*3.3.1 Selected yeasts*

*Advances in Grape and Wine Biotechnology*

observed at doses of up to 29 kV cm<sup>−</sup><sup>1</sup>

includes the infrared component [90]. The UV-C light (254 nm, 8.4 kJ m<sup>−</sup><sup>2</sup>

*3.2.3 Ultrasound*

*3.2.4 Pulsed light*

*3.2.5 e-Beam irradiation*

Sauvignon grapes with doses of 50 a 122 Hz, 5 kV cm<sup>−</sup><sup>1</sup>

treated with PEF, as obtained by Puértolas et al. [86] when treating Cabernet

At the level of grape-musts treated with PEF, adverse effects have not been

[87].

and anthocyanins and to increase the antioxidant capacity [79, 89].

treated berries, which facilitates their subsequent handling.

nins and vitisins have exhibited higher stability [81].

tion of anthocyanins from grape marc [91].

induced oxidation by e-beam irradiation [82].

final content of anthocyanins and flavanols [94].

cially with short expositions to ozone (<72 h, 30 μL L<sup>−</sup><sup>1</sup>

The ultrasound (US) treatment of red grape-musts is an effective alternative to keep the level of anthocyanins up as high as 97% [88]. This fact clearly shows that the US preserves the chemical stability of these pigments. Combinations of US with heat and ethanol can also be exploited to increase the extraction of total phenols

Pulsed light (PL) is a low-cost technological alternative with higher possibilities of being scaled to an industrial level than HHP, PEF, or e-beam irradiation [81]. Its efficacy varies as a function of the applied light's features. Thus, better performance is achieved with PL than with UV-C, since the former, in addition to its intensity,

duces micro-cracks in the skin of red grapes [90], inducing a high anthocyanin migration, although it is performed with lesser intensity than with HHP [74] or e-beam irradiation [82] and without affecting the external appearance of the

However, in wines obtained from red grapes treated with PL (12% UV-C, 10% UV-B, and 8% UV-A), a slight reduction of anthocyanins at doses of 10 pulses at 600 J has been noted. This may be associated with the oxidative degradation of these compounds by radiation [82]. Interestingly, vinylphenolic pyranoanthocya-

Electron beam (e-beam) irradiation can enhance the extraction of anthocyanins by up to 70% at 10 kGy [82], without affecting the external appearance of treated berries. Lower doses (0.5–3.0 kGy) have also shown improvements during extrac-

One disadvantage of this technology is the lowering of anthocyanin contents in the produced wines, as consequence of the induced oxidation by radiation [82]. Nonetheless, the content of vinylphenolic pyranoanthocyanins and vitisins is not affected due to the robustness of double bond in heteroaromatic ring under the

Grapes exposed to ozone have shown greater contents of flavanols and resveratrol [92, 93]. However, the continuous exposure of berries to this gas (30 μL L<sup>−</sup><sup>1</sup>

24 h) may produce skin hardening, causing slower extractions without affecting the

On the other hand, the efficacy of phenolic extraction has been related with the grape variety. Wines fabricated with grapes containing high level of flavanols (as Nebbiolo) improved their color stability during winemaking procedure, espe-

y, and 3.67 kJ kg<sup>−</sup><sup>1</sup>

, 15 min, 27°C) continuously applied pro-

.

,

) [95]. Accordingly, the

**194**

*3.2.6 Ozone*

The melatonin content can be increased by using *Saccharomyces* and *non-Saccharomyces* strains with high production of this compound [97], as an additional source to the melatonin coming from grapes [28]. However, some compounds like the phytosterols may be reduced during the winemaking process, since some *Saccharomyces* strains might be able to use them as nutrients [65]. Besides, contents of anthocyanins [98] and resveratrol [99] can diminish, as a result of being adsorbed by the yeast cell walls during the fermentation process.

Another issue to be aware during the winemaking process is the use of yeast with lower expression of anthocyanin-β-glucosidase activity, which is responsible for hydrolysis of anthocyanins [100].

### *3.3.2 Pyranoanthocyanins synthesis*

The most important are vinylphenolic pyranoanthocyanins and vitisins. They present high chemical stability due to the presence of a heteroaromatic fourth ring in their structure, formed by the integration of vinylphenols, pyruvate, or acetaldehyde in the structure of the anthocyanin precursor [101], which provides resistance against oxidation and discoloration in the presence of SO2 and/or increase of wine pH [102]. Moreover, pyranoanthocyanins possess microbiological stability, for instance, against *Dekkera/Brettanomyces*, since this yeast is not able to hydrolyze these pigments [103].

