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Most commercialized red wines are produced through alcoholic fermentation performed by yeasts of the Saccharomyces genus, and a second fermentation performed by lactic bacteria of the Oenococus oeni species once the first is completely finished. However, the classical process can suffer complications, of which the risks can increase in grape juices with high contents of sugar and pH. Due to climate change, these situations are becoming more common in the winemaking industry. The main risks in those scenarios are alcoholic-fermentation stops or sluggish and undesirable bacteria development while alcoholic fermentation is not finished yet and wine still contains residual sugars. The study propose a novel alternative that offers a solution or reduces the risk of those scenarios while increasing acidity, which is another serious problem of warm viticulture regions. The alternative consists of the combined use of Lachancea thermotolerans to reduce the pH of musts that suffer from a lack of acidity, Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) to achieve malic acid stability during the first stages of alcoholic fermentation, and Saccharomyces bayanus to complete the alcoholic fermentation in difficult wines of high potential alcohol degree of over 15% (v/v). The new proposed biotechnology produced wines with higher final concentrations in lactic acid, glycerol, color intensity, ethyl lactate and 2-phenyl ethyl acetate in 2.39 g/L, 0.52 g/L, 21%, 48% and 37% respectively than the classical methodology where Saccharomyces genus performs alcoholic fermentation and later Oenococus oeni performs malolactic fermentation. Additionally, the new alternative produced wines with lower concentration in ethanol, pH, acetic acid, ethyl acetate, diacetyl and 1-propanol in 0.37% (v/v), 0.26, 0.08 g/L, 22%, 69% and 28% respectively than the classic method.

Most commercialized red wines are produced through alcoholic fermentation performed by yeasts of the Saccharomyces genus, and a second fermentation performed by lactic bacteria of the Oenococus oeni species once the first is completely finished. However, the classical process can suffer complications, of which the risks can increase in grape juices with high contents of sugar and pH. Due to climate change, these situations are becoming more common in the winemaking industry. The main risks in those scenarios are alcoholic-fermentation stops or sluggish and undesirable bacteria development while alcoholic fermentation is not finished yet and wine still contains residual sugars. The study propose a novel alternative that offers a solution or reduces the risk of those scenarios while increasing acidity, which is another serious problem of warm viticulture regions. The alternative consists of the combined use of Lachancea thermotolerans to reduce the pH of musts that suffer from a lack of acidity, Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) to achieve malic acid stability during the first stages of alcoholic fermentation, and Saccharomyces bayanus to complete the alcoholic fermentation in difficult wines of high potential alcohol degree of over 15% (v/v). The new proposed biotechnology produced wines with higher final concentrations in lactic acid, glycerol, color intensity, ethyl lactate and 2-phenyl ethyl acetate in 2.39 g/L, 0.52 g/L, 21%, 48% and 37% respectively than the classical methodology where Saccharomyces genus performs alcoholic fermentation and later Oenococus oeni performs malolactic fermentation. Additionally, the new alternative produced wines with lower concentration in ethanol, pH, acetic acid, ethyl acetate, diacetyl and 1-propanol in 0.37% (v/v), 0.26, 0.08 g/L, 22%, 69% and 28% respectively than the classic method.

This study researched the winemaking performance of new biotechnology involving the cooperation of Lachancea and Schizosaccharomyces genera in the production of wine. In all fermentations where Lachancea thermotolerans was involved, higher lactic acid concentrations appeared, while all fermentations where Schizosaccharomyces pombe was involved, lower levels in malic acid concentration took place. The sensorial properties of the final wines varied accordingly. Differences in mouthfeel properties and acidity occurred in the different fermentation trials. Fermentations with the highest concentration of hydrolyzed mannose showed the highest mouthfeel properties, but the lack of acidity reduced their overall impression. Wines made from a combination of L. thermotolerans and S. pombe showed the highest overall impression and were preferred by the tasters due to the balance between mouthfeel properties and acidity.

This study researched the winemaking performance of new biotechnology involving the cooperation of Lachancea and Schizosaccharomyces genera in the production of wine. In all fermentations where Lachancea thermotolerans was involved, higher lactic acid concentrations appeared, while all fermentations where Schizosaccharomyces pombe was involved, lower levels in malic acid concentration took place. The sensorial properties of the final wines varied accordingly. Differences in mouthfeel properties and acidity occurred in the different fermentation trials. Fermentations with the highest concentration of hydrolyzed mannose showed the highest mouthfeel properties, but the lack of acidity reduced their overall impression. Wines made from a combination of L. thermotolerans and S. pombe showed the highest overall impression and were preferred by the tasters due to the balance between mouthfeel properties and acidity.

