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Article

Improved Saccharomyces cerevisiae Strain in Pure and Sequential Fermentation with Torulaspora delbrueckii for the Production of Verdicchio Wine with Reduced Sulfites

by
Alice Agarbati
,
Laura Canonico
*,
Francesca Comitini
and
Maurizio Ciani
Department of Life and Environmental Sciences, Polytechnic University of Marche, Via Brecce Bianche, 60131 Ancona, Italy
*
Author to whom correspondence should be addressed.
Appl. Sci. 2020, 10(19), 6722; https://doi.org/10.3390/app10196722
Submission received: 3 September 2020 / Revised: 16 September 2020 / Accepted: 24 September 2020 / Published: 25 September 2020
(This article belongs to the Special Issue Analysis in Beer and Wine)
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Abstract

:

Featured Application

The present research is finalized to propose a potential application of improved native Saccharomyces cerevisiae strain in pure and/or mixed fermentation with Torulaspora delbrueckii to obtain wine with a reduced sulfites content and a peculiar aromatic taste at winerycondition.

Abstract

The application of yeast strains that are low producers of sulfur compounds is actually required by winemakers for the production of organic wine. This purpose could be satisfied using a native Saccharomyces cerevisiae strain improved for oenological aptitudes. Moreover, to improve the aromatic complexity of wines, sequential fermentations carried out with S. cerevisiae/non-Saccharomyces yeast is widely used. For these reasons, in the present work an improved native S. cerevisiae low producer of sulfite and sulfide compounds was evaluated in pure and in sequential fermentation with a selected Torulaspora delbrueckii. Additionally, the influence of grape juices coming from three different vintages under winery conditions was evaluated. In pure fermentation, improved native S. cerevisiae strain exhibited a behavior related to vintage, highlighting that the composition of grape juice affects the fermentation process. In particular, an increase in ethyl octanoate (vintage 2017) and phenyl ethyl acetate (vintage 2018) was detected. Moreover, isoamyl acetate was highly consistent and could be a distinctive aroma of the strain. The sequential fermentation T. delbrueckii/S. cerevisiae determined an increase in aroma compounds such as phenyl ethyl acetate and ethyl hexanoate. In this way, it was possible to produce Verdicchio wine with reduced sulfites and characterized by a peculiar aromatic taste.

1. Introduction

The use of Saccharomyces cerevisiae as a starter culture significantly improves the control fermentation process, avoiding negative repercussions in winemaking. On the other hand, this biotechnological practice may determine a standardization of wine flavor [1]. Different approaches and alternative strategies to improve the analytical and aroma profile, enhancing the complexity of wines, were proposed. In this regard, several research studies evaluated the positive influence of non-Saccharomyces yeasts in mixed fermentation on the flavor complexity [2,3,4,5,6,7,8,9,10]. In addition, non-Saccharomyces yeasts exhibited other features such as bio-protective action, antimicrobial activity, and variations on the production of some compounds such as ethanol, glycerol, lactic acid in mixed fermentation that could be of interest in winemaking. In particular, Torulaspora delbrueckii showed the capacity to reduce the final ethanol content of wine and to act against spoilage yeasts [11,12,13].
The use of selected S. cerevisiae strains as starter cultures of fermentation could represent another possibility to confer a specific aromatic imprinting to the wine. This aspect could be further influenced by the metabolic interaction between S. cerevisiae/non-Saccharomyces strains in mixed starter cultures [14,15]. Indeed, S. cerevisiae strains can metabolically interact through the modification of fermentation products when they grow in mixed cultures. For this reason, in recent decades, researchers proposed the use of tailored strains, often isolated from grapes (native yeast strains) that possibly reflect a specific geographical area, as a valid alternative to the use of selected starter cultures, with the purpose of conferring uniqueness to the wines [16,17,18,19,20,21]. On the other hand, the native S. cerevisiae strains often do not show all the desired oenological aptitudes. About this, several studies described different approaches and strategies to improve S. cerevisiae strains obtaining yeasts with desired characteristics suitable for winemaking [21,22,23]. For these reasons, in the present study, we evaluated the use of the improved native S. cerevisiae for the low production of sulfite and sulfide compounds coming from Verdicchio wine area [21]. This S. cerevisiae strain was evaluated in pure and in sequential fermentation with a selected T. delbrueckii strain for the production of low sulfites wine under winery conditions. The fermentation performance and the aroma profile of the resulted wines were assessed.