Fermentations with yeasts with hydroxycinnamate decarboxylase (HCDC+) activity have been studied as a strategy to improve the synthesis of vinylphenolic pyranoanthocyanins, from the condensation of anthocyanins with vinylphenols [101]. The vinylphenols are molecules released from hydroxycinnamic acids in grapes by the HCDC+ activity, which later on can serve as substrate to the synthesis of 4-ethylphenol by *Dekkera/Brettanomyces* [103]. By reducing the content of hydroxycinnamic acids, it is possible to prevent the synthesis of 4-ethylphenol and, in turn, the content of vinylphenolic pyranoanthocyanins can be increased.

Other interesting pyranoanthocyanin groups are the vitisins A and B, which arise from the condensation of pyruvic acid and acetaldehyde, respectively, together with the malvidin during or after the fermentation process [102].

Also, it is possible to increase vitisin A levels with *Schizosaccharomyces pombe* [104], of vinylphenolic pyranoanthocyanins in mixed fermentations of *S. cerevisiae* with *Pichia guilliermondii* [105] or by using species with high production of acetaldehyde, such as *Saccharomycodes ludwigii* [106], to improve the synthesis of vitisin B and other molecules with positive impact on the wine.

On the other hand, it is possible to enlarge the production of acetaldehyde by *S. cerevisiae* in the presence of metabolic inhibitors [71, 107], due to their effect on the alcohol dehydrogenase, which might enhance the synthesis of vitisin B.

#### **3.4 Post-fermentation strategies**

#### *3.4.1 Traditional aging of red wine*

The aging has direct effects on wine composition, since chemical and/or enzymatic oxidation processes, degradation of phenols on the presence of SO2, and

condensation and polymerization reactions [108], among others, take place at this stage, contributing to modify the content of bioactive compounds.

In general, anthocyanin, resveratrol, and flavonol levels tend to diminish with aging process [1, 108, 109]. So that, more benefits to health are attributed to young red wines. Regarding the resveratrol, hydrolysis of the glycosidic form and cis/trans isomerization take place [108], affecting its availability and activity.

At the same time, the content of pyranoanthocyanins increases through anthocyanin condensation with other molecules [101, 102]. Besides, the anthocyanic polymerization or anthocyanin-tannin condensation can be potentially increased.

Likewise, it can augment the content of monomeric flavanols from the hydrolysis of oligomeric and polymeric forms [1]. In fact, monomeric tannins possess high antioxidant capacity to act against free radicals and chelate metals [4, 5, 8], inhibit oxidative stress in cardiac hypertrophy cases, and inhibit cardiomyocyte apoptosis [110] as well as provide antimicrobial activity against oral pathogens [58].

#### *3.4.2 Aging on lees (AOL)*

In the last years, this aging technique has gained relevance in the production of red wine [109]. It consists of the release of polysaccharides from cell walls of selected yeasts lees toward the wine during its stay in barrel [111]. These released polysaccharides can enhance, among other attributes, the protection of phenolic compounds against oxidation, due to the lees that have higher oxygen affinity [112].

Nonetheless, it has been noted that anthocyanin contents can be reduced during AOL [111], especially within the first months of aging. This is a consequence of the adsorbent capacity of lees, particularly, cinnamic anthocyanins [109]. Although the loss of anthocyanins can be reduced with lees of species like *S'codes ludwigii* or *S. pombe* [111].

### **4. Additional considerations and future perspectives**

The protective effect ascribed to bioactive compounds from wine is not only related to only one compound but also to a combined effect of several of these compounds and to their interactions with other compounds present in food. Also, the moderate ingestion of wine is certainly an important factor.

Most studies have been conducted at preclinical levels (*in vitro* and i*n vivo*), aiming to elucidate the action mechanisms. Nonetheless, issues, including the absorption and bioconversion, the number of compounds and their subsequent metabolites in blood circulation, their accumulation and distribution on tissues, the chemical shapes capable of acting on specific receptors in the human organism, and so forth, are still not fully understood.

Despite the existing evidence, there is no consensus regarding its acceptance as an alternative, which aids in the prevention of diseases. Hence, more studies at the clinical level, considering a larger number of volunteers of different ethnicities, lifestyles, and health conditions, are certainly required, with the special consideration that these bioactive compounds cannot be used to replace the medicaments, since they do not possess curative properties, rather they are components of a healthy diet that can help to prevent diseases.

