Increased flavour diversity of Chardonnay wines by spontaneous fermentation and co-fermentation with Hanseniaspora vineae
Introduction
The limited number of commercial yeast strains, used for winemaking throughout the world, is thought to contribute to production of wines with relatively uniform style compromising potential diversity and the wine character. The situation is worsened by technical difficulties for implementing natural or spontaneous fermentation methodology at the winery level. Even though Saccharomyces cerevisiae produces most of the ethanol in wine, non-Saccharomyces yeasts (NS) present, play a significant role in producing aroma compounds, such as esters, higher alcohols, acids and monoterpenes (Romano et al., 1997, Swiegers et al., 2005), contributing with diversity of “flavour phenotypes”. The “flavour phenotype” is an interesting concept for yeast selection considering that now more than 1300 volatile compounds can be determined in wine (Cordente, Curtin, Varela, & Pretorius, 2012).
Relatively large populations of active S. cerevisiae are typically used for inoculation of wine musts in wine fermentation. Many studies have shown, however, that indigenous strains in must are not fully suppressed and can contribute significantly during the early stages of juice fermentation (Ciani et al., 2010, Fleet, 2008). NS yeasts are found predominantly on grapes, and to a lesser extent on cellar equipment (Zott, Miot-Sertier, Claisse, Lonvaud-Funel, & Masneuf-Pomarede, 2008). In what is termed “spontaneous fermentation”, there is a sequence of dominance by various NS grape must yeasts, followed by relatively alcohol tolerant S. cerevisiae which can then complete the fermentation (Fleet, 2003). Indigenous yeasts have been reported to contribute either positively or negatively to the overall sensory characteristics of wine. This suggests extensive diversity among NS strains, only some of which may be useful or beneficial for wine production. In addition, there have been only a limited number of studies in different laboratories with consistent methodologies for controlling available nutrients, a fact that was proved to affect yeast mixed culture fermentations (Carrau, 2003, Fleet and Heard, 1993).
Fermentation, using NS strains, has often been associated with higher levels of residual sugars and unpredictable by-products as well as off-flavour. However, a number of researchers and winemakers have also found that spontaneous fermentations are associated with greater wine body, unusual or odd aromas and flavours, creamy texture and greater complexity (Carrau, 2006, Fleet, 2003, Ramey, 1995, Varela et al., 2009). Only a few publications have provided support for these results, and to our knowledge all studies were done at the laboratory scale (Anfang et al., 2009, Carrau, 2003, Ciani et al., 2006, Ciani and Comitini, 2011, Egli et al., 1998, Fleet, 2003, Henick-Kling et al., 1998, Herraiz et al., 1990, Jolly et al., 2003, Pérez et al., 2011).
The controversy that remains regarding the organoleptic quality of wines, produced using different native apiculate yeasts, may reflect the relatively low level of occurrence of useful strains in the environment. Two earlier studies possibly discouraged development of NS apiculate yeasts in winemaking (Velázquez et al., 1991, Ciani and Picciotti, 1995). These workers found that fermentation using their strains resulted in production of large amounts of ethyl acetate and acetic acid. The main NS yeasts present on and in grape must are apiculate and include Kloeckera apiculata/Hanseniaspora uvarum, K. apis/H. guillermondii, K. africana/H. vineae, K. corticis/H. osmophila and K. javanica/H. occidentalis (Jolly, Augustyn, & Pretorius, 2006). These yeasts generally have moderate fermentative capacity. There is concern about the use of mixed cultures with these apiculates and commercial Saccharomyces, in that their initial growth may be inhibitory for subsequent S. cerevisiae growth, resulting in sluggish or stuck fermentations. However, in some cases, inhibition could be reversed by improving the specific nutrient availability during fermentation (Medina, Boido, Dellacassa, & Carrau, 2012).
Our initial screening of apiculate yeasts isolated from Tannat grapes resulted in the identification of about 5% as having good wine fermentation abilities. Wines produced using these strains had good sensory characteristics, including intense fruity aromas that contributed to varietal character using a sequentially mixed culture with commercial Saccharomyces (Carrau, 2006).
More than 500 NS strains were isolated from the Uruguayan winemaking environment over a 5 year period. Over 20 of these strains, isolated from grapes and the initial fermentation stages, were shown to contribute positively to aroma and to red and white wine quality (Jubany et al., 2008, Pérez et al., 2011). From this collection, we selected a strain of Hanseniaspora vineae (T02/5AF) to study sequential fermentation with a conventional Saccharomyces wine strain. The aim of this study was to compare and evaluate standard wine fermentation with spontaneous fermentation and co-fermentation with H. vineae and commercial Saccharomyces. We used a Chardonnay grape must under routine wine production conditions, by barrel fermentation, in which the main variable was the yeast used for fermentation.