2. Materials and Methods

2.1. Yeast Strains

The improved S. cerevisiae native strain, DiSVA 708, belonging to the Yeast Collection of the Department of Life and Environmental Sciences (DiSVA) of the Polytechnic University of Marche (Italy) [21] and the commercial S. cerevisiae starter strain, Lalvin ICV OKAY® (Lallemand Inc., Toulouse, France) were used. The commercial strain is characterized by the absence of H2S production and reduced production of SO2.
The non-Saccharomyces strain, T. delbrueckii DiSVA 130, was used in the sequential fermentations with the improved native S. cerevisiae DiSVA 708 over the two vintages (2017–2018). It was originally isolated from grapes and used in mixed fermentation to enhance the analytical and aromatic profile of the wine [24].
All of the yeast strains were maintained on yeast extract–peptone–dextrose (YPD) agar medium (Oxoid, Basingstoke, U.K.) at 4 °C for short-term storage and in YPD broth supplemented with 80% (w/v) glycerol at −80 °C for long-term storage.

2.2. Fermentation Trials

Fermentation trials were performed over three consecutive years: 2016–2018, using organic Verdicchio grape juice. For each vintage, the grapes were treated following the same winemaking process (soft hydraulic press pressing and cold clarification at 8 °C for 5 days). The analytical characters of the grape musts (initial sugars, pH, total acidity, malic acid, total SO2 and nitrogen content) are shown in Table 1. The diammonium phosphate and yeast derivative (Genesis Lift® Oenofrance, Bordeaux, France) used were adjusted to 250 mg N/L as yeast assimilable nitrogen. The fermentation trials were carried out in a steel vessel containing 40 L of grape juice in duplicate under static conditions at 18 °C.
All of the yeast strains were pre-cultured in modified YPD medium (0.5% yeast extract, 0.1% peptone and 2% glucose) for 72 h at 25 °C in an orbital shaker (150 rpm). The biomass was collected by centrifugation, washed three times with sterile distilled water and inoculated into the grape juice to obtain an initial concentration of approximately 1 × 106 cell mL−1. For vintages 2017 and 2018, the sequential fermentations were carried out by inoculation of T. delbrueckii DiSVA 130 followed by inoculation of S. cerevisiae DiSVA 708 after two days from the start of fermentation. The growth kinetics of the yeast strains were monitored during the fermentation at established intervals using WL nutrient Agar medium (Oxoid, Hampshire, U.K.) and Lysine Agar medium (Oxoid, Hampshire, U.K.) [25]. The sugar consumption during the fermentation process was measured by Baumé (°Bé) densimeter.

2.3. Analytical Procedures

Total acidity, volatile acidity, pH, ethanol content and total SO2 were determined according to the Official European Union Methods (EC Regulation No. 2870/00) [26]. Enzymatic kits (Megazyme International Ireland) were used to measure the amounts of glucose and fructose (K-FRUGL), glycerol (K-GCROL), and malic acid (K-DMAL) according to the manufacturer instructions. Acetate strips (CARLO ERBA Reagents S.r.l., Milan, Italy) were used to evaluate the H2S production during the fermentation process. The production of H2S was detected by the increase in color of acetate strips from white to black in function of H2S during the fermentation. The semi-quantitative evaluation was expressed using an arbitrary scoring scale from zero (no H2S production, white strip) to 5 (maximum level of H2S production, black strip).
A specific enzymatic kit (kit no. 112732; Roche Diagnostics, Germany) was used to determine the ammonium content. The free α-amino acids were evaluated following Dukes and Butzke protocol [27].
Ethyl acetate, acetaldehyde and alcohols (n-propanol, isobutanol, amyl and isoamyl alcohols) were quantified using a gas chromatograph system (GC-2014; Shimadzu, Kjoto, Japan) by direct injection. The final wines were prepared as described by Canonico et al. [24]. The solid-phase microextraction (HS-SPME) method was used to quantify the main volatile compounds as described by Canonico et al. [28]. The compounds were desorbed by inserting the fiber Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS) (Sigma-Aldrich, St. Louis, MO, USA) into a gas chromatograph (GC) injector.

