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The yeast Lachancea thermotolerans has the ability to produce notable amounts of lactic acid and reduce alcoholic strength in fermentation, so it has a considerable potential for mitigating negative impacts of climate changes in winemaking. In this study, a treatment with L. thermotolerans and Saccharomyces cerevisiae in sequential inoculation was compared to a control S. cerevisiae monoculture fermentation of Malvazija istarska (aka Malvasia Istriana) white grape must. Standard physico-chemical parameters of the obtained wines were determined by the OIV methods. Targeted (GC/FID and GC/MS) and untargeted (GC×GC/TOF-MS) gas chromatographic techniques were combined for the analysis of volatile compounds. Phenolic compounds were analyzed by UPLC/QqQ-MS/MS, and proteins by RP-HPLC-DAD, while a sensory analysis of wines was performed by a panel of trained and certified tasters. L. thermotolerans co-fermentation treatment increased the concentration of lactic acid and decreased alcoholic strength. L. thermotolerans increased the concentrations of geraniol, β-ionone, isobutanol, isobutyric acid, ethyl isobutyrate, several major acetates, ethyl lactate, and diethyl succinate, followed by many minor compounds. This wine also contained more hydroxycinnamoyl tartrates, while control S. cerevisiae wine had higher levels of free hydroxycinnamates. The effects on PR proteins were minor. L. thermotolerans co-fermentation slightly enhanced the sensory perception of tropical fruit, herbaceous, tobacco, and buttery odor notes, as well as fullness of body. With the largest number of identified volatile compounds up to date and other results obtained, this study contributes to the better understanding of oenological and especially aromatic potential of L. thermotolerans in white wine production. [ABSTRACT FROM AUTHOR]

Non-conventional yeasts are increasingly being used in the production of fermented beverages owing to their ability to create unique and high-quality products. The yeast Lachancea thermotolerans is of great industrial significance, particularly in the production of l (+)-lactic acid, which is beneficial for acidifying wine, beer, and potentially sake. To explore its potential in sake brewing, three L. thermotolerans strains were isolated from natural environments and their physiological and fermentative characteristics were examined. The isolates surpassed the L. thermotolerans type strain (NBRC 1985) in lactic acid production under various culture conditions and exhibited comparable growth rates to that of Saccharomyces cerevisiae at 15–20 °C. Sake brewing tests using these isolates yielded approximately 3500 ppm of lactic acid, with a slightly lower production of aroma components compared to that produced by sake yeast, and an ethanol content of approximately 11–12 % was obtained. Reverse transcription-quantitative polymerase chain reaction revealed variable expression in putative lactate dehydrogenase genes depending on the culture conditions. Our findings suggest that L. thermotolerans strains can be used in sake brewing to produce unique sake. [ABSTRACT FROM AUTHOR]

To date, there are no specific guidelines for the use of bioacidifying yeasts in winemaking. In this work, we aimed to characterize an oenological strain of Lachancea thermotolerans (Lt), a non-Saccharomyces lactic acid-producing yeast, and to test different sequential inoculation conditions with Saccharomyces cerevisiae (Sc). The results of bench scale vinifications showed that both the strategy of inoculating Sc 12 h after Lt and the mixing of Lt and Sc during fermentation delivered an acceptable increase in lactic acid (2 g/L) and a decrease in pH (about 0.15 units). Therefore, both strategies were implemented in winery experiments. Our results at the cellar scale showed no increase in acidity, which was likely due to the presence of indigenous yeasts. Overall, our experience shows the difficulty of translating laboratory protocols into cellar experiments and calls for further research into new strategies for implementing acidifying yeasts. [ABSTRACT FROM AUTHOR]

Most commercially available red wines undergo alcoholic fermentation by Saccharomyces yeasts, followed by a second fermentation with the lactic acid bacteria Oenococcus oeni once the initial process is complete. However, this traditional approach can encounter complications in specific scenarios. These situations pose risks such as stalled alcoholic fermentation or the growth of undesirable bacteria while the process remains incomplete, leaving residual sugars in the wine. To address these challenges and the issue of low acidity prevalent in warmer viticultural regions, several novel alternatives are available. The alternatives involve the combined use of Lachancea thermotolerans to increase the acidity of the musts, lactic acid bacteria (Oenococcus oeni and Lactiplantibacillus plantarum) to ensure malic acid stability during early alcoholic fermentation stages, and Saccharomyces cerevisiae to properly complete alcoholic fermentation. The study showed variations in the final chemical parameters of wines based on the microorganisms used.

