Skip to main content
SpringerLink
Log in

A quick screening method to identify β-glucosidase activity in native wine yeast strains: application of Esculin Glycerol Agar (EGA) medium

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

One hundred and fifty-four yeast strains were isolated from grapes and musts of Uruguayan vineyards and wineries. Only thirty strains showed β-glucosidase activity in Esculin Glycerol Agar (EGA) solid medium. Twenty-one were non-Saccharomyces and nine were Saccharomyces cerevisiae strains. The objective of this study was to evaluate the suitability of Esculin Glycerol Agar (EGA) solid medium for screening β-glucosidase activity in native yeasts strains. Halo sizes measured in the EGA solid medium were correlated to the Glycosyl-Glucose (GG) indexes measured after fermentation of grape musts with each strain. The two S. cerevisiae strains with the best performance were selected for further fermentations on a Muscat Miel grape must, rich in bound monoterpenes. The levels of free linalool, hodiol I and geraniol increased significantly as compared to fermentation with a commercial wine yeast strain. These results show the suitability of this simple and economic medium to identify S. cerevisiae glucosidase producers with a potential impact on real winemaking conditions. On the other hand, great variability was found for the non-Saccharomyces strains, and this would demand further studies for each species. In conclusion, the use of EGA solid medium shows that the screening method is suitable for exploring the glucosidase activity of native strains of S. cerevisiae and shows good correlation with its real impact on free aroma compounds in the final wine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Adams RP (2001) Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy. Allured, Carol Stream

    Google Scholar 

  • Boido E, Lloret A, Medina K, Carrau F, Dellacassa E (2002) Effect of β-glucosidase activity of Oenococcus oeni on the glycosylated flavor precursors of Tannat wine during malolatic fermentation. J Agric Food Chem 50:2344–2349

    Article  CAS  Google Scholar 

  • Boido E, Lloret A, Medina K, Fariña L, Carrau F, Versini G, Dellacassa E (2003) Aroma composition of Vitis vinifera cv. Tannat: the typical red wine from Uruguay. J Agric Food Chem 51:5408–5413

    Article  CAS  Google Scholar 

  • Canal-Llauberes RS (1993) Enzymes in winemaking. In: Fleet GH (ed) Wine microbiology and biotechnology. Harwood Academic Publishers, Chur. Switzerland, pp 477–506

    Google Scholar 

  • Carrau F (1997) The emergence of a new Uruguayan wine industry. J Wine Res 8:179–185

    Article  Google Scholar 

  • Carrau F, Medina K, Boido E, Fariña L, Gaggero C, Dellacassa E, Versini G, Henschke PA (2005) de novo synthesis of monoterpenes by Saccharomyces cerevisiae wine yeasts. FEMS Microbiol Lett 243:107–115

    Article  CAS  Google Scholar 

  • Carrau F, Dellacassa E, Boido E (2008a) Terpenoids in grapes and wines: origin and micrometabolism during the vinification process. Nat Product Commun 3:577–592

    CAS  Google Scholar 

  • Carrau F, Medina K, Farina L, Boido E, Henschke PA, Dellacassa E (2008b) Production of fermentation aroma compounds by Saccharomyces cerevisiae wine yeast: effects of yeast assimilable nitrogen on two model strains. FEMS Yeast Res 8:1196–1208

    Article  CAS  Google Scholar 

  • Charoenchai C, Fleet GH, Henschke PA, Todd BEN (1997) Screening of non-Saccharomyces wine yeasts for the presence of extracellular hydrolytic enzymes. Aust J Grape Wine Res 3:2–8

    Article  CAS  Google Scholar 

  • Chassagne D, Guilloux-Benatier M, Alexandre H, Voilley A (2005) Sorption of wine volatile phenols by yeast lees. Food Chem 91:39–44

    Article  CAS  Google Scholar 

  • Ciani M, Comitini F, Mannazzu I, Domizio P (2010) Controlled mixed culture fermentation: a new perspective on the use of non-Saccharomyces yeasts in winemaking. FEMS Yeast Res 10:123–133

    Article  CAS  Google Scholar 

  • Cordero R, Ubeda J, Briones A, Potgieter N, Villena M, Pretorius I, Van Rensburg P (2003) Characterization of the β-glucosidase activity produced by enological strains of non-Saccharomyces yeast. J Food Sci 68:2564–2569

    Article  Google Scholar 

  • Darriet P, Boidron JN, Dubourdieu D (1988) L’hydrolyse des hétérosides terpéniques du Muscat á petit grains par les enzymes périplasmiques de Saccharomyces cerevisiae. Conn Vigne Vin 22:189–195

    CAS  Google Scholar 

  • Delcroix A, Gunata Z, Sapis JC, Salmon JM, Bayonove C (1994) Glycosidase activities of three enological yeast strains during winemaking: effect on the terpenol content of Muscat wine. Am J Enol Vitic 45:291–296