2.4. Sensory Analysis

At the end of the fermentation, the wines obtained were transferred into completely filled 750 mL bottles, closed with the crown cap and maintained at 4 °C until sensory analysis. After storage for 3 months, wines were subjected to sensory analysis on the basis of smell and taste. A group of ten testers (80% expert and 20% non-expert), using a score scale of 1 to 10, expressed their opinion regarding smell and taste of each wine tested. The data obtained were used to compare the wines and provide information regarding the organoleptic quality and probable consumers’ acceptability of the wines obtained.

2.5. Statistical Analysis

Analysis of variance (ANOVA) was used to elaborate the analytical character data of wine. The means were analyzed using the statistical software package JMP® 11. Duncan tests were used to detect the significant differences. The experimental data were significant with associated p-values < 0.05.
Principal Component Analysis (PCA) carried out using JMP 11® statistical software (Statistical discovery from SAS, New York, NY, USA) was used to analyze the mean values of each volatile compound and the main by-products of fermentation. The mean data were normalized to eliminate the influence of hidden factors.

3. Results

3.1. Biomass Evolution and Sugar Consumption

Growth kinetics of the pure and sequential fermentations carried out during the vintages 2016–2018 are reported in Figure 1. The results showed that improved S. cerevisiae DiSVA 708 exhibited the same growth kinetics of OKAY® starter strain in vintage 2016 (Figure 1a), while in vintages 2017–2018, DiSVA 708 showed lower growth kinetics. In particular, DiSVA 708 achieved a similar biomass concentration (about 5 × 107 CFU/mL) exhibited by OKAY® on the 8th day and 5th day of fermentation and in vintage 2017 and 2018, respectively (Figure 1b,c). In sequential fermentations, T. delbrueckii on the 2nd day of fermentation (2017–2018) achieved the maximum cell concentration (about 107 cell/mL). In vintage 2017 (Figure 1b), this value was constant until the end of fermentation, while in vintage 2018 the cell concentration decreased until 106 cell/mL (Figure 1c). In both vintages (2017–2018), the DiSVA 708 strain, inoculated after 48 h of fermentation, reached on the 5th day of fermentation the maximum cell concentration and remained approximatively constant until the end of fermentation (about 5 × 107 cell/mL) (Figure 1b,c).
The sugar consumption (Figure 2) exhibited a similar trend independently by fermentation trials and vintages, completing the fermentation process on the 10th day. Only S. cerevisiae OKAY® showed a faster rate of sugar consumption in vintage 2016 (Figure 2).

3.2. Main Oenological Characters and Volatile Compounds of S. cerevisiae Pure Fermentations

The main analytical characters of the fermentation trials carried out during the three consecutive vintages are reported in Table 2. With regard to the ethanol content in the resulting wines carried out by the S. cerevisiae pure fermentations (vintages 2017–2018), the two strains exhibited a comparable alcohol content consuming all sugars. Only in vintage 2016, DiSVA 708 showed a lower ethanol content than that exhibited by OKAY® due to a slight amount of residual sugars. On the other hand, the differences in the final ethanol content among the three vintages were due to different initial sugar content in the grape juice. OKAY® fermentation trials showed a constantly lower total acidity than the DiSVA 708 pure fermentation trials through the vintages, while the production of volatile acidity showed more variation among the vintages although it was produced in low quantities. With regard to the glycerol content, S. cerevisiae fermentations did not show significant differences, with the exception of T. delbrueckii sequential fermentation (vintage 2017). A variable trend was exhibited for malic acid content.
The starter OKAY® exhibited the lowest SO2 production in all vintages, despite the variability found among the vintages. However, the SO2 content of the other fermentation trials was in any case low in relation to the initial SO2 content of grape juice. The score of H2S production was the same for both S. cerevisiae pure fermentation in vintages 2017–2018 while in 2016 it was significantly different.
The results of volatile compounds are reported in Table 3. DiSVA 708 led a greater general increase in esters content than did OKAY®, with a different trend among the vintages. In particular, DiSVA 708 significantly increased ethyl acetate (2016); ethyl hexanoate (2017); phenyl ethyl acetate, ethyl octanoate and isoamyl acetate (2018). On the other hand, S. cerevisiae OKAY® showed a significantly high production in ethyl butyrate in all vintages, a compound characterizing the volatile profile of this strain. With regard to higher alcohols, the variations found were more dependent on the harvest than on the inoculated strains. The production of n-propanol was an exception. Indeed, OKAY® showed a production of n-propanol consistently and was significantly higher than the other trials in all vintages even when at different concentrations. Notably, an increase in acetaldehyde content in pure fermentation in vintage 2016 and a reduction in vintage 2018 by DiSVA 708 were observed.