Chitosan exerts a significant influence on various chemical parameters affecting the quality of wine produced using multiple strains of Lachancea thermotolerans. The impact of chitosan on these parameters varies depending on the specific strain studied. We observed that, under the influence of chitosan, the fermentation kinetics accelerated for all examined strains. The formation of lactic acid increased by 41% to 97% across the studied L. thermotolerans strains, depending on the specific strain. This effect also influenced acidity-related parameters such as total acidity, which increased by 28% to 60%, and pH, which experienced a decrease of over 0.5 units. The consumption of malic acid increased by 9% to 20% depending on the specific strain of L. thermotolerans. Nitrogen consumption also rose, as evidenced by all L. thermotolerans strains exhibiting a residual value of Primary Amino Nitrogen (PAN) of below the detection limit, and ammonia consumption increased by 90% to 100%, depending on the strain studied. However, certain parameters such as acetic acid, succinic acid, and glycerol showed contradictory results depending on the strain under investigation. In terms of volatile composition, chitosan supplementation led to increased production of i-butanol by 32% to 65%, 3-methylbutanol by 33% to 63%, and lactic acid ethyl ester by 58% to 91% across all studied strains of L. thermotolerans. Other analyzed aroma compounds exhibited varying changes depending on the specific strain of L. thermotolerans. [ABSTRACT FROM AUTHOR]

Assyrtiko is a rare ancient grape variety of Greece, which is known to produce Protected Designation of Origin (PDO) Santorini white wines. Besides the famous character of the volcanic terroir, Assyrtiko of Santorini is also marked by a low pH value and sharp acidity. The aim of the present study was to apply a new inoculation procedure that modulates the fermentation process by maintaining the unique sensorial characteristics of Assyrtiko wines based on acidity. For this purpose, the Lachancea thermotolerans species, known for the formation of lactic acid, was tested in sequential fermentation with three different Saccharomyces cerevisiae strains. At the end of the fermentation process, implantation control for S. cerevisiae strains (interdelta sequence profile analysis) was performed, oenological parameters were determined according to the OIV protocols, and the volatile compounds produced were measured by gas chromatography–mass spectrometry (GC/MS). Finally, all produced wines were evaluated by quantitative descriptive analysis by two groups of experts; the Greek team of oenologists from Santorini Island specialized in Assyrtiko wines, and the French team of oenologists specialized in wine from Bordeaux. As expected, the inoculated strain was the one that dominated the fermentation process, but nine S. cerevisiae indigenous strains were also identified in the produced wines. Lachancea thermotolerans produced 1 g/L of lactic and also modulated the volatile profile of the wines independently of the S. cerevisiae strain used. The origin of the panelists played an important role in bringing up sensorial traits, such as acidity. Our results led to a new interesting application of L. thermotolerans for white wine production adapted to climate change claims. [ABSTRACT FROM AUTHOR]

Increasingly high interest in yeast–yeast interactions in mixed-culture fermentation is seen along with beer consumers' demands driving both market growth and requests for biotechnological solutions that can provide better sensory characteristics. In this study, Lachancea thermotolerans and Saccharomyces cerevisiae with a cell population ratio of 10:1 were inoculated for sour beer fermentation while the process conditions within the brewing industry remained unchanged. With L. thermotolerans producing lactic acid (1.5–1.8 g/L) and bringing down the pH to 3.3–3.4 whilst adding no foreign flavors herein, this study revealed a new natural, fruity sour beer with a soft, sour taste. In this study, the double-yeast mixed-culture fermentation produced more flavor substances than a single-culture process, and plenty of isobutyl acetate and isoamyl acetate enhanced the fruit aroma and balanced the sour beer with a refreshing taste. While playing a positive role in improving the beer's quality, the double-yeast mixed-culture fermentation developed in this study helps to offer an alternative mass production solution for producing sour beer with the processes better controlled and the fermentation time reduced. The stress responses of the L. thermotolerans during the fermentation were revealed by integrating RNA sequencing (RNA-Seq) and metabolite data. Given that the metabolic flux distribution of the S. cerevisiae during the fermentation differed from that of the non-Saccharomyces yeasts, transcriptional analysis of non-Saccharomyces yeast and S. cerevisiae could be suitable in helping to develop strategies to modulate the transcriptional responses of specific genes that are associated with the aroma compounds released by S. cerevisiae and non-Saccharomyces yeasts. In the case of some non-Saccharomyces yeast species/strains, the diversion of alcoholic fermentation and the formation of a great number of secondary compounds may, in part, account for the low ethanol yield. [ABSTRACT FROM AUTHOR]

The yeast Lachancea thermotolerans has the ability to produce notable amounts of lactic acid and reduce alcoholic strength in fermentation, so it has a considerable potential for mitigating negative impacts of climate changes in winemaking. In this study, a treatment with L. thermotolerans and Saccharomyces cerevisiae in sequential inoculation was compared to a control S. cerevisiae monoculture fermentation of Malvazija istarska (aka Malvasia Istriana) white grape must. Standard physico-chemical parameters of the obtained wines were determined by the OIV methods. Targeted (GC/FID and GC/MS) and untargeted (GC×GC/TOF-MS) gas chromatographic techniques were combined for the analysis of volatile compounds. Phenolic compounds were analyzed by UPLC/QqQ-MS/MS, and proteins by RP-HPLC-DAD, while a sensory analysis of wines was performed by a panel of trained and certified tasters. L. thermotolerans co-fermentation treatment increased the concentration of lactic acid and decreased alcoholic strength. L. thermotolerans increased the concentrations of geraniol, β-ionone, isobutanol, isobutyric acid, ethyl isobutyrate, several major acetates, ethyl lactate, and diethyl succinate, followed by many minor compounds. This wine also contained more hydroxycinnamoyl tartrates, while control S. cerevisiae wine had higher levels of free hydroxycinnamates. The effects on PR proteins were minor. L. thermotolerans co-fermentation slightly enhanced the sensory perception of tropical fruit, herbaceous, tobacco, and buttery odor notes, as well as fullness of body. With the largest number of identified volatile compounds up to date and other results obtained, this study contributes to the better understanding of oenological and especially aromatic potential of L. thermotolerans in white wine production.