    CAS  Google Scholar 

  • Dubourdieu D, Darriet P, Ollivier C, Boidron JN, Ribéreau-Gayon P (1988) Role de la levure Saccharomyces cerevisiae dans l’hydrolyse enzymatique des hétérosides terpéniques du jus de raisin. Comt Ren Acad Sci Paris 306:489–493

    CAS  Google Scholar 

  • Eberhart B, Cross DF, Chase LR (1964) β-glucosidase system of Neurospora crassa. J Bacteriol 87:761–770

    CAS  Google Scholar 

  • Fernández M, Di Stefano R, Briones A (2003) Hydrolysis and transformation of terpene glycosides from muscat must by different yeast species. Food Microbiol 20:35–41

    Article  Google Scholar 

  • Fia G, Giovani G, Rosi I (2005) Study of β-glucosidase production by wine-related yeasts during alcoholic fermentation. A new rapid fluorimetric method to determine enzymatic activity. J Appl Microbiol 99:509–517

    Article  CAS  Google Scholar 

  • Fleet GH (2003) Yeast interactions and wine flavour. Int J Food Microbiol 86:11–22

    Article  CAS  Google Scholar 

  • Fowell RR (1965) The identification of wild yeast colonies on lysine agar. J Appl Bacteriol 28:373–383

    CAS  Google Scholar 

  • Goddard MR (2008) Quantifying the complexities of Saccharomyces cerevisiae’s ecosystem engineering via fermentation. Ecology 89:2077–2082

    Article  Google Scholar 

  • González-Pombo P, Pérez G, Carrau F, Carrascosa A, Guisán J, Batista-Viera F, Brena B (2008) One-step purification and characterization of an intracellular β-glucosidase of Metschnikowia pulcherrima. Biotechnol Lett 30:1469–1475

    Article  Google Scholar 

  • González-Techera A, Jubany S, Carrau F, Gaggero C (2001) Differentiation of industrial wine yeast strains using microsatellite markers. Lett Appl Microbiol 33:71–75

    Article  Google Scholar 

  • Guth H (1997) Quantitation and sensory studies of character impact odorants of different white wine varieties. J Agric Food Chem 45:3027–3032

    Article  CAS  Google Scholar 

  • Hernández LF, Espinosa JC, Fernández-González M, Briones A (2003) β-glucosidase activity in a Saccharomyces cerevisiae wine strain. Int J Food Microbiol 80:171–176

    Article  Google Scholar 

  • Jolly NP, Augustyn OPH, Pretorius IS (2003) The effect of non-Saccharomyces yeasts on fermentation and wine quality. S Afr J Enol Vitic 24:55–62

    CAS  Google Scholar 

  • Jolly NP, Augustyn OPH, Pretorius IS (2006) The role and use of non-Saccharomyces yeasts in wine production. S Afr J Enol Vitic 27:15–39

    CAS  Google Scholar 

  • Jubany S, Tomasco I, Ponce de León I, Medina K, Carrau F, Arrambide N, Naya H, Gaggero C (2008) Toward a global database for the molecular typing of Saccharomyces cerevisiae strains. FEMS Yeast Res 8:472–484

    Article  CAS  Google Scholar 

  • Kurtzman CP, Robnett CJ (1998) Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Ant van Leeuwen 73:331–371

    Article  CAS  Google Scholar 

  • Kwon K, Lee J, Kang HG, Hah YC (1994) Detection of β-glucosidase activity in polyacrylamide gels with esculin as substrate. Appl Environ Microbiol 60:4584–4586

    CAS  Google Scholar 

  • Mateo JJ, Di Stefano R (1997) Description of the β-glucosidase activity of wine yeasts. Food Microbiol 14:583–591

    Article  CAS  Google Scholar 

  • McLafferty FW, Stauffer DB (1991) The Wiley/NBS registry of mass spectral data, 5th edn. New York, Wiley

    Google Scholar 

  • McMahon HM, Zoecklein BW, Jasinski YW (1999) The effects of prefermentation maceration temperature and percent alcohol (v/v) at press on the concentration of Cabernet Sauvignon grape glycosides and glycoside fractions. Am J Enol Vitic 50:385–390

    CAS  Google Scholar 

  • Medina K, Boido E, Dellacassa E, Carrau F (2005) Yeast interactions with anthocyanins during red wine fermentation. Am J Enol Vitic 56:104–109

    CAS  Google Scholar 

  • Miskin A, Edberg SC (1978) Esculin hydrolysis reaction by Escherichia coli. J Clin Microbiol 7:251–254

    CAS  Google Scholar 

  • Pallmann CL, Brown JA, Olineka TL, Cocolin L, Mills DA, Bisson LF (2001) Use of WL Medium to profile native flora fermentations. Am J Enol Vitic 52:198–203

