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Eur Food Res Technol (2014) 238:79–84DOI 10.1007/s00217-013-2062-2

ORIGINAL PAPER

Thermovinification of grapes from the Cabernet Sauvignon and Pinot Noir varieties using immobilized yeasts

Nathália de Andrade Neves · Lílian de Araújo Pantoja · Alexandre Soares dos Santos

Received: 26 April 2013 / Revised: 11 July 2013 / Accepted: 12 July 2013 / Published online: 28 August 2013 © Springer-Verlag Berlin Heidelberg 2013

Keywords Microvinification · Vitis vinifera · Diamantina · Wine

Introduction

Innovation in wine agribusiness may include multiple paths, such as diversification of the ‘terroir’ [1, 2], wine tourism [3], searching for new vinification processes [4], selecting or modifying fermenting microorganisms [5–7], use of forgotten grape varieties or those renewed by breed-ing, using oenological additives [8] or even the mischarac-terization of wine to obtain new products.

One of the indispensable elements of the wine produc-tion process is fermentation technology, which, combined with the intrinsic quality of the grapes, determines the extraction and stability of phenolic compounds such as fla-vonoids, tannins, anthocyanins and aromatic compounds present in grape skins and seeds and influences the compo-sition and quality of red wines. Among the technologies for wine production, thermovinification is an alternative to the traditional maceration processes for extracting dye com-pounds present in the skins of grapes. Advantages include the reduction in the microbial load of fruit, inhibition of undesirable enzymes such as polyphenol oxidase and lac-case, efficient extraction of anthocyanins and other poly-phenols and reduced time for winemaking [9–11]. Another intervention in the fermentation process, cell immobiliza-tion, has been applied to the production of alcoholic fer-mented beverages from fruit [12, 13], including still [14] and sparkling wines [15, 16]. This is a technique that, when applied to the production of fermented beverages, can increase productivity, modify the aroma profile, reduce the risk of contamination, allow for cell recycling and permit rapid clarification of the fermented mash [17].

Abstract This study examined the thermovinification technique using yeasts immobilized on calcium alginate for microvinification of the Cabernet Sauvignon and Pinot Noir grape varieties grown in Diamantina, Minas Gerais, Brazil. The wines obtained by thermovinification were characterized by their chemical and sensory aspects. The varietal wines from Cabernet Sauvignon and Pinot Noir had an alcohol content of 9.3 and 10.9 % (v/v), respec-tively. The total acidity, volatile acidity and the alcohol/reduced dry extract ratio were consistent with the Brazil-ian standards established for identity and quality of wines. There was a significant reduction in the levels of anthocya-nins, flavonoids and total phenolic compounds after aging in the bottle. The sensory evaluation presented an accept-ability exceeding 60 % for both wines.

N. de Andrade Neves Laboratory of Microvinification and Quality of Fermented Beverage-LabVin, Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM), Rodovia MGT 367, Km 583, No 5000-Alto da Jacuba, Diamantina, MG CEP 39100-000, Brazil

L. de Araújo Pantoja (*) Institute of Science and Technology, Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM), Rodovia MGT 367, Km 583, No 5000-Alto da Jacuba, Diamantina, MG CEP 39100-000, Brazile-mail: lilianpantoja@gmail.com

A. S. dos Santos Department of Basic Science, Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM), Rodovia MGT 367, Km 583, No 5000, Alto da Jacuba, Diamantina, MG CEP 39100-000, Brazile-mail: alexandre.soares@ufvjm.edu.br

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Considering the diversification of the ‘terroir’, Con-ceição and Tonieto [18] evaluated the climatic characteris-tics of Diamantina, Brazil, from the perspective of produc-ing grapes for the production of fine wines. According to these authors and the multicriteria climatic classification system [19], Diamantina presents the same climate classifi-cation as the viticulture region of the Serra Gaúcha, Brazil, during the spring–summer period and may also be included in the same climate group as the French wine regions of Bordeaux and Cognac during the fall–winter period. More-over, Diamantina is located in a region where the average altitude is 1,200 m.

The present study utilized thermovinification and the immobilization of yeast on calcium alginate for microvini-fication of the Cabernet Sauvignon and Pinot Noir varieties of the 2009 grape crop grown in Diamantina, Minas Gerais, Brazil, using a trellis system and also examined the phys-icochemical characteristics and acceptability of the wines produced.