Botrytis cinerea poses a recurring threat to viticulture, causing significant yield losses each year. The study explored the biocontrol capabilities of commercially used winemaking yeasts as a strategy to manage B. cinerea in grape berries. The winemaking yeast strains— Saccharomyces cerevisiae ES181, Saccharomyces pastorianus KBG6, S. cerevisiae BCS103, Lachancea thermotolerans Omega, and Torulaspora delbrueckii TD291-reduced B. cinerea growth and conidiation in vitro. Furthermore, they demonstrated a decreased disease severity and number of conidia in grape berries. Among these strains, S. cerevisiae BCS103 was the most effective, inducing the expression of the defense-related gene PR4 in berries. Its diffusible compounds and volatile organic compounds also reduced the expression of BcLTF2 , a positive regulator of B. cinerea conidiogenesis. The examined winemaking yeast strains, especially S. cerevisiae BCS103, demonstrated effective inhibition of B. cinerea in vitro and in grape berries, influencing key defense genes and reducing BcLTF2 expression, offering potential solutions for disease management in viticulture. The study underscores the promise of commercially available winemaking yeast strains as eco-friendly tools against B. cinerea in viticulture. Leveraging their safety and existing use in winemaking offers a potential avenue for sustainable disease management. [ABSTRACT FROM AUTHOR]

One of the most notable effects of climate change, especially in warm regions, is the decrease in acidity (i.e., increase in pH) of wines and a reduction in their aromatic profile. To address this issue, must from a white grape variety with low acidity were inoculated with two non-Saccharomyces yeasts (Lachancea thermotolerans and Torulaspora delbrueckii) to enhance the acidity of the resulting wines. Basic oenological variables and major volatile compounds and polyols of the wines were analyzed, and the results were compared with those obtained through a Saccharomyces cerevisiae strain. Through multiple regression analysis, we found relations between the production of lactic acid to compounds involved in yeast metabolism and redox balance, including glycerol, acetic acid, isobutanol, isoamyl alcohols, and 2-phenylethanol. By means of principal component analysis, we obtained three components that explain more than 89% of the observed variability. The first component differentiates wines produced by L. thermotolerans; the second differentiates wines obtained by S. cerevisiae from those obtained by T. delbrueckii; and the third component is related to the temperature of fermentation. Organoleptic wines produced with S. cerevisiae were the best valuated, but taste was a highlight of the wines produced with L. thermotolerans due to possessing the best acidity. [ABSTRACT FROM AUTHOR]

To date, there are no specific guidelines for the use of bioacidifying yeasts in winemaking. In this work, we aimed to characterize an oenological strain of Lachancea thermotolerans (Lt), a non-Saccharomyces lactic acid-producing yeast, and to test different sequential inoculation conditions with Saccharomyces cerevisiae (Sc). The results of bench scale vinifications showed that both the strategy of inoculating Sc 12 h after Lt and the mixing of Lt and Sc during fermentation delivered an acceptable increase in lactic acid (2 g/L) and a decrease in pH (about 0.15 units). Therefore, both strategies were implemented in winery experiments. Our results at the cellar scale showed no increase in acidity, which was likely due to the presence of indigenous yeasts. Overall, our experience shows the difficulty of translating laboratory protocols into cellar experiments and calls for further research into new strategies for implementing acidifying yeasts.

The study explores diverse strains of Lachancea thermotolerans in single-inoculum wine fermentation conditions using synthetic grape must. It aims to analyze the role of the species without external influences like other microorganisms or natural grape must variability. Commercial strains and selected vineyard isolates, untested together previously, are assessed. The research evaluates volatile and non-volatile chemical compounds in final wine, revealing significant strain-based variations. L. thermotolerans notably produces lactic acid and consumes malic acid, exhibiting moderate ethanol levels. The volatile profile displays strain-specific impacts, affecting higher alcohol and ester concentrations compared to S. cerevisiae. These effects vary based on the specific compounds. Using a uniform synthetic must enables direct strain comparisons, eliminating grape-related, environmental, or timing variables in the experiment, facilitating clearer insights into the behavior of L. thermotolerans in wine fermentation. The study compares for the first time all available commercial strains of L. thermotolerans.