Within the potential strategies, some viticulture practices might contribute to improve the synthesis of bioactive compounds during the vine cultivation. Later into the winery, a proper extraction from the grapes, as well as procedures to

**197**

O2

*Improvement of the Bioactive Profile in Wines and Its Incidence on Human Health…*

minimize the loss of such compounds during the fermentation and aging stages, can

These kinds of technologies have demonstrated their efficacy to improve the extraction of bioactive compounds in pre-fermentation stages although, until now, some disadvantages have been reported during their application. For instance, the HHP, PL, and e-beam irradiation can diminish the content of anthocyanins like

In addition, the high extraction of vitisin derivatives at 70°C by using of HHP, as previously reported by Corrales et al. [80], converts the temperature into a critical parameter that limits its applicability in the winery. This fact indicates the need for

It has been observed that the antioxidant potential of wine may decrease in aged wines as a result of the reduction of anthocyanins, resveratrol, and flavonols and

In general, the vitisins have shown lower potential to neutralize free radicals like

The pyranoanthocyanin synthesis by incorporation of pyruvic acid in positions 4 and 5 of A-ring in the structure of the anthocyanin precursor can decrease the potential to suppress free radicals, which might be related to the loss of -OH from carbon 5, that together with -OH from carbon 7, favors the antioxidant activity of anthocyanins [114]. These condensations can be achieved at the fermentation level, although these mostly happen during the aging of wine. Thus, in accordance with the traditional winemaking process, these would be necessary as a strategy to

As in anthocyanin precursor state, pyranoanthocyanins have shown antioxidant

and anti-inflammatory activities. For example, against pro-oxidant (H2O2) and pro-inflammatory (TNF-α) molecules, in addition to neutralizing the secretion of interleukin 8 (IL-8) in cell cultivation of adenocarcinoma from the human colon [17]. Vitisin A has been shown a protective effect against the secretion of monocyte chemoattractant protein-1 (MCP-1) induced by TNF-α factor in human endothelial cell cultures [115], in addition to show great stability in simulated (*in vitro*) gastrointestinal conditions [116], indicating its potential availability and effectiveness in

<sup>−</sup> when com-

<sup>−</sup> with respect to their anthocyanin precursors [7], while the pyruvic adduct of

Likewise, during PL applications [81], it is important to ensure a uniform exposition of the berry surface. The authors suggest the use of roller conveyor belts to change the position of the irradiated berry in order to improve the extraction. Finally, the scaling of these technologies at the industrial level is still a pending issue since most studies have been carried out in small volumes and in static systems at laboratory level. In order to implement such technologies in wineries, more studies concerning large volumes and continuous flow systems, like the one performed

Another important issue is the presence of products such as alcohol-free wines in the markets, which have also shown effectiveness due to the high content of bioactive compounds but with the advantage of avoiding the problems associated with

*DOI: http://dx.doi.org/10.5772/intechopen.85861*

excessive ethanol ingestion.

**4.1 Emerging technologies**

cyanidin in treated grapes [78, 81, 82].

more studies to optimize the extraction process.

by González-Arenzana et al. [113] with PEF, are needed.

**4.2 Pyranoanthocyanins and their effects on health**

the simultaneous synthesis of condensation products.

pared with other pyranoanthocyanins.

*in vivo* conditions and at clinical level.

the delphinidin has shown greater ability to neutralize OH<sup>−</sup> and O2

provide physicochemical and microbiological stability to the wine.

improve the bioactive profile of produced wines.

#### *Improvement of the Bioactive Profile in Wines and Its Incidence on Human Health… DOI: http://dx.doi.org/10.5772/intechopen.85861*

minimize the loss of such compounds during the fermentation and aging stages, can improve the bioactive profile of produced wines.

Another important issue is the presence of products such as alcohol-free wines in the markets, which have also shown effectiveness due to the high content of bioactive compounds but with the advantage of avoiding the problems associated with excessive ethanol ingestion.

#### **4.1 Emerging technologies**

*Advances in Grape and Wine Biotechnology*

increased.

*3.4.2 Aging on lees (AOL)*

oxygen affinity [112].

*S. pombe* [111].

condensation and polymerization reactions [108], among others, take place at this

At the same time, the content of pyranoanthocyanins increases through anthocyanin condensation with other molecules [101, 102]. Besides, the anthocyanic polymerization or anthocyanin-tannin condensation can be potentially

Likewise, it can augment the content of monomeric flavanols from the hydrolysis of oligomeric and polymeric forms [1]. In fact, monomeric tannins possess high antioxidant capacity to act against free radicals and chelate metals [4, 5, 8], inhibit oxidative stress in cardiac hypertrophy cases, and inhibit cardiomyocyte apoptosis

In the last years, this aging technique has gained relevance in the production of red wine [109]. It consists of the release of polysaccharides from cell walls of selected yeasts lees toward the wine during its stay in barrel [111]. These released polysaccharides can enhance, among other attributes, the protection of phenolic compounds against oxidation, due to the lees that have higher