Materials and methods

Cabernet Sauvignon and Pinot Noir grapes

The Cabernet Sauvignon and Pinot Noir grape varieties used in this work were planted in an experimental field in 2005 and grown using an trellis system on 1103 Paulsen rootstock. They were grown in an area at the elevation of 1,300 m and coordinates of 18°15′08″ south and 43°38′31″ west, in the city of Diamantina, state of Minas Gerais, Bra-zil. Grapes were harvested during the summer of 2009 in an advanced stage of maturation, after which they were packed in polypropylene boxes and then transported from the field to the laboratory where they were processed.

Thermovinification

About 10 kg of grapes of each variety was pressed in 6-mm-mesh nylon, the skins and seeds were separated and the juice was obtained. The cake was immersed in 10 % juice and underwent thermal extraction of the dye at 95 °C for 10 min. After cooling, this extract was added to the juice, making up the must.

For microvinification, a commercial dehydrated yeast was used (Saccharomyces cerevisiae—Fleischmann) immobilized with calcium alginate as reported by Pantoja et al. [12] in the proportion of 1,000 beads per liter of must. The fermentation process was carried out without agitation, at room temperature (25 ± 5 °C) and monitored at 12-h intervals.

At the end of the fermentation process, the fermented musts were transferred to 2-L PET bottles, supplemented

with 50 mg L−1 of potassium metabisulfite and maintained under refrigeration (4 ± 1 °C) for 30 days, followed by storage in amber glass bottles with a nominal capacity of 750 mL, closed with cork stoppers and stored in a dry, dark place for 6 months at an average temperature of 22 °C. After the period of aging in the bottle, chemical characteri-zation and acceptability testing were performed.

Chemical analysis

The total titratable acidity and the concentrations of solu-ble solids [20], anthocyanins (determined by the pH dif-ferential colorimetric method [21]), total flavonoids (deter-mined by spectrophotometry with aluminum chloride using quercetin as standard [22]) and total phenolic compounds (determined with Folin–Ciocalteau reagent with gallic acid as the standard [23]) were measured in the must from each variety of grape.

The fermentation processes were monitored at 12-h intervals for ethanol concentration (measured by the potas-sium dichromate spectrophotometric method using distilled samples [24]), total soluble solids, titratable acidity and pH.

The wine were analyzed for pH, titratable acidity, vola-tile acidity, dry extract, reduced dry extract, ash, total sul-fates, total alcohol–reduced dry extract ratio [20], reducing sugar and total reducing sugar [25], glycerol (determined using an enzymatic-colorimetric method with GPO Tri-glycerides Kit—Trinder Bio Liquid®), total sulfur dioxide by the Ripper method, total phenolic compounds, anthocy-anins, flavonoids and color intensity and hue [10].

Acceptability test

The acceptability test was conducted according to the method described by Chaves and Sproesser [26]. Tests were performed using questionnaires completed by 32 untrained adults, of which all were consumers of wine. Assessment of the degree of acceptability was conducted using a five-point hedonic scale where the panelists expressed how much they liked or disliked the wine. The tests were performed by offering approximately 50 mL of the wine, which was served separately in transparent acrylic cups at a tempera-ture of 15 ± 1 °C. Consumers were instructed on how to fill out the forms, and it was requested that they taste the wines and indicate the best option that reflected their judgment.

Results and discussion

Because of the high rainfall at harvest time, 236 mm for the month of January 2009 [27], the grapes were har-vested at an advanced stage of maturation in an attempt to concentrate the sugars in the grapes. Total soluble solid

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concentrations in the fruits were 17.20 ± 0.05 °Brix and 18.20 ± 0.02 °Brix for the Cabernet Sauvignon and Pinot Noir varieties, respectively. Thermal extraction of the pig-ments from the skins and seeds by cooking at 95 °C, char-acterizing a thermovinification process, resulted in extracts with volumes equivalent to about 20 % of the total must volumes. The extract obtained from the cake of Pinot Noir grapes presented a soluble solids concentration of 24.0 °Brix, while that obtained from the Cabernet Sauvignon cake was 21.5 °Brix. The main physicochemical character-istics of the musts for the grape varieties utilized, obtained by mixing the juices with their respective extracts, are sum-marized in Table 1.