In the current situation, wine areas are affected by several problems in a context of global warming: asymmetric maturities, pH increasing, high alcohol degree and flat wines with low freshness and poor aroma profile. The use of emerging biotechnologies allows to control or manage such problems. Emerging non-Saccharomyces as Lachancea thermotolerans are very useful for controlling pH by the formation of stable lactic acid from sugars with a slight concomitant alcohol reduction. Lower pH improves freshness increasing simultaneously microbiological stability. The use of Hanseniaspora spp. (specially H. vineae and H. opuntiae) or Metschnikowia pulcherrima promotes a better aroma complexity and improves wine sensory profile by the expression of a more complex metabolic pattern and the release of extracellular enzymes. Some of them are also compatible or synergic with the acidification by L. thermotolerans, and M. pulcherrima is an interesting biotool for reductive winemaking and bioprotection. The use of bioprotection is a powerful tool in this context, allowing oxidation control by oxygen depletion, the inhibition of some wild microorganisms, improving the implantation of some starters and limiting SO2. This can be complemented with the use of reductive yeast derivatives with high contents of reducing peptides and relevant compounds such as glutathione that also are interesting to reduce SO2. Finally, the use of emerging non-thermal technologies as Ultra High-Pressure Homogenization (UHPH) and Pulsed Light (PL) increases wine stability by microbial control and inactivation of oxidative enzymes, improving the implantation of emerging non-Saccharomyces and lowering SO2 additions. [ABSTRACT FROM AUTHOR]

Assyrtiko is a rare ancient grape variety of Greece, which is known to produce Protected Designation of Origin (PDO) Santorini white wines. Besides the famous character of the volcanic terroir, Assyrtiko of Santorini is also marked by a low pH value and sharp acidity. The aim of the present study was to apply a new inoculation procedure that modulates the fermentation process by maintaining the unique sensorial characteristics of Assyrtiko wines based on acidity. For this purpose, the Lachancea thermotolerans species, known for the formation of lactic acid, was tested in sequential fermentation with three different Saccharomyces cerevisiae strains. At the end of the fermentation process, implantation control for S. cerevisiae strains (interdelta sequence profile analysis) was performed, oenological parameters were determined according to the OIV protocols, and the volatile compounds produced were measured by gas chromatography–mass spectrometry (GC/MS). Finally, all produced wines were evaluated by quantitative descriptive analysis by two groups of experts; the Greek team of oenologists from Santorini Island specialized in Assyrtiko wines, and the French team of oenologists specialized in wine from Bordeaux. As expected, the inoculated strain was the one that dominated the fermentation process, but nine S. cerevisiae indigenous strains were also identified in the produced wines. Lachancea thermotolerans produced 1 g/L of lactic and also modulated the volatile profile of the wines independently of the S. cerevisiae strain used. The origin of the panelists played an important role in bringing up sensorial traits, such as acidity. Our results led to a new interesting application of L. thermotolerans for white wine production adapted to climate change claims.

In the current situation, wine areas are affected by several problems in a context of global warming: asymmetric maturities, pH increasing, high alcohol degree and flat wines with low freshness and poor aroma profile. The use of emerging biotechnologies allows to control or manage such problems. Emerging non-Saccharomyces as Lachancea thermotolerans are very useful for controlling pH by the formation of stable lactic acid from sugars with a slight concomitant alcohol reduction. Lower pH improves freshness increasing simultaneously microbiological stability. The use of Hanseniaspora spp. (specially H. vineae and H. opuntiae) or Metschnikowia pulcherrima promotes a better aroma complexity and improves wine sensory profile by the expression of a more complex metabolic pattern and the release of extracellular enzymes. Some of them are also compatible or synergic with the acidification by L. thermotolerans, and M. pulcherrima is an interesting biotool for reductive winemaking and bioprotection. The use of bioprotection is a powerful tool in this context, allowing oxidation control by oxygen depletion, the inhibition of some wild microorganisms, improving the implantation of some starters and limiting SO2. This can be complemented with the use of reductive yeast derivatives with high contents of reducing peptides and relevant compounds such as glutathione that also are interesting to reduce SO2. Finally, the use of emerging non-thermal technologies as Ultra High-Pressure Homogenization (UHPH) and Pulsed Light (PL) increases wine stability by microbial control and inactivation of oxidative enzymes, improving the implantation of emerging non-Saccharomyces and lowering SO2 additions.GRAPHICAL ABSTRACT

Wines in the northwest region of China often suffer from poor color stability, low wine acidity, and inelegant aroma description. The Lachancea thermotolerans yeasts with a high yield of lactic acid and pleasant aroma compounds can ameliorate such wines. In this study, the performances of native L. thermotolerans CVE-LT1 co-inoculated with Saccharomyces cerevisiae in different ratios and strategies (simultaneous and sequential mode) were evaluated in pilot scale fermenter (60 L) and industrial scale fermenter (520 hL) of Cabernet Sauvignon grape must with high sugar/low acidity. Results evidenced that the mixed culture of L. thermotolerans and S. cerevisiae can significantly increase lactic acid level (up to 6.98 g/L) and decrease wine pH (up to 3.57). Meanwhile, the color parameters of a* and C* in mixed culture wines were improved. The higher concentrations of volatile compounds were mainly observed in sequential inoculation treatments, including the enhancement of ethyl lactate and 2-phenylethanol. As for phenolic compounds, L. thermotolerans promoted the formation of anthocyanins derivatives and phenolic acids in pilot and industrial fermentation. In conclusion, these results indicated that L. thermotolerans LT1 can be used in hot winery regions to improve the acidity, color indexes, phenolic compounds, and aroma quality of wines.