This study researched the winemaking performance of new biotechnology involving the cooperation of Lachancea and Schizosaccharomyces genera in the production of wine. In all fermentations where Lachancea thermotolerans was involved, higher lactic acid concentrations appeared, while all fermentations where Schizosaccharomyces pombe was involved, lower levels in malic acid concentration took place. The sensorial properties of the final wines varied accordingly. Differences in mouthfeel properties and acidity occurred in the different fermentation trials. Fermentations with the highest concentration of hydrolyzed mannose showed the highest mouthfeel properties, but the lack of acidity reduced their overall impression. Wines made from a combination of L. thermotolerans and S. pombe showed the highest overall impression and were preferred by the tasters due to the balance between mouthfeel properties and acidity.

Most wine aroma compounds, including the varietal fraction, are produced or released during wine production and derived from microbial activity. Varietal aromas, typically defined as terpenes and thiols, have been described as derived from their non-volatile precursors, released during wine fermentation by different yeast hydrolytic enzymes. The perception of these minority aroma compounds depends on the chemical matrix of the wine, especially on the presence of majority aroma compounds, such as esters or higher alcohols. Strategies aiming to reduce the production of these masking flavors are on the spotlight of enology research as a way to better define varietal standard profiles for the global market. Using a natural white must from Verdejo variety (defined as a thiol grape variety), here we describe the analytical and sensorial impact of using, in sequential inoculations, a selected strain of Metschnikowia pulcherrima, in combination with two different Saccharomyces cerevisiae strains. An increase in the levels of the thiol 4-MSP (4-methyl-4-sulfanylpentan-2-one) over its sensory threshold, together with a decrease in higher alcohol production, was observed when M. pulcherrima was used. This has an important impact on these wines, making them fruitier and fresher, always preferred by the sensory panel.

Most wine aroma compounds, including the varietal fraction, are produced or released during wine production and derived from microbial activity. Varietal aromas, typically defined as terpenes and thiols, have been described as derived from their non-volatile precursors, released during wine fermentation by different yeast hydrolytic enzymes. The perception of these minority aroma compounds depends on the chemical matrix of the wine, especially on the presence of majority aroma compounds, such as esters or higher alcohols. Strategies aiming to reduce the production of these masking flavors are on the spotlight of enology research as a way to better define varietal standard profiles for the global market. Using a natural white must from Verdejo variety (defined as a thiol grape variety), here we describe the analytical and sensorial impact of using, in sequential inoculations, a selected strain of Metschnikowia pulcherrima, in combination with two different Saccharomyces cerevisiae strains. An increase in the levels of the thiol 4-MSP (4-methyl-4-sulfanylpentan-2-one) over its sensory threshold, together with a decrease in higher alcohol production, was observed when M. pulcherrima was used. This has an important impact on these wines, making them fruitier and fresher, always preferred by the sensory panel.

This work examines the physiology of a new commercial strain of Torulaspora delbrueckii in the production of red wine following different combined fermentation strategies. For a detailed comparison, several yeast metabolites and the strains implantation were measured over the entire fermentation period. In all fermentations in which T. delbrueckii was involved, the ethanol concentration was reduced; some malic acid was consumed; more pyruvic acid was released, and fewer amounts of higher alcohols were produced. The sensorial properties of final wines varied widely, emphasising the structure of wine in sequential fermentations with T. delbrueckii. These wines presented the maximum overall impression and were preferred by tasters. Semi-industrial assays were carried out confirming these differences at a higher scale. No important differences were observed in volatile aroma composition between fermentations. However, differences in mouthfeel properties were observed in semi-industrial fermentations, which were correlated with an increase in the mannoprotein content of red wines fermented sequentially with T. delbrueckii.