Section snippets
Yeast strains
The commercial wine yeast strain used was S. cerevisiae ALG 804 (DSM, Denmark). The apiculate NS strain used was isolated from Tannat wine fermentation, H. vineae T02/5AF, that was recently genetically identified from other strains within the species (Barquet et al., 2012).
H. vineae was prepared for inoculation for fermentation by first growing 1 × 105 cells/ml of the strain for 12 h in YEPD (1% yeast extract 2% peptone, 2% glucose, 2% agar, containing 0.1 M citrate–phosphate buffer, pH 4.5). Then
Results
The time course for fermentation, using the three yeast fermentation conditions, is shown in Fig. 1. Inoculation with the commercial strain resulted in a typical fermentation, however, the rate was decreased significantly when H. vineae was inoculated onto the must for 6 days prior to addition of the commercial strain (Fig. 1A).
The slower fermentation rate, resulting from H vineae treatments, result in a cooler fermentation.
Fig. 1B highlights the time when MLF was complete in each case. It shows
Discussion
Acceptance for use of native yeast strains and mixed cultures in spontaneous fermentation has required decades of study (Ciani and Comitini, 2011, Fleet, 2008, Henschke, 1997). It is known that native yeast can improve wine complexity and quality, however there are well established risks associated with not inoculating grape musts at the beginning of vinification (Boulton, Singleton, & Bisson, 1996). The unknown abilities of diverse native strains to contribute to wine quality have greatly
Conclusions
The application of a selected strain of H. vineae as an inoculum starter, in a co-fermentation, was successfully used for the production of quality white barrel fermented Chardonnay wines. Furthermore, the wines produced had a uniquely fruity character, intense flavours, as well as full body and a relatively long palate length. To our knowledge this is the first report at the real winemaking level of a wine co-produced with apiculate yeast. One advantage for using a non-Saccharomyces starter is
Acknowledgements
We wish to thank the following agencies for financial support: CSIC Group Project 656, Sector Productivo Project 602 of University of the Republic UdelaR, Uruguay, and Bodegas Carrau R&D Project: New non-Saccharomyces strains for the industry; and Hanseniaspora vineae FMV Project of Faculty of Chemistry (ANII financial support).
We also thank Laboratorio Santa Elena S.A, for the preparation of yeast strains, Freixenet Laboratory in Cataluña for biogenic amines and organic acids analysis and the
References (47)
- et al.
Fermentation behaviour and metabolic interactions of multistarter wine yeast fermentations
International Journal of Food Microbiology
(2006) Yeast interactions and wine flavour
International Journal of Food Microbiology
(2003)- et al.
Growth of non-Saccharomyces yeasts affects nutrient availability for Saccharomyces cerevisiae during wine fermentation
International Journal of Food Microbiology
(2012) - et al.
Acetate ester formation in wine by mixed cultures in laboratory fermentations
International Journal of Food Microbiology
(2003) - et al.
Monitoring a mixed starter of Hanseniaspora vineae–Saccharomyces cerevisiae in natural must: Impact on 2-phenylethyl acetate production
International Journal of Food Microbiology
(2011) - et al.
Dynamics and diversity of non-Saccharomyces yeasts during the early stages in winemaking
International Journal of Food Microbiology
(2008) - et al.
Co-fermentation with Pichia kluyveri increases varietal thiol concentrations in Sauvignon blanc
Australian Journal of Grape and Wine Research
(2009) - et al.
Tandem repeat-tRNA (TRtRNA) PCR method for the molecular typing of non-Saccharomyces subspecies
Applied Microbiology and Biotechnology
(2012) - et al.
An improved means of monitoring malolactic fermentation in wines by TLC- densitometry
Journal of Planar Chromatography – Modern TLC
(1999) - et al.
Aroma composition of Vitis vinifera cv. Tannat: The typical red wine from Uruguay
Journal of Agricultural and Food Chemistry
(2003)
Principles and Practices of Wine Making
Production of fermentation aroma compounds by Saccharomyces cerevisiae wine yeasts: Effects of yeast assimilable nitrogen on two model strains
FEMS Yeast Research
Formation of biogenic amines as criteria for the selection of wine yeasts
World Journal of Microbiology and Biotechnology
Non-Saccharomyces wine yeasts have a promising role in biotechnological approaches to winemaking
Annals of Microbiology
Controlled mixed culture fermentation: A new perspective on the use of non-Saccharomyces yeasts in winemaking
FEMS Yeast Research
Oenological properties of non-Saccharomyces yeasts associated with wine-making
World Journal of Microbiology and Biotechnology
The growth kinetics and fermentation behaviour of some non-Saccharomyces yeasts associated with wine-making
Biotechnology Letters
Flavour-active wine yeasts
Appl Microbiol Biotechnol
Dynamics of indigenous and inoculated yeast populations and their effect on the sensory character of Riesling and Chardonnay wines
Journal of Applied Microbiology
Wine yeasts for the future
FEMS Yeast Research
Yeasts-growth during fermentation
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