3.3. Main Oenological Characters and Volatile Compounds of T. delbrueckii/S. cerevisiae DiSVA 708 Sequential Fermentations

The results of the main oenological characters (Table 2) of T. delbrueckii/S. cerevisiae DiSVA 708 sequential fermentation highlighted a comparable trend with regard to ethanol, residual sugar, total and volatile acidity and malic acid in comparison to DiSVA 708 pure fermentation. Meanwhile, significant differences among the vintages were found to highlight the differences in grape juice composition between one vintage and the other. The use of T. delbrueckii in sequential fermentation allowed a greater reduction in SO2 content than in DiSVA 708 pure fermentation in both vintages (significant only vintage 2017) (Table 2). Moreover, T. delbrueckii sequential fermentations exhibited the same H2S production in vintages 2017–2018, but a higher score in comparison with S. cerevisiae pure cultures. T. delbrueckii sequential fermentations showed a significant reduction in glycerol content in comparison with the other trials while a variable trend of malic acid content was observed among the fermentation trials independently by strains and vintages.
The main by-products and the main volatile compounds are reported in Table 3. Sequential fermentation T. delbrueckii/S. cerevisiae DiSVA 708 showed a number of esters during vintage 2017 comparable with DiSVA 708 pure fermentation. In vintage 2018, the presence of T. delbrueckii led to an increase in phenyl ethyl acetate and ethyl hexanoate when compared with DiSVA 708 pure fermentation. Regarding the higher alcohol content, the use of T. delbrueckii led to wines with an increase in isoamyl alcohol and to a lesser extent of β-phenyl ethanol (vintage 2017). Monoterpenes compounds were generally improved by the presence and fermentation activity of T. delbrueckii, even if the geraniol in 2018 vintage was not detected. A significant reduction in acetaldehyde content was exhibited in vintage 2018.

3.4. Principal Component Analysis of the By-Products and Main Volatile Compounds

The PCA of the main volatile compounds and all by-products was used to assess the effect of vintage and the inoculated pure and sequential fermentations (Figure 3). The graphical representation indicated that the fermentations were grouped according to both vintage and inoculated S. cerevisiae. Indeed, it is possible to group for the vintages (bottom-right 2016; bottom-left 2017; upper 2018). Strain effect was also detected. S. cerevisiae OKAY® trials were grouped together in bottom quadrants. The sequential fermentations carried out with T. delbrueckii were grouped together in the upper-left quadrant and closed to DiSVA 708 pure fermentations of the same vintage that showed some variations. DiSVA 708 2016 trials were different from the other trials (2017–2018), probably caused by an incomplete fermentation (residual sugar). The results obtained by PCA analysis showed that the final composition of wine was affected at different levels by the vintage, inoculated S. cerevisiae and non-Saccharomyces used in sequential fermentation.

3.5. Sensory Analysis

The wines obtained by pure (OKAY® and S. cerevisiae DiSVA 708) and sequential fermentations (T. delbruekii DiSVA 130/S. cerevisiae DiSVA 708) underwent sensory analysis, and the results are reported in Table 4. All the wines obtained did not show significant differences regarding smell and taste. However, the testers expressed a positive judgement regarding each wine, characterized by specific aromatic notes and without defects.