Most red wines commercialized in the market use the malolactic fermentationprocess in order to ensure stability from a microbiological point of view. In this secondfermentation, malic acid is converted into L-lactic acid under controlled setups. Howeverthis process is not free from possible collateral effects that on some occasions produceoff-flavors, wine quality loss and human health problems. In warm viticulture regions suchas the south of Spain, the risk of suffering a deviation during the malolactic fermentationprocess increases due to the high must pH. This contributes to produce wines with highvolatile acidity and biogenic amine values. This manuscript develops a new red winemakingmethodology that consists of combining the use of two non-Saccharomyces yeast strains asan alternative to the traditional malolactic fermentation. In this method, malic acid is totallyconsumed by Schizosaccharomyces pombe, thus achieving the microbiological stabilizationobjective, while Lachancea thermotolerans produces lactic acid in order not to reduce andeven increase the acidity of wines produced from low acidity musts. This technique reducesthe risks inherent to the malolactic fermentation process when performed in warm regions.The result is more fruity wines that contain less acetic acid and biogenic amines than thetraditional controls that have undergone the classical malolactic fermentation.

This work examines the physiology of a new commercial strain of Torulaspora delbrueckii in the production of red wine following different combined fermentation strategies. For a detailed comparison, several yeast metabolites and the strains implantation were measured over the entire fermentation period. In all fermentations in which T. delbrueckii was involved, the ethanol concentration was reduced; some malic acid was consumed; more pyruvic acid was released, and fewer amounts of higher alcohols were produced. The sensorial properties of final wines varied widely, emphasising the structure of wine in sequential fermentations with T. delbrueckii. These wines presented the maximum overall impression and were preferred by tasters. Semi-industrial assays were carried out confirming these differences at a higher scale. No important differences were observed in volatile aroma composition between fermentations. However, differences in mouthfeel properties were observed in semi-industrial fermentations, which were correlated with an increase in the mannoprotein content of red wines fermented sequentially with T. delbrueckii.

Most red wines commercialized in the market use the malolactic fermentationprocess in order to ensure stability from a microbiological point of view. In this secondfermentation, malic acid is converted into L-lactic acid under controlled setups. Howeverthis process is not free from possible collateral effects that on some occasions produceoff-flavors, wine quality loss and human health problems. In warm viticulture regions suchas the south of Spain, the risk of suffering a deviation during the malolactic fermentationprocess increases due to the high must pH. This contributes to produce wines with highvolatile acidity and biogenic amine values. This manuscript develops a new red winemakingmethodology that consists of combining the use of two non-Saccharomyces yeast strains asan alternative to the traditional malolactic fermentation. In this method, malic acid is totallyconsumed by Schizosaccharomyces pombe, thus achieving the microbiological stabilizationobjective, while Lachancea thermotolerans produces lactic acid in order not to reduce andeven increase the acidity of wines produced from low acidity musts. This technique reducesthe risks inherent to the malolactic fermentation process when performed in warm regions.The result is more fruity wines that contain less acetic acid and biogenic amines than thetraditional controls that have undergone the classical malolactic fermentation.

This study discusses the optimisation of a selectiv e and differential medium which would facilitate the isolation of Schizosaccharomyces (a genus with a low incidence compared to other microorganisms) to select individuals from this genus for industrial purposes, especially in light of the recent approval of the use of yeasts from this genus in the wine industry by the International Organisation of Vine and Wine, or to detect the presence of such yeasts, for those many authors who consider them food spoilers. To this end, we studied various selective differential agents based on the main thephysiological characteristics of this species, such as its high resistance to high concentrations of sugar, sulfur dioxide, sorbic acid, benzoic acid, acetic acid or malo ethanolic fermentation. This selective medium is based on the resistance of the genus to the antibiotic actidione and its high resistance to inhibitory agents such as benzoic acid compared to possible microorganisms which can give rise to false positive results. Malic acid was used as a differential fact or due to the ability of this genus to metabolise it to ethanol, which allows detecting of the degradation of this compound. Lastly, the medium was successfully used to isolate strains of Schizosaccharomyces pombe from honey.