Our current research aimed to investigate the impacts of the use of indigenous yeast Saccharomyces cerevisiae with different amounts of Lachancea thermotolerans yeasts in mixed culture on the general composition, aroma compounds, sensory analysis and yeast growth of cv. Emir wine. The utilization of L. thermotolerans in mixed cultures reduced the total acidity of wines from 5.40 to 5.19 g/L (as tartaric acid). The acidity of high acid grapes musts obtained from various viticulture areas can be relatively decreased in wine production. In addition, there may be a slight decrease in the amount of ethyl alcohol. On the other hand, increasing the inoculum level of L. thermotolerans led to an increase in the amount of higher alcohols. However, the concentration of esters declined with the higher inoculum levels. According to the sensory evaluation, the most preferred wine was the one obtained with co-inoculation of S. cerevisiae and L. thermotolerans strains at the level of 5×106 and 1×108 cells/mL, respectively. As a result, it can be said that the use of L. thermotolerans yeast in different inoculum levels has a positive effect on wine fermentation.

With global climate warming, grape berries ripen too fast in warm growing regions especially in grape growing regions in western China, which in turn results in low acidity in grapes and ultimately will cause a great negative impact on the taste and quality of wine. Lachancea thermotolerans (LT) is a non-Saccharomyces yeast that can produce a high yield of lactic acid. The mixed inoculation of LT and Saccharomyces cerevisiae can effectively increase the acidity and reduce the pH of wine, thereby maintaining the microbial and color stability. In this paper, recent progress in the research and application of LT in wine fermentation is reviewed with respect to the basic physiological, biochemical and enological characteristics of LT. Furthermore, the influence of LT on the major metabolites, aroma and color of wine and the molecular mechanism of over-production of lactic acid by LT are reviewed. Finally, the commercial application of LT is outlined. This review is expected to provide a reference for further research on LT to promote its industrial application in wine production.

In this study, three commercial non-Saccharomyces yeasts, i.e., Torulaspora delbrueckii, Metschnikowia pulcherrima and Lachancea thermotolerans were tested in combination with Saccharomyces cerevisiae to produce Italian Riesling white wines, with the fermentation progress, physical and chemical indexes of white wine, volatile profile and aromatic property of resulting white wines determined and compared, in order to elucidate their influences on the aroma quality of white wines. Results showed that, all fermentations were completed successfully, but the growth of M. pulcherrima was more easily inhibited by the proliferation of S. cerevisiae. Moreover, the ethanol concentrations of white wines from mixed fermentations were lower than that of S. cerevisiae single inoculation. Volatile compounds were detected by HS-GC-IMS, and a total of 34 volatiles were obtained. Wines inoculated with M. pulcherrima contained the highest signal intensities of ethyl caproate, ethyl isovalerate, ethyl 2-methylbutyrate, hexyl acetate, γ-nonolactone, δ-octanolactone, hexanol, 2,3-butanedione and linalool, the overall aromatic property was greatly enhanced. Wines conducted by mixed fermentation using T. delbrueckii/S. cerevisiae and L. thermotolerans/S. cerevisiae contained higher intensities of ethyl octanoate, propyl acetate, ethyl propionate and α-terpinene, and the aroma and sensory characteristics of these two pairs were similar, both better than the wine fermented singly by S. cerevisiae. Moreover, principal component analysis could better distinguish the wines produced by different mixed fermentation processes. In conclusion, this study showed that the mixed fermentation using non-Saccharomyces yeasts/S. cerevisiae pairs played an important role in the improvement of the aroma of white wine.

Increasingly high interest in yeast–yeast interactions in mixed-culture fermentation is seen along with beer consumers’ demands driving both market growth and requests for biotechnological solutions that can provide better sensory characteristics. In this study, Lachancea thermotolerans and Saccharomyces cerevisiae with a cell population ratio of 10:1 were inoculated for sour beer fermentation while the process conditions within the brewing industry remained unchanged. With L. thermotolerans producing lactic acid (1.5–1.8 g/L) and bringing down the pH to 3.3–3.4 whilst adding no foreign flavors herein, this study revealed a new natural, fruity sour beer with a soft, sour taste. In this study, the double-yeast mixed-culture fermentation produced more flavor substances than a single-culture process, and plenty of isobutyl acetate and isoamyl acetate enhanced the fruit aroma and balanced the sour beer with a refreshing taste. While playing a positive role in improving the beer’s quality, the double-yeast mixed-culture fermentation developed in this study helps to offer an alternative mass production solution for producing sour beer with the processes better controlled and the fermentation time reduced. The stress responses of the L. thermotolerans during the fermentation were revealed by integrating RNA sequencing (RNA-Seq) and metabolite data. Given that the metabolic flux distribution of the S. cerevisiae during the fermentation differed from that of the non-Saccharomyces yeasts, transcriptional analysis of non-Saccharomyces yeast and S. cerevisiae could be suitable in helping to develop strategies to modulate the transcriptional responses of specific genes that are associated with the aroma compounds released by S. cerevisiae and non-Saccharomyces yeasts. In the case of some non-Saccharomyces yeast species/strains, the diversion of alcoholic fermentation and the formation of a great number of secondary compounds may, in part, account for the low ethanol yield.