4. Discussion

The wide use of a limited number of S. cerevisiae strains as starter cultures of fermentation could standardize the aroma and analytical profile of wine [1,29]. To overcome this problem, the use of selected native S. cerevisiae strains as starter cultures of fermentation could be a suitable strategy [21,22]. Another strategy to enhance the wine aroma complexity is the use of non-Saccharomyces yeasts in sequential fermentations. Indeed, several studies emphasize the use of non-Saccharomyces yeasts in winemaking for their attitudes to produce aromatic compounds (such as esters, terpenes, thiols) and modify some structural compounds as polysaccharides, volatile acidity and ethanol content [30,31,32,33,34,35,36].
In this work, we evaluated the fermentation performance of a native strain of S. cerevisiae (DiSVA 708) improved for the absence of H2S production and reduced production of SO2 [21] to be used in the production of organic wines with a low SO2 content. In pure fermentation, DiSVA 708 strain exhibited a behavior related to vintage, highlighting that the composition of grape juice can affect the fermentation performance of yeast strain. In particular, the increase in different aromatic compounds was detected among the vintages: in 2017 vintage ethyl octanoate, responsible for fruity flavors such as apple, pear and peach [37,38,39] and in 2018 vintage phenyl ethyl acetate, responsible for floral and honey aroma [38]. On the other hand, isoamyl acetate (banana flavors) was consistently highly produced by DiSVA 708 strain which can be considered a distinctive aroma of the strain. On the contrary, OKAY® showed a more homogenous wine aroma across all three vintages with ethyl butyrate and n-propanol as a distinctive aroma compound [40,41].
In sequential fermentation, the results showed an influence on analytical profiles of the Verdicchio wines by T. delbrueckii selected strain. In particular, the sequential fermentation carried out during the 2017 vintage showed a reduced content of volatile acidity than the wine obtained by the pure fermentation of the native S. cerevisiae DiSVA 708. The ability to reduce the volatile acidity by T. delbrueckii is a feature well documented [2,42,43]. Moreover, the use of T. delbrueckii in sequential fermentation led to a reduction in total SO2 content. Actually, there are no studies that directly correlate low sulfur dioxide production with the use of selected non-Saccharomyces yeasts. However, Bely and co-workers [42] highlighted the ability of T. delbrueckii to reduce SO2-binding compounds production. We found that T. delbrueckii in sequential fermentation reduced acetaldehyde content.
The sequential fermentation T. delbrueckii/S. cerevisiae determined a general increase in some aroma compounds affecting the aroma complexity of the wine, such as phenyl ethyl acetate, ethyl hexanoate and β-phenyl ethanol, even if not at significant levels. Several previous works highlighted the tendency of T. delbrueckii to form higher levels of acetate compounds, β-phenyl ethanol and a general improvement of the quality of wine aroma increasing the perception of varietal and fruity characters [15,44,45].
On the other hand, confirming a previous work [15], the aroma profile of resulting wines was affected not only by non-Saccharomyces/S. cerevisiae used in sequential fermentation, but also by the vintage.
Overall data emphasized the use of selected non-Saccharomyces yeasts, such as T. delbrueckii, in sequential fermentation with selected native S. cerevisiae strains for Verdicchio wine production. Indeed, on one side, the use of native S. cerevisiae reinforced the distinctive aromatic characteristics of a particular enological area, usually conferred by native yeast strains. On the other side, non-Saccharomyces yeast produced peculiar fermentation by-products that positively influenced the analytical profile of Verdicchio wine, characterized by a well-structured style. Furthermore, a suitable strategy to obtain wine with reduced sulfites content is the use of improved native S. cerevisiae DiSVA 708 (low sulfite producer) in pure and mixed fermentation with a selected T. delbrueckii strain.

Author Contributions

A.A., L.C., F.C. and M.C. contributed equally to the manuscript. All the authors participated in the design and discussion of the research. A.A. carried out the experimental part of the work. A.A. and L.C. carried out the analysis of the data. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding

Acknowledgments

The authors wish to thank the winery Terre Cortesi Moncaro s.r.c.l. for technical assistance.

Conflicts of Interest

The authors declare to not have financial and/or personal interests that could influence the study of the present manuscript.