This study discusses the optimisation of a selectiv e and differential medium which would facilitate the isolation of Schizosaccharomyces (a genus with a low incidence compared to other microorganisms) to select individuals from this genus for industrial purposes, especially in light of the recent approval of the use of yeasts from this genus in the wine industry by the International Organisation of Vine and Wine, or to detect the presence of such yeasts, for those many authors who consider them food spoilers. To this end, we studied various selective differential agents based on the main thephysiological characteristics of this species, such as its high resistance to high concentrations of sugar, sulfur dioxide, sorbic acid, benzoic acid, acetic acid or malo ethanolic fermentation. This selective medium is based on the resistance of the genus to the antibiotic actidione and its high resistance to inhibitory agents such as benzoic acid compared to possible microorganisms which can give rise to false positive results. Malic acid was used as a differential fact or due to the ability of this genus to metabolise it to ethanol, which allows detecting of the degradation of this compound. Lastly, the medium was successfully used to isolate strains of Schizosaccharomyces pombe from honey.

The purpose of this work was to compare and optimise different selective and differential media to aid in isolating spoilage yeasts belonging to the Brettanomyces/Dekkera genera. Growth media containing selective and differential factors were employed. These were inoculated with strains of yeast representing Spanish oenological microbiota. Lastly, some of these isolation media were successfully applied in 24 types of wine with a high ethylphenol content, all of which were from the Haro Oenological Station (La Rioja, Spain). p-coumaric acid was determined using High performance liquid chromatography-photodiode-array detection-electrospray ionization mass spectrometry (HPLC-DAD-ESI/MS); 4-ethylphenol by using Solid phase micro extraction-gas chromatography-mass spectrometry (SPME-GC-MS); and the rest of the analysis was carried out using official OIV methodology. Actidione is the most effective selective factor for isolating Brettanomyces/Dekkera yeast genera. Other secondary selective factors (selective carbon sources, sorbic acid and ethanol as a microbicide agent) may be used successfully to eliminate potential false positivities; however, they slow growth and delay the time to obtain results.

This work studies the physiology of Schizosaccharomyces pombe strain 938 in the production of white wine with high malic acid levels as the sole fermentative yeast, as well as in mixed and sequential fermentations with Saccharomyces cerevisiae Cru Blanc. The induction of controlled maloalcoholic fermentation through the use of Schizosaccharomyces spp. is now being viewed with much interest. The acetic, malic and pyruvic acid concentrations, relative density and pH of the musts were measured over the entire fermentation period. In all fermentations in which Schizo. pombe 938 was involved, nearly all the malic acid was consumed and moderate acetic concentrations produced. The urea content and alcohol level of these wines were notably lower than in those made with Sacch. cerevisiae Cru Blanc alone. The pyruvic acid concentration was significantly higher in Schizo. pombe fermentations. The sensorial properties of the different final wines varied widely.

The purpose of this work was to compare and optimise different selective and differential media to aid in isolating spoilage yeasts belonging to the Brettanomyces/Dekkera genera. Growth media containing selective and differential factors were employed. These were inoculated with strains of yeast representing Spanish oenological microbiota. Lastly, some of these isolation media were successfully applied in 24 types of wine with a high ethylphenol content, all of which were from the Haro Oenological Station (La Rioja, Spain). p-coumaric acid was determined using High performance liquid chromatography-photodiode-array detection-electrospray ionization mass spectrometry (HPLC-DAD-ESI/MS); 4-ethylphenol by using Solid phase micro extraction-gas chromatography-mass spectrometry (SPME-GC-MS); and the rest of the analysis was carried out using official OIV methodology. Actidione is the most effective selective factor for isolating Brettanomyces/Dekkera yeast genera. Other secondary selective factors (selective carbon sources, sorbic acid and ethanol as a microbicide agent) may be used successfully to eliminate potential false positivities; however, they slow growth and delay the time to obtain results.

This work studies the physiology of Schizosaccharomyces pombe strain 938 in the production of white wine with high malic acid levels as the sole fermentative yeast, as well as in mixed and sequential fermentations with Saccharomyces cerevisiae Cru Blanc. The induction of controlled maloalcoholic fermentation through the use of Schizosaccharomyces spp. is now being viewed with much interest. The acetic, malic and pyruvic acid concentrations, relative density and pH of the musts were measured over the entire fermentation period. In all fermentations in which Schizo. pombe 938 was involved, nearly all the malic acid was consumed and moderate acetic concentrations produced. The urea content and alcohol level of these wines were notably lower than in those made with Sacch. cerevisiae Cru Blanc alone. The pyruvic acid concentration was significantly higher in Schizo. pombe fermentations. The sensorial properties of the different final wines varied widely.