    CAS  Google Scholar 

  • Romano P, Suzzi G, Domizio P, Fatichenti F (1997) Secondary products formation as a tool for discriminating non-Saccharomyces wine strains. Ant van Leeuw 71:239–242

    Article  CAS  Google Scholar 

  • Rosi I, Vinella M, Domizio P (1994) Characterization of β-glucosidase activity in yeasts of oenological origin. J Appl Bacteriol 77:519–527

    CAS  Google Scholar 

  • Saqib AAN, Whitney PJ (2006) Esculin gel diffusion assay (EGDA): a simple and sensitive method for screening β-glucosidases. Enz Microb Technol 39:182–184

    Article  CAS  Google Scholar 

  • Soden A, Francis IL, Oakey H, Henschke PA (2000) Effects of co-fermentation with Candida stellata and Saccharomyces cerevisiae on the aroma and composition of Chardonnay wines. Aust J Grape Wine Res 6:21–30

    Article  CAS  Google Scholar 

  • Spagna G, Barbagallo R, Palmeri R, Restuccia C, Giudici P (2002) Properties of endogenous β-glucosidase of Saccharomyces cerevisiae strain isolated from Sicilian musts and wines. Enz Microb Technol 31:1030–1035

    Article  CAS  Google Scholar 

  • Strauss MLA, Jolly NP, Lambrechts MG, van Rensburg P (2001) Screening for the production of extracellular hydrolytic enzymes by non-Saccharomyces wine yeasts. J Appl Microbiol 91:182–190

    Article  CAS  Google Scholar 

  • Swiegers JH, Bartowsky PA, Henschke PA, Pretorius IS (2005) Yeast and bacterial modulation of wine aroma and flavour. Aust J Grape Wine Res 11:139–173

    Article  CAS  Google Scholar 

  • Vasserot Y, Christiaens P, Chimardin P, Arnaud A, Galzy P (1989) Purification and properties of a β-glucosidase of Hanseniaspora vineae Van der Walt and Tscheuschner with the view to its utilization in fruit aroma liberation. J Appl Bacteriol 66:271–279

    CAS  Google Scholar 

  • Versini G, Grando MS, Stefanini M, Dellacassa E, Carrau F (1999) Chemical, ampelometric and molecular methods applied for the identification of the uruguayan “Moscatel Miel” variety as the Italian “Moscato Giallo”. Quad Vitic Enol Univer

  • Williams PJ, Cynkar W, Francis L, Gray JD, Iland PG, Coombe BG (1995) Quantification of glycosides in grapes, juices and wines through a determination of glycosyl glucose. J Agric Food Chem 43:121–128

    Article  CAS  Google Scholar 

  • Yanai T, Sato S (2000) Purification and characterization of a novel α-L-arabinofuranosidase from Pichia capsulata X91. Bios Biotechnol Biochem 64:1181–1188

    Article  CAS  Google Scholar 

  • Zoecklein BW, Fugelsang K, Gump BH, Nury FS (1995) Wine analysis and production. Chapman & Hall, New York

    Google Scholar 

  • Zoecklein BW, Marcy JE, Williams JM, Jasinski Y (1997) Effect of native yeast and selected strains of Saccharomyces cerevisiae on Glycosyl Glucose, potential volatile terpenes, and selected aglycones of white Riesling (Vitis vinifera L.) wines. J Food Comp Anal 10:55–65

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was financed by Agencia Nacional de Investigacion e Innovacion (ANII), Instituto Nacional de Investigaciones Agricolas (INIA), Consejo Sectorial de Investigacion Cientifica (CSIC) and Programa de Desarrollo Tecnologico, (PDT 32-06) Uruguay. We would like to thank the critical comments and language revision made to this manuscript by Prof. Patrick Moyna.

Author information

Authors and Affiliations

  1. Enology Section, Food Science and Technology Department, Faculty of Chemistry, Universidad de la Republica, 11800, Montevideo, Uruguay

    Gabriel Pérez, Laura Fariña, Eduardo Boido & Francisco Carrau

  2. Molecular Biology Department, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), 11600, Montevideo, Uruguay

    Marianne Barquet & Carina Gaggero

  3. Chair of Farmacognosy and Natural Products, Organic Chemistry Department; Faculty of Chemistry, Universidad de la Republica, 11800, Montevideo, Uruguay

    Eduardo Dellacassa

Authors
  1. Gabriel Pérez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laura Fariña
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marianne Barquet
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Eduardo Boido
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carina Gaggero
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Eduardo Dellacassa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Francisco Carrau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Francisco Carrau.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pérez, G., Fariña, L., Barquet, M. et al. A quick screening method to identify β-glucosidase activity in native wine yeast strains: application of Esculin Glycerol Agar (EGA) medium. World J Microbiol Biotechnol 27, 47–55 (2011). https://doi.org/10.1007/s11274-010-0425-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-010-0425-4

Keywords

Search

Navigation