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Applsci 10 06722 g001a 550Applsci 10 06722 g001b 550
Figure 1. Growth kinetics of the pure and sequential fermentations in three consecutive years: 2016 (a), 2017 (b) and 2018 (c). Pure cultures of S. cerevisiae Lalvin ICV OKAY® ( Applsci 10 06722 i001) and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i002) in each vintage. Sequential fermentations of T. delbrueckii DiSVA 130 ( Applsci 10 06722 i003) and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i004) in the 2017 and 2018 vintages.
Figure 1. Growth kinetics of the pure and sequential fermentations in three consecutive years: 2016 (a), 2017 (b) and 2018 (c). Pure cultures of S. cerevisiae Lalvin ICV OKAY® ( Applsci 10 06722 i001) and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i002) in each vintage. Sequential fermentations of T. delbrueckii DiSVA 130 ( Applsci 10 06722 i003) and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i004) in the 2017 and 2018 vintages.
Applsci 10 06722 g001aApplsci 10 06722 g001b
Applsci 10 06722 g002 550
Figure 2. Kinetics of sugar consumption of the pure and sequential fermentations of all vintages analyzed: 2016 (a), 2017 (b), 2018 (c). Pure cultures of S. cerevisiae Lalvin ICV OKAY® ( Applsci 10 06722 i001) and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i005) in each vintage. Sequential fermentations of T. delbrueckii DiSVA 130 and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i006) in the 2017 and 2018 vintages.
Figure 2. Kinetics of sugar consumption of the pure and sequential fermentations of all vintages analyzed: 2016 (a), 2017 (b), 2018 (c). Pure cultures of S. cerevisiae Lalvin ICV OKAY® ( Applsci 10 06722 i001) and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i005) in each vintage. Sequential fermentations of T. delbrueckii DiSVA 130 and S. cerevisiae DiSVA 708 ( Applsci 10 06722 i006) in the 2017 and 2018 vintages.
Applsci 10 06722 g002
Applsci 10 06722 g003 550
Figure 3. Principal Component Analysis based on the data regarding the main by-products of fermentation and volatile compounds of the wines obtained by pure and sequential fermentations for all vintages: 2016, 2017, 2018. The abbreviations Td/DiSVA 708 indicate the sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708.
Figure 3. Principal Component Analysis based on the data regarding the main by-products of fermentation and volatile compounds of the wines obtained by pure and sequential fermentations for all vintages: 2016, 2017, 2018. The abbreviations Td/DiSVA 708 indicate the sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708.
Applsci 10 06722 g003
Table
Table 1. Main analytical parameters of the organic Verdicchio grape juice used across the vintages 2016–2018.
Table 1. Main analytical parameters of the organic Verdicchio grape juice used across the vintages 2016–2018.
Grape Juice ParametersVintage 2016Vintage 2017Vintage 2018
Initial sugars (g L−1)236.00212.00203.00
pH3.223.223.26
Total acidity (g L−1)4.424.584.83
Malic acid (g L−1)2.702.502.40
Total SO2 (mg L−1)35.0027.0026.00
Nitrogen content YAN * (mg N L−1) 89.0060.0062.00
* YAN = yeast assimilable nitrogen.
Table
Table 2. Main analytical parameters of the wines obtained across the vintages 2016–2018. Data are means ± standard deviations. Values displaying different superscript letters (a,b,c,d,e) within each column are significantly different according to Duncan tests (p < 0.05). The abbreviations Td/708 indicate the sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708.
