catabolite repression. Microbiol Mol Biol Rev 62:334–361

    Article  CAS  Google Scholar 

  • Hauser K, Tanner W (1989) Purification of the inducible alpha-agglutinin of Saccharomyces cerevisiae and molecular cloning of the gene. FEBS Lett 255:290–294

    Article  CAS  Google Scholar 

  • Henschke P, Jiranek V (1993) Yeasts. Metabolism of nitrogen compounds. In: F GH (eds) Wine microbiology and biotechnology. Harwood Academic, Chur, Switzerland, pp 77–164

    Google Scholar 

  • Høj PB, Tattersall DB, Adams K, Pocock KF, Hayasaka Y, van Heeswijck R, Waters E (2000). The ‘haze proteins’ of wine—a summary of properties, factors affecting their accumulation in grapes, and the amount of bentonite required for their removal from wine. ASEV 50th Anniversary Meeting, Seattle, Washington, USA, American Society of Enology and Viticulture

  • Inoue H, Nojima H, Okayama H (1990) High efficiency transformation of Escherichia coli with plasmids. Gene 96:23–28

    Article  CAS  Google Scholar 

  • Längle-Rouault F, Jacobs E (1995) A method for performing precise alterations in the yeast genome using a recyclable selectable marker. Nucleic Acids Res 23:3079–8081

    Article  Google Scholar 

  • Lau E, Bacic A (1993) Capillary gas chromatography of partially methylated alditol acetates on a high polarity, cross-linked, fused silica BPX70 column. J Chromatogr A 637:100–103

    Article  CAS  Google Scholar 

  • Ledoux V, Dulau L, Dubourdieu D (1992) Interprétation de l’amélioration de la stabilité protéique des vins au cours de l’élevage sur lies. J Int Sci Vigne Vin 26:239–251

    CAS  Google Scholar 

  • McKinnon B (1996) A study of protein haze prevention in model wine solutions. Horticulture, viticulture and oenology. Adelaide, University of Adelaide, p 79

    Google Scholar 

  • Moine-Ledoux V, Dubourdieu D (1999) An invertase fragment responsible for improving the protein stability of dry white wines. J Sci Food Agric 79:537–543

    Article  CAS  Google Scholar 

  • Mumberg D, Muller R, Funk M (1994) Regulatable promoters of Saccharomyces cerevisiae: comparison of transcriptional activity and their use for heterologous expression. Nucleic Acids Res 22:5767–5768

    Article  CAS  Google Scholar 

  • Needs PW, Selvendran R (1994) A critical assessment of a one-tubed procedure for the linkage analysis of polysaccharides as partially methylated alditol acetates. Carbohydr Res 254:229–244

    Article  CAS  Google Scholar 

  • Pardo M, Monteoliva L, Pla J, Sánchez M, Gil C, Nombela C (1999) Two-dimensional analysis of proteins secreted by Saccharomyces cerevisiae regenerating protoplasts: a novel approach to study the cell wall. Yeast 15:459–472

    Article  CAS  Google Scholar 

  • Pardo M, Monteoliva L, Vazquez P, Martinez R, Molero G, Nombela C, Gil C (2004) PST1 and ECM33 encode two yeast cell surface GPI proteins important for cell wall integrity. Microbiology-SGM 150:4157–4170

    Article  CAS  Google Scholar 

  • Pellerin P, Waters E, Brillouet J-M, Moutounet M (1994) Effet de polysaccharides sur la formation de trouble proteique dans un vin blanc (Effect of polysaccharides on protein haze formation in white wine). J Int Sci Vigne Vin 28:213–225

    CAS  Google Scholar 

  • Pocock KF, Rankine BC (1973) Heat test for detecting protein instability in wine. Aust Wine Brew Spirit Rev 91(5):42–43

    Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Schultz C, Gilson P, Oxley D, Youl J, Bacic A (1998) GPI-anchors on arabinogalactanproteins: implications for signalling in plants. Trends Plant Sci 3:426–431

    Article  Google Scholar 

  • Shimoi H, Sakamoto K, Okuda M, Atthi R, Iwashita K, Ito K (2002) The AWA1 gene is required for the foam-forming phenotype and cell surface hydrophobicity of sake yeast. Appl Environ Microbiol 68:2018–2025