Nonetheless, it has been noted that anthocyanin contents can be reduced during AOL [111], especially within the first months of aging. This is a consequence of the adsorbent capacity of lees, particularly, cinnamic anthocyanins [109]. Although the loss of anthocyanins can be reduced with lees of species like *S'codes ludwigii* or

The protective effect ascribed to bioactive compounds from wine is not only related to only one compound but also to a combined effect of several of these compounds and to their interactions with other compounds present in food. Also,

Most studies have been conducted at preclinical levels (*in vitro* and i*n vivo*), aiming to elucidate the action mechanisms. Nonetheless, issues, including the absorption and bioconversion, the number of compounds and their subsequent metabolites in blood circulation, their accumulation and distribution on tissues, the chemical shapes capable of acting on specific receptors in the human organism,

Despite the existing evidence, there is no consensus regarding its acceptance as an alternative, which aids in the prevention of diseases. Hence, more studies at the clinical level, considering a larger number of volunteers of different ethnicities, lifestyles, and health conditions, are certainly required, with the special consideration that these bioactive compounds cannot be used to replace the medicaments, since they do not possess curative properties, rather they are components of a healthy diet

Within the potential strategies, some viticulture practices might contribute to improve the synthesis of bioactive compounds during the vine cultivation. Later into the winery, a proper extraction from the grapes, as well as procedures to

In general, anthocyanin, resveratrol, and flavonol levels tend to diminish with aging process [1, 108, 109]. So that, more benefits to health are attributed to young red wines. Regarding the resveratrol, hydrolysis of the glycosidic form and cis/trans

stage, contributing to modify the content of bioactive compounds.

isomerization take place [108], affecting its availability and activity.

[110] as well as provide antimicrobial activity against oral pathogens [58].

**4. Additional considerations and future perspectives**

the moderate ingestion of wine is certainly an important factor.

and so forth, are still not fully understood.

that can help to prevent diseases.

**196**

These kinds of technologies have demonstrated their efficacy to improve the extraction of bioactive compounds in pre-fermentation stages although, until now, some disadvantages have been reported during their application. For instance, the HHP, PL, and e-beam irradiation can diminish the content of anthocyanins like cyanidin in treated grapes [78, 81, 82].

In addition, the high extraction of vitisin derivatives at 70°C by using of HHP, as previously reported by Corrales et al. [80], converts the temperature into a critical parameter that limits its applicability in the winery. This fact indicates the need for more studies to optimize the extraction process.

Likewise, during PL applications [81], it is important to ensure a uniform exposition of the berry surface. The authors suggest the use of roller conveyor belts to change the position of the irradiated berry in order to improve the extraction.

Finally, the scaling of these technologies at the industrial level is still a pending issue since most studies have been carried out in small volumes and in static systems at laboratory level. In order to implement such technologies in wineries, more studies concerning large volumes and continuous flow systems, like the one performed by González-Arenzana et al. [113] with PEF, are needed.

## **4.2 Pyranoanthocyanins and their effects on health**

It has been observed that the antioxidant potential of wine may decrease in aged wines as a result of the reduction of anthocyanins, resveratrol, and flavonols and the simultaneous synthesis of condensation products.

In general, the vitisins have shown lower potential to neutralize free radicals like O2 <sup>−</sup> with respect to their anthocyanin precursors [7], while the pyruvic adduct of the delphinidin has shown greater ability to neutralize OH<sup>−</sup> and O2 <sup>−</sup> when compared with other pyranoanthocyanins.

The pyranoanthocyanin synthesis by incorporation of pyruvic acid in positions 4 and 5 of A-ring in the structure of the anthocyanin precursor can decrease the potential to suppress free radicals, which might be related to the loss of -OH from carbon 5, that together with -OH from carbon 7, favors the antioxidant activity of anthocyanins [114]. These condensations can be achieved at the fermentation level, although these mostly happen during the aging of wine. Thus, in accordance with the traditional winemaking process, these would be necessary as a strategy to provide physicochemical and microbiological stability to the wine.

As in anthocyanin precursor state, pyranoanthocyanins have shown antioxidant and anti-inflammatory activities. For example, against pro-oxidant (H2O2) and pro-inflammatory (TNF-α) molecules, in addition to neutralizing the secretion of interleukin 8 (IL-8) in cell cultivation of adenocarcinoma from the human colon [17]. Vitisin A has been shown a protective effect against the secretion of monocyte chemoattractant protein-1 (MCP-1) induced by TNF-α factor in human endothelial cell cultures [115], in addition to show great stability in simulated (*in vitro*) gastrointestinal conditions [116], indicating its potential availability and effectiveness in *in vivo* conditions and at clinical level.