Table 2. Main analytical parameters of the wines obtained across the vintages 2016–2018. Data are means ± standard deviations. Values displaying different superscript letters (a,b,c,d,e) within each column are significantly different according to Duncan tests (p < 0.05). The abbreviations Td/708 indicate the sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708.
Ethanol (% v/v)Residual Sugars (g L−1)Total Acidity (Tartaric Acid g L−1)Volatile Acidity (Acetic Acid g L−1)Glycerol (g L−1)Malic Acid (g L−1)Total SO2 (mg L−1)H2S (Score)
2016OKAY14.24 ± 0.07 a2.13 ± 0.76 c5.73 ± 0.03 d0.54 ± 0.01 a7.48 ± 0.27 a1.35 ± 0.09 b6.50 ± 0.71 d1 ± 0.0 c
DiSVA 70813.89 ± 0.17 b7.01 ± 2.31 a6.17 ± 0.03 b0.49 ± 0.01 b6.82 ± 0.42 a1.60 ± 0.12 a18.00 ± 0.72 c3 ± 0.0 a
2017OKAY13.2 ± 0.12 c1.80 ± 0.14 c5.79 ± 0.12 d0.33 ± 0.05 d7.23 ± 0.19 a1.35 ± 0.07 b14.00 ± 2.83 c1 ± 0.0 c
DiSVA 70813.31 ± 0.04 c2.1 ± 0.28 c6.15 ± 0.10 b0.42 ± 0.03 c7.20 ± 0.24 a1.1 ± 0.00 c39.00 ± 2.82 a1 ± 0.0 c
Td/70813.26 ± 0.07 c3.60 ± 0.53 b6.01 ± 0.01 c0.30 ± 0.03 d5.23 ± 0.18 b1.0 ± 0.00 c33.50 ± 0.71 b3 ± 0.0 a
2018OKAY12.57 ± 0.01 d1.45 ± 0.07 d6.11 ± 0.03 b,c0.14 ± 0.01 eNA *1.65 ± 0.07 a24.00 ± 0.00 c2 ± 0.0 b
DiSVA 70812.43 ± 0.01 d1.55 ± 0.07 d6.40 ± 0.09 a0.15 ± 0.01 eNA1.55 ± 0.07 a33.00 ± 1.41 b2 ± 0.0 b
Td/70812.50 ± 0.02 d1.77 ± 0.00 c,d6.39 ± 0.03 a0.15 ± 0.01 eNA1.65 ± 0.07 a30.50 ± 2.12 b3 ± 0.0 a
NA * = not available.
Table
Table 3. The main by-products of fermentation and volatile compounds of the wines obtained for vintages 2016–2018, in pure and in sequential fermentations with the non-Saccharomyces strain added 2 days prior to the S. cerevisiae DiSVA 708 strain. Data are means ± standard deviations. Values displaying different superscript letters (a,b,c,d,e,f) within each line are significantly different according to Duncan tests (p < 0.05). The abbreviation Td/708 indicates the sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708.
Table 3. The main by-products of fermentation and volatile compounds of the wines obtained for vintages 2016–2018, in pure and in sequential fermentations with the non-Saccharomyces strain added 2 days prior to the S. cerevisiae DiSVA 708 strain. Data are means ± standard deviations. Values displaying different superscript letters (a,b,c,d,e,f) within each line are significantly different according to Duncan tests (p < 0.05). The abbreviation Td/708 indicates the sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708.
Esters (mg L−1)201620172018
OKAYDiSVA 708OKAYDiSVA 708Td/708OKAYDiSVA 708Td/708
Ethyl butyrate3.05 ± 0.08 a0.74 ± 0.07 b,c1.29 ± 0.66 b0.96 ± 0.05 b,c0.82 ± 0.01 b,c1.44 ± 0.54 b,c0.59 ± 0.01 c0.62 ± 0.00 c
Ethyl acetate23.14 ± 0.15 b31.14 ± 0.15 a12.74 ± 0.23 c13.97 ± 0.40 c12.92 ± 0.40 c18.18 ± 9.07 b,c24.25 ± 1.30 a,b16.92 ± 2.43 b,c
Phenyl ethyl acetate0.25 ± 0.00 c0.28 ± 0.04 c0.21 ± 0.04 c0.24 ± 0.01 c0.33 ± 0.01 b,c0.22 ± 0.07 c0.59 ± 0.08 a,b0.68 ± 0.06 a
Ethyl hexanoate0.43 ± 0.07 c0.25 ± 0.07 c0.84 ± 0.07 b1.69 ± 0.13 a1.70 ± 0.02 a0.36 ± 0.10 c0.23 ± 0.07 c0.71 ± 0.21 b
Ethyl octanoate0.01 ± 0.00 b0.01 ± 0.00 b0.00 ± 0.00 b0.01 ± 0.00 b0.01 ± 0.00 b0.009 ± 0.012 b0.02 ± 0.00 a0 ± 0 b
Isoamyl acetate1.16 ± 0.18 c2.14 ± 0.31 b1.71 ± 0.35 c2.88 ± 0.36 b3.15 ± 0.09 a,b2.30 ± 0.41 b3.97 ± 1.18 a3.83 ± 1.49 a