Chitosan exerts a significant influence on various chemical parameters affecting the quality of wine produced using multiple strains of Lachancea thermotolerans. The impact of chitosan on these parameters varies depending on the specific strain studied. We observed that, under the influence of chitosan, the fermentation kinetics accelerated for all examined strains. The formation of lactic acid increased by 41% to 97% across the studied L. thermotolerans strains, depending on the specific strain. This effect also influenced acidity-related parameters such as total acidity, which increased by 28% to 60%, and pH, which experienced a decrease of over 0.5 units. The consumption of malic acid increased by 9% to 20% depending on the specific strain of L. thermotolerans. Nitrogen consumption also rose, as evidenced by all L. thermotolerans strains exhibiting a residual value of Primary Amino Nitrogen (PAN) of below the detection limit, and ammonia consumption increased by 90% to 100%, depending on the strain studied. However, certain parameters such as acetic acid, succinic acid, and glycerol showed contradictory results depending on the strain under investigation. In terms of volatile composition, chitosan supplementation led to increased production of i-butanol by 32% to 65%, 3-methylbutanol by 33% to 63%, and lactic acid ethyl ester by 58% to 91% across all studied strains of L. thermotolerans. Other analyzed aroma compounds exhibited varying changes depending on the specific strain of L. thermotolerans.

The interest in and growth of craft beer has led to an intense search for new beers and styles. The revival of traditional styles has sometimes been hampered by the use of microorganisms such as lactic acid bacteria. Therefore, studies on alternative yeasts for the production of this style of beer have increased. In this work and together with previous studies carried out with yeasts isolated from Madrid agriculture (from grapes, must, wine, vineyards and wineries), the capacity of 10 yeast strains, belonging to the genus Lachancea thermotolerans, for the production of sour beer has been determined. For this purpose, different fermentation scale-ups (100 mL, 1 L and 100 L) have been performed and their fermentation capacity, aroma compound production (33 volatile compounds by GC), organoleptic profile (trained tasting panel and consumers), melatonin production (HPLC) and antioxidant capacity have been studied. Beer fermented with yeast strain CLI 1232 showed a balanced acidity with a fruity aromatic profile and honey notes. On the other hand, the beer fermented with strain 1-8B also showed a balanced acidity, but less fruity and citric flavour than CLI 1232 strain. Finally, the yeast strain selected by the consumers (CLI 1232) was used for beer production at industrial scale and the market launch of a sour beer. [ABSTRACT FROM AUTHOR]

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Non-Saccharomyces (NS) yeasts are gaining popularity in modern winemaking for improving wine quality. Climate change is one of the biggest challenges winegrowing now faces in warm regions. Here, Lachancea thermotolerans LtS1 and Torulaspora delbrueckii TdS6 combined with Saccharomyces cerevisiae ScS13 isolated from Assyrtiko grapes from Santorini island were evaluated in grape must fermentation with the aim to mitigate major consequences of temperature rise. Different inoculation protocols were evaluated, including simultaneous and sequential mixed-strain inoculations, displaying significant variation in the chemical and kinetic characteristics. Both LtS1 and TdS6 could raise the titratable acidity (TA). TdS6 also reduced the volatile acidity (VA) and was thus chosen for further evaluation in microvinifications and pilot-scale fermentations. Consistent with lab-scale trials, sequential inoculation exhibited the longest persistence of TdS6 resulting in minimum VA levels. Diethyl succinate, ethyl propanoate, and ethyl isobutyrate were significantly increased in sequential inoculations, although a decline in the net total ester content was observed. On the other hand, significantly higher levels of TA, succinic acid, and 2-methylpropanoic were associated with sequential inoculation. The overall performance of TdS6 coupled with a high compatibility with S. cerevisiae suggests its use in the fermentation of Santorini-Assyrtiko or other high sugar musts for the production of structured dry or sweet wines. [ABSTRACT FROM AUTHOR]

This work presents the attempt to develop a production technology for grape–plum low-alcohol beverages and enhance their chemical composition and flavor complexity through the non-Saccharomyces species. Saccharomyces cerevisiae (SC) pure cultures were used as reference beverages. Pure cultures of Lachancea thermotolerans (LT) and co-inoculated Lachancea thermotolerans with Saccharomyces cerevisiae (MIX) were included for grape–plum must fermentation at a pilot scale. The process involves two steps: a primary alcoholic fermentation in stainless steel tanks (F1) and a secondary fermentation in a bottle after dextrose syrup addition (F2). The chemical compositions of all beverages obtained in F1 and F2 were studied. Compared to SC, must inoculated with L. thermotolerans (LT and MIX) required four more days to complete the fermentation of sugars during F1. SC fermentation tended to have slightly higher pH and titratable acidity values and lower concentrations of total phenols. Final levels of aromatic precursor nitrogen and sulfur amino nitrogen were obtained more in SC than in LT and MIX. SC treatment had higher final levels of histidine, phenylalanine, isoleucine, lysine, methionine, threonine, valine, and cysteine. Related to individual amino acids, SC treatment had higher final levels of histidine, phenylalanine, isoleucine, lysine, methionine, threonine, valine, and cysteine. Analysis of the volatile composition showed that, compared with SC, MIX had the highest percentage of higher alcohols (3-methyl-1-butanol and 2-methyl-1-butanol) and acetates (isoamyl acetate and isobutyl acetate) which are associated with fruity and banana aromas. A decreasing trend in volatile fatty acids was observed in LT and MIX compared to SC. LT application, both in pure and mixed culture, significantly modified the values of the percentage of 5 of the 10 ethyl ester compounds analyzed. Finally, the sensory analysis showed that there were no significant differences, even though the non-Saccharomyces had a higher percentage of volatile metabolites. The results have shown that through this process an innovative and high-quality product was obtained: a low-alcohol beverage made from grapes and plums, which could be developed at an industrial level due to the increasing interest of consumers in this type of product. [ABSTRACT FROM AUTHOR]