    Article  CAS  Google Scholar 

  • Tattersall DB, Pocock KF, Hayasaka Y, Adams K, van Heeswijck R, Waters EJ, Høj PB (2001) Pathogenesis related proteins—their accumulation in grapes during berry growth and their involvement in white wine heat instability. Current knowledge and future perspectives in relation to winemaking practices. In: Roubelakis-Angelakis KA (ed) Kluwer, Dordrecht, The Netherlands, pp 183–201

    Google Scholar 

  • Wach A, Brachat A, Alberti-Segui C, Rebiscung C, Philippsen P (1997) Heterologous HIS3 marker and GFP reporter modules for PCR-targeting in Saccharomyces cerevisiae. Yeast 13:1065–1075

    Article  CAS  Google Scholar 

  • Waters EJ, Wallace W, Williams PJ (1991) Heat haze characteristics of fractionated wine proteins. Am J Enol Vitic 42:123–127

    CAS  Google Scholar 

  • Waters EJ, Wallace W, Tate ME, Williams PJ (1993) Isolation and partial characterization of a natural haze protective factor from wine. J Agric Food Chem 41:724–730

    Article  CAS  Google Scholar 

  • Waters EJ, Pellerin P, Brillouet J-M (1994a) A Saccharomyces mannoprotein that protects wine from protein haze. Carbohydr Polym 23:185–191

    Article  CAS  Google Scholar 

  • Waters EJ, Pellerin P, Brillouet J-M (1994b) A wine arabinogalactan-protein that reduces heat-induced wine protein haze. Biosci Biotechnol Biochem 58:43–48

    Article  CAS  Google Scholar 

  • Winston F, Dollard C, Ricupero-Hovasse SL (1995) Construction of a set of convenient Saccharomyces cerevisiae strains that are isogenic to S288C. Yeast 11:53–55

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Australia’s grape growers and winemakers through their investment body, the Grape and Wine Research and Development Corporation, with matching funds from the Australian Government and by the Commonwealth Cooperative Research Centres Program.

We thank Trevor Lithgow and Peter Walsh, University of Melbourne, for the gift of the plasmid p3xHA-HIS5, Jenny Bellon and Mariola Kwiatkowski of the Australian Wine Research Institute, Zofia Felton, University of Melbourne, and Jelle Lehnstein, University of Adelaide for advice and technical assistance, and Paul Chambers and Oenone Macintyre of the Australian Wine Research Institute for commenting on the manuscript.

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Author notes

  1. Shauna L. Brown

    Present address: GeneWorks Pty. Ltd, 39 Winwood St, Thebarton, SA, 5031, Australia

  2. Vanessa J. Stockdale

    Present address: Fosters Wine Estates, Wolf Blass Winery, Sturt Highway, Nuriootpa, SA, 5355, South Australia

  3. Miguel de Barros Lopes

    Present address: School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA, 5000, Australia

  4. Patrick J. Williams

    Present address: Australian Journal of Grape and Wine Research, P.O. Box 197, Glen Osmond, Adelaide, SA, 5064, Australia

  5. Peter B. Høj

    Present address: Australian Research Council, GPO Box 2702, Canberra, ACT, 2701, Australia

Authors and Affiliations

  1. The Australian Wine Research Institute, P.O. Box 197, Glen Osmond, SA, 5064, Australia

    Shauna L. Brown, Vanessa J. Stockdale, Kenneth F. Pocock, Miguel de Barros Lopes, Patrick J. Williams, Peter B. Høj & Elizabeth J. Waters

  2. School of Agriculture and Wine, Australian Centre for Plant Functional Genomics, The University of Adelaide, Waite Campus, Adelaide, SA, 5005, Australia

    Geoffrey B. Fincher

  3. Cooperative Research Centre for Bioproducts, School of Botany, University of Melbourne, Parkville, VIC, 3010, Australia

    Filomena Pettolino & Antony Bacic

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  1. Shauna L. Brown
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  2. Vanessa J. Stockdale
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Correspondence to Elizabeth J. Waters.

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Brown, S.L., Stockdale, V.J., Pettolino, F. et al. Reducing haziness in white wine by overexpression of Saccharomyces cerevisiae genes YOL155c and YDR055w . Appl Microbiol Biotechnol 73, 1363–1376 (2007). https://doi.org/10.1007/s00253-006-0606-0

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  • DOI: https://doi.org/10.1007/s00253-006-0606-0

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