Alcohols (mg L−1)
n-propanol109.08 ± 0.31 b45.75 ± 0.01 e89.30 ± 0.47 c33.25 ± 0.11 f31.27 ± 0.60 f125.18 ± 4.98 a28.31 ± 0.74 d45.22 ± 1.18 e
Isobutanol13.19 ± 0.15 d,e23.93 ± 0.22 b12.76 ± 0.40 d,e11.74 ± 0.43 e20.43 ± 0.06 c14.79 ± 0.73 d28.31 ± 0.01 a27.78 ± 3.18 a
Amyl alcohol12.68 ± 0.17 c,d18.10 ± 0.44 a11.21 ± 0.30 e9.88 ± 0.09 f12.17 ± 0.95 d,e13.89 ± 0.01 b13.82 ± 0.35 b,c11.68 ± 0.89 d,e
Isoamyl alcohol116.46 ± 0.51 b,c151.91 ± 0.81 a120.78 ± 0.67 b64.53 ± 0.64 e110.11 ± 5.91 c114.23 ± 0.99 c60.36 ± 0.08 e82.72 ± 4.83 d
β-Phenyl ethanol12.03 ± 0.74 b,c16.61 ± 0.23 b14.22 ± 2.24 b,c8.32 ± 0.60 c12.97 ± 0.25 b,c29.38 ± 0.26 a27.86 ± 0.43 a29.86 ± 0.56 a
Carbonyl Compounds (mg L−1)
Acetaldehyde6.01 ± 0.1 d53.42 ± 0.27 a8.99 ± 0.23 c,d12.05 ± 0.01 c,d15.77 ± 0.45 c46.62 ± 10.47 a29.55 ± 0.91 b16.51 ± 1.87 c
Monoterpenes (mg L−1)
Linalool0.04 ± 0.04 a,b0.00 ± 0.00 c0.06 ± 0.01 a,b0.03 ± 0.01 b,c0.04 ± 0.00 b,c0.046 ± 0.015 a,b0.02 ± 0.001 b,c0.08 ± 0.02 a
Geraniol0.01 ± 0.01 b0.00 ± 0.00 c0.00 ± 0.00 c0.01 ± 0.01 b0.09 ± 0.02 a0.00 ± 0.00 c0.00 ± 0.00 c0.00 ± 0.00 c
Table
Table 4. Sensory analysis of the wines obtained by pure and sequential fermentations in three vintages. Pure culture was carried out by S. cerevisiae OKAY and DiSVA 708. The abbreviations Td/708 indicate sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708, respectively. Data are means ± standard deviations. Values displaying different superscript letters (a,b,c) within each column are significantly different according to Duncan tests (p < 0.05).
Table 4. Sensory analysis of the wines obtained by pure and sequential fermentations in three vintages. Pure culture was carried out by S. cerevisiae OKAY and DiSVA 708. The abbreviations Td/708 indicate sequential fermentations carried out by T. delbrueckii/S. cerevisiae DiSVA 708, respectively. Data are means ± standard deviations. Values displaying different superscript letters (a,b,c) within each column are significantly different according to Duncan tests (p < 0.05).
VintagesTrialsSmell (Score)Taste (Score)
2016OKAY6.78 ± 0.11 a,b6.34 ± 0.08 b
DiSVA 7087.35 ± 0.12 a7.55 ± 0.40 a
2017OKAY5.94 ± 0.03 c5.89 ± 0.76 c
DiSVA 7086.00 ± 0.10 c7.17 ± 0.16 a
Td/7085.78 ± 1.73 c6.66 ± 0.77 a,b
2018OKAY7.13 ± 0.52 a7.39 ± 1.6 a
DiSVA 7086.29 ± 1.78 b6.61 ± 1.22 a,b
Td/7087.05 ± 0.21 a6.82 ± 0.63 a,b

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Agarbati, A.; Canonico, L.; Comitini, F.; Ciani, M. Improved Saccharomyces cerevisiae Strain in Pure and Sequential Fermentation with Torulaspora delbrueckii for the Production of Verdicchio Wine with Reduced Sulfites. Appl. Sci. 2020, 10, 6722. https://doi.org/10.3390/app10196722

AMA Style

Agarbati A, Canonico L, Comitini F, Ciani M. Improved Saccharomyces cerevisiae Strain in Pure and Sequential Fermentation with Torulaspora delbrueckii for the Production of Verdicchio Wine with Reduced Sulfites. Applied Sciences. 2020; 10(19):6722. https://doi.org/10.3390/app10196722

Chicago/Turabian Style

Agarbati, Alice, Laura Canonico, Francesca Comitini, and Maurizio Ciani. 2020. "Improved Saccharomyces cerevisiae Strain in Pure and Sequential Fermentation with Torulaspora delbrueckii for the Production of Verdicchio Wine with Reduced Sulfites" Applied Sciences 10, no. 19: 6722. https://doi.org/10.3390/app10196722

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