Alcoholic beverages and vinegar products were produced using the pomace of a Finnish apple cultivar, and the influence of yeast strains on the non-volatile and volatile compound contents were studied. Apple pomace mashes were fermented with Saccharomyces cerevisiae, Torulaspora delbrueckii , and sequential inoculation with Lachancea thermotolerans and S. cerevisiae to produce alcoholic beverages. Commercial non-pasteurized vinegar was used as a starter to ferment alcoholic beverages into vinegars. Ethanol, acetic acid, sugars and organic acids were determined using GC-FID. A total of 63 volatile compounds were detected in the samples using HS-SPME-GC-MS. Alcoholic beverages fermented with the sequential inoculation had the highest total content of organic acids, and pure inoculation with T. delbrueckii resulted in the highest total content of sugars. Selection of different yeast strains did not alter the content of ethanol or acetic acid, but it significantly influenced the composition of volatile compounds in alcoholic beverages and vinegars. Compared to initial apple pomace mash, significant increases in the contents of ethyl esters (up to 4810-folds), higher alcohols (up to 24-folds) and volatile acids (up to 1853-folds) were observed in fermented products depending on the yeasts used. The main volatile compounds in vinegars prepared from S. cerevisiae fermented beverage were 2-phenylethanol, octanoic acid, and decanoic acid, from T. delbrueckii beverage 2-methylpropanoic acid and 3-methylbutanoic acid, and from sequential fermentation ethyl acetate, and ethyl 2-hydroxypropanoate. In conclusion, non- Saccharomyces yeasts have potential to produce more complex aromatic profile to fermented products derived from by-products of apple juice processing. • First research focusing on valorizing apple pomace into vinegars. • Acetic fermentation increased volatile acid contents but decreased higher alcohols. • Sequential fermentation increased complexity of volatile compound profile of vinegar. [ABSTRACT FROM AUTHOR]

This review investigates the roles of Saccharomyces sp. And non-Saccharomyces yeasts, lactic acid bacteria (LAB), and bioprotection strategies in promoting sustainable winemaking. While Saccharomyces cerevisiae has long been central to fermentation, non-Saccharomyces species like Starmerella bacillaris , Torulaspora delbrueckii , and Lachancea thermotolerans are now recognized for enhancing flavor complexity and reducing the need for chemical additives. LAB, particularly Oenococcus oeni , contribute to malolactic fermentation, improving wine stability and sensory qualities, while aiding in sustainable waste management by converting by-products into valuable materials such as biogas. Bioprotection methods using yeasts like Metschnikowia pulcherrima offer a natural alternative to chemical preservatives, reducing sulfite use and chemical inputs. These microbial strategies align with sustainability goals by minimizing synthetic additives, promoting natural fermentation, and enhancing energy efficiency. Sustainable vineyard practices, such as promoting microbial diversity, cover cropping, and organic pest management, help maintain soil health, reduce chemical fertilizers, and improve vine resilience. The integration of these practices supports both environmental and economic sustainability, reducing production costs and enhancing product quality. Additionally, advances in omics approaches enable the development of tailored microbial consortia suited to specific environmental conditions, further improving the resilience and efficiency of winemaking, especially under climate variability. This comprehensive approach meets consumer demand for natural wines while reducing the wine industry's environmental footprint and improving economic viability. [Display omitted] [ABSTRACT FROM AUTHOR]

This study evaluated effects of watermelon fermentation using different non- Saccharomyces yeast strains (Brettanomyces lambicus , Brettanomyces bruxellensis , and Lachancea thermotolerans) on the chemical profile of the derived wines. A control product was produced using Saccharomyces cerevisiae. Analytical techniques, namely HPLC-DAD-RID and LLE-GC/MS, were used to evaluate sugars, organic acids, alcohols, phenolic compounds, amino acids, and volatile compounds profiles. Fermentation reduced the sugar (glucose and fructose) and amino acid contents and increased alcohol (ethanol and glycerol), organic acids (lactic and acetic acids), phenolics, and volatile compounds. The use of the B. bruxellensis strain resulted in an alcoholic beverage with quality parameters similar to that from S. cerevisiae but with higher concentrations of phenolics (rutin, gallic acid, caftaric acid, and caffeic acid) and important volatile compounds (isoamyl acetate). The watermelon wine had a higher content of bioaccessible phenolic compounds than the must, especially catechin, procyanidin B1, procyanidin B2 and gallic acid, due to their high bioaccessibility (>70%). The yeast type influenced the chemical profile of the beverages. B. bruxellensis is a promising tool to obtain watermelon alcoholic beverage in an industrial scale. [Display omitted] • Nonconventional yeasts displayed suitable fermentation profiles for watermelon wine. • Brettanomyces and Lachancea yeasts exhibited suitable fermentation metabolites. • Fermenting B. bruxellensis produced a wine similar to that produced by S. cerevisiae. • The fermented watermelon had a higher content of bioaccessible phenolics. [ABSTRACT FROM AUTHOR]

