Elsevier

Food Chemistry

Volume 141, Issue 3, 1 December 2013, Pages 2513-2521
Food Chemistry

Increased flavour diversity of Chardonnay wines by spontaneous fermentation and co-fermentation with Hanseniaspora vineae

https://doi.org/10.1016/j.foodchem.2013.04.056Get rights and content

Highlights

  • Application of yeast Hanseniaspora vineae in real wine co-fermentation.

  • H. vineae and spontaneous fermentations showed higher flavour intensity and diversity.

  • Genetic strains identification showed increased diversity compared to conventional treatment.

  • Increased formation of esters and decrease formation of alcohols may explain fruit intensity.

Abstract

Discovery, characterisation and use of novel yeast strains for winemaking is increasingly regarded as a way for improving quality and to provide variation, including subtle characteristic differences in fine wines. The objective of this work was to evaluate the use of a native apiculate strain, selected from grapes, Hanseniaspora vineae (H. vineae) 02/5A. Fermentations were done in triplicate, working with 225 L oak barrels, using a Chardonnay grape must. Three yeast fermentation strategies were compared: conventional inoculation with a commercial Saccharomyces cerevisiae strain, ALG 804, sequential inoculation with H. vineae and then strain ALG 804 and spontaneous fermentation. Yeast strain identification was performed during fermentation, in which the apiculate strain was found to be active, until 9% of alcohol in volume, for the co-fermentation and the spontaneous fermentation was completed by three native S. cerevisiae strains. Basic winemaking parameters and some key chemical analysis, such as concentration of glycerol, biogenic amines, organic acids, and aroma compounds were analysed. Sensory analysis was done using a trained panel and further evaluated with professional winemakers. Sequential inoculation with H. vineae followed by S. cerevisiae resulted in relatively dry wines, with increased aroma and flavour diversity compared with wines resulting from inoculation with S. cerevisiae alone. Wines produced from sequential inoculations were considered, by a winemaker’s panel, to have an increased palate length and body. Characteristics of wines derived from sequential inoculation could be explained due to significant increases in glycerol and acetyl and ethyl ester flavour compounds and relative decreases in alcohols and fatty acids. Aroma sensory analysis of wine character and flavour, attributed to winemaking using H. vineae, indicated a significant increase in fruit intensity described as banana, pear, apple, citric fruits and guava. GC analysis of the relative accumulation of 23 compounds to significantly different concentrations for the three fermentation strategies is discussed in relation to aroma compound composition.

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

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