The high acidity of the must from the Cabernet Sau-vignon grapes encountered in this work stands out as the relevant characteristic compared to the acidity of musts from grapes of the same variety grown in other regions of the country. Brunetto et al. [28] found values ranging from 42 to 72 mEq L−1 for musts of Cabernet Sauvignon grown in Campanha Gaucha, Rio Grande do Sul, Bra-zil, under different conditions of nitrogenation. Rizzon and Miele [29] found an average value for total acidity of 120 ± 16.3 mEq L−1 for musts from Cabernet Sauvignon grapes grown in Bento Gonçalves, Rio Grande do Sul, Bra-zil, from 1987 to 1992.

Vinification of Pinot Noir variety lasted 61 h (Fig. 1), after which the alcohol content reached 10.1 % (v/v) and TSS was reduced to 6 °Brix. In the vinification process of the Pinot Noir variety, the acclimation of the cells indicated a sigmoidal profile shown by the curves of sugar consump-tion (TSS) and alcohol production (Fig. 1).

For vinification of the Cabernet Sauvignon variety, acclimated beads were used, i.e., in their second cycle of usage. This acclimation resulted in progress curves for

sugar consumption and ethanol production with linear characteristics (Fig. 2). This response allows for the pos-sibility of reuse of biospheres to increase efficiency of the fermentation process. Fermentation of Cabernet Sauvignon ended with 5.2 °Brix, 11.5 % (v/v) ethanol concentration and reduction in total acidity from 171 to 134 mEq L−1.

The high microbial load used, 3.4 × 109 cells per liter of must, resulted in shorter fermentation times (ca. 65 h) than in traditional processes. The use of immobilized yeasts in fermentation processes has technical and economic advan-tages in comparison the use of free cells, particularly with regard to lower activation energy and higher reaction rate and characteristics of the immobilized cells, as well as the increased clarification speed of the beverages obtained.

Upon completion of the fermentation processes and clarification of the wine, which totaled 55 days, physico-chemical analyses were performed (Tables 2, 3). The wines were obtained with less than 5 g L−1 of reducing sugar and alcohol concentrations exceeding 9.3 % (v/v), which clas-sifies them as dry table wines. Values of total anthocyanins and color tone determined for the Cabernet Sauvignon and Pinot Noir wines presented a relationship consistent with

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anol

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Fig. 1 Vinification progress curve of the Pinot Noir variety

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Fig. 2 Vinification progress curve of the Cabernet Sauvignon variety

Table 1 Physicochemical characteristics of the musts obtained from the varieties of Vitis vinifera evaluated

Parameters Cabernet Sauvignon Pinot Noir

Total soluble solids (°Brix)

18.00 ± 0.05 18.50 ± 0.02

pH 3.49 3.49

Total titratable acidity (mEq L−1)

164.90 ± 0.68 129.98 ± 0.55

Total titratable acidity (Tartatic acid—g %)

1.24 0.97

TSS/tartatic acid 14.52 19.07

Potential alcohol (°Brix. 0.55) (% v/v)

9.90 10.17

Anthocyanins (mg L−1) 277.53 ± 11.57 95.51 ± 1.42

Flavonoids (mg L−1) 453.33 ± 4.71 374.57 ± 11.98

Phenolic compounds (mg L−1)

1044.00 ± 45.25 876.47 ± 38.82

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the pigmentation characteristics of each variety. The total acidity, volatile acidity, total sulfates, total sulfur dioxide and the ratio of ethanol content to the weight of reduced dry extract of the varietal wines made from Cabernet Sau-vignon and Pinot Noir met the current Brazilian regula-tions. There was also a significant reduction in total acidity among wines obtained after the clarification and cold sta-bilization periods (±4 °C), from 164.90 to 76.75 mEq L−1 for the Cabernet Sauvignon wine and from 129.98 to 96.19 mEq L−1 for the Pinot Noir wine.

The ash contents encountered were 2.05 g L−1 for Cab-ernet Sauvignon and 2.22 g L−1 for Pinot Noir, within the established values for ash according to Brazilian legislation which established a minimum of 1.5 g L−1. According to Rizzon and Miele [30], ash represents the mineral elements

present in wines. They interfere with the organoleptic char-acteristics, clarity, stability and pH of the beverages.

The glycerol concentrations of 5.72 and 6.58 g L−1 pre-sent in wines of the Cabernet Sauvignon and Pinot Noir varieties, respectively, are within the expected limits of 5–20 g L−1 as reported by Ribéreau-Gayon et al. [10] for red wines. Glycerol, a secondary product of alcoholic fer-mentation, is the second most important alcohol in wines. Sensorially, this is responsible for the sweet taste and con-tributes to the viscosity and texture of the wine [31].

After the aging period of 6 months in the bottle, there was a marked reduction in total anthocyanins, total flavo-noids and phenolic compounds in both wines (Table 3). On the other hand, there was an increased color intensity and tone in both wines. Monagas et al. [32] also reported

Table 2 Physicochemical parameters of the wines obtained by thermovinification of the Cabernet Sauvignon and Pinot Noir grapes after the clarification period

Parameters Cabernet Sauvignon Pinot Noir Legislation

Max. Min.

pH 3.48 ± 0.02 3.66 ± 0.02

Total titratable acidity (mEq L−1) 76.75 ± 0.74 96.19 ± 1.86 130 55

Total titratable acidity (Tartatic acid—g %) 5.8 7.2

Volatile acidity (mEq L−1) 0.23 ± 0.01 0.15 ± 0.00 20 –

Fixed acidity (mEq L−1) 76.16 ± 0.50 96.90 ± 1.57

Reduced dry extract (g L−1) 21.37 ± 1.10 18.92 ± 0.20

Ash (g L−1) 2.45 ± 0.13 2.24 ± 0.040 1.5

Total SO2 total (mg L−1) 15 ± 0.0 7.0 ± 0.0 350

Total sulfates (g L−1) <0.7 <0.7 1.0

Glycerol (g L−1) 3.9 ± 0. 8 5.2 ± 0.3

Ethanol % (v/v) 9.32 ± 0.16 10.89 ± 0.08 13 10

Reducing sugars (g L−1) 0.34 ± 0.00 0.44 ± 0.14

Total reducing sugars (g L−1) 0.74 ± 0.00 0.87 ± 0.18

Ethanol/reduced dry extract 3.44 4.54 5.2

Table 3 Physicochemical parameters related to the color of wines obtained by thermovinification of the Cabernet Sauvignon and Pinot Noir varieties before and after aging in the bottle

* Six months after bottling

Parameters Cabernet Sauvignon Pinot Noir

Before After* Before After*

Anthocyanins (mg L−1) 227.5 ± 2.1 65.5 ± 0.0 70.9 ± 0.5 23.4 ± 0.0

Flavonoids (mg L−1) 536.2 ± 23.9 156.8 ± 1.5 730.4 ± 24.2 204.2 ± 4.6

Phenolic compounds (mg L−1) 666.7 ± 20.9 256.7 ± 8.1 669.4 ± 32.7 271.4 ± 0.0

Reading at 420 nm 1.58 2.94 1.46 1.80

Reading at 520 nm 2.91 3.95 1.55 1.34

Reading at 620 nm 0.40 0.95 0.25 0.28

Color intensity (420 + 520 + 620 nm) 4.89 7.83 3.26 3.42

Tone (420/520 nm) 0.54 0.75 0.94 1.34

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a decrease in total anthocyanins from 340 to 216 mg L−1 and total flavonoids from 87.8 to 53.5 mg L−1 in Carbernet Sauvignon wines 6 months after bottling. The same authors observed increased absorption at the wavelength of 420 nm after 26 months of aging in the bottle. This is natural dur-ing the aging process of wines and is explained by the vari-ous reactions that result in degradation, modification and stabilization of color [33]. The thermovinification process, although it contributes to increased extraction of the dye compounds present in the skins of red grapes, especially anthocyanins, is not effective in extracting tannins, which are responsible for phenolic stability during aging of the beverage [34].

The acceptance of the wines was greater than 60 % (Table 4). However, the Pinot Noir variety received a higher score (63.2 %), with 45.95 % of the responses considered positive, which represent responses between liked and liked extremely on a scale from 1 to 5.

Conclusions

Use of the thermovinification technique associated with fermentation with immobilized yeasts significantly reduced the time required for wine production. However, the sta-bility of phenolic compounds was compromised, where there was a significant decrease in the concentrations of total anthocyanins and flavonoids. On the other hand, the acceptability test was not compromised by chemical altera-tions during aging in the bottle. The acceptability test with approval exceeding 60 % suggests that the technology might be adopted to a style of young and differentiated wines.

Conflict of interest None.

Compliance with Ethics Requirements This article does not con-tain any studies with human or animal subjects.

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Pinot Noir 63.2 29.73 45.95

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