One of the most notable effects of climate change, especially in warm regions, is the decrease in acidity (i.e., increase in pH) of wines and a reduction in their aromatic profile. To address this issue, must from a white grape variety with low acidity were inoculated with two non-Saccharomyces yeasts (Lachancea thermotolerans and Torulaspora delbrueckii) to enhance the acidity of the resulting wines. Basic oenological variables and major volatile compounds and polyols of the wines were analyzed, and the results were compared with those obtained through a Saccharomyces cerevisiae strain. Through multiple regression analysis, we found relations between the production of lactic acid to compounds involved in yeast metabolism and redox balance, including glycerol, acetic acid, isobutanol, isoamyl alcohols, and 2-phenylethanol. By means of principal component analysis, we obtained three components that explain more than 89% of the observed variability. The first component differentiates wines produced by L. thermotolerans; the second differentiates wines obtained by S. cerevisiae from those obtained by T. delbrueckii; and the third component is related to the temperature of fermentation. Organoleptic wines produced with S. cerevisiae were the best valuated, but taste was a highlight of the wines produced with L. thermotolerans due to possessing the best acidity.

Spent coffee grounds (SCG) disposal is an environmental problem. These residues from coffee brewing and instant coffee production have potential to produce novel alcoholic beverages. SCG valorization through sequential alcoholic and malolactic fermentation was investigated using a yeast, Lachancea thermotolerans Concerto and a lactic acid bacterium (LAB), Oenococcus oeni Lalvin 31 in this study. Our results showed that sequential inoculation prevented early death of yeast confronted when simultaneous inoculation was adopted, allowing for growth and persistence of both yeast and LAB till the end of fermentation. Adequate ethanol production (4.91 ± 0.13 %, v/v) with low residual sugar content was also attained. In addition, relatively lower levels of acetic, lactic, and succinic acids were produced by sequential inoculation than that of simultaneous inoculation. Furthermore, SCG hydrolysates fermented via sequential inoculation had the widest variety of volatiles (e.g. esters and ketones). Overall, our results indicated that sequential inoculation of L. thermotolerans and O. oeni in SCG hydrolysates might be a way to develop novel beverages with pleasant flavor profiles.

Global warming poses a threat to winemaking worldwide, especially in dry–warm regions such as Southern Italy. Must fermentation with non-Saccharomyces yeast starter is a possible approach to limit the negative effects of climate change, leading to desirable effects such as an increase in total acidity and/or aroma improvement. The aim of this study was to evaluate the effects of the use of a non-Saccharomyces starter (Lachancea thermotolerans) on the chemical and sensory properties of wines obtained by the the fermentation of two autochthonous Apulian grape cultivars, namely Bombino nero and Minutolo, as compared to the traditional Saccharomyces cerevisiae-driven fermentation. Bombino and Minutolo wines fermented with either Lachancea thermotolerans or Saccharomyces cerevisiae were characterized for their oenological parameters, volatile profiles, and sensory properties. Both chemical and sensory properties were affected by the yeast starter. Inoculation of L. thermotolerans increased sensory complexity, with different floral and sweet-like attributes for both cultivars. Bombino nero, a neutral cultivar, showed a clear effect on wine composition, with both an increase in lactic acid and a change in the volatile profile. On the contrary, the impact of L. thermotolerans was partially masked in Minutolo due to the strong primary aroma background of this highly terpenic cultivar. In this work, we evidenced a notable cultivar × yeast interaction, showing how generalizations of the effects of non-Saccharomyces yeasts on vinification are difficult to achieve, as they show a cultivar-specific outcome. [ABSTRACT FROM AUTHOR]

The emerging low acidity in icewine grapes is becoming a major problem in producing quality icewine. Using non-Saccharomyces cerevisiae yeasts in fermentation can improve wine's organoleptic characteristics and aromatic quality. This study evaluated two indigenous non-Saccharomyces cerevisiae yeasts, Lachancea thermotolerans (LT-2) and Torulaspora delbrueckii (TD-3), for their ability to improve the acidity and quality of 'Semillon' icewine. Five different inoculation schemes were implemented, including a single inoculation of S. cerevisiae (SC), L. thermotolerans (LT), and T. delbrueckii (TD); the sequential inoculation of L. thermotolerans, followed by S. cerevisiae after 6 days (L-S); and the sequential inoculation of L. thermotolerans, followed by T. delbrueckii after 6 days (L-D). The results showed that, during sequential fermentation (L-S and L-D), the presence of S. cerevisiae or T. delbrueckii slightly restrained the growth of L. thermotolerans. Single or sequential inoculation with L. thermotolerans and T. delbrueckii significantly reduced the amount of volatile acidity and increased the glycerol content. Furthermore, fermentations involving L. thermotolerans produced relevant amounts of lactic acid (2.04–2.2 g/L) without excessive deacidification of the icewines. Additionally, sequential fermentations increased the concentration of terpenes, C13-norisoprenoid compounds, and phenethyl compounds. A sensory analysis also revealed that sequentially fermented icewines (L-S and L-D) had more fruity and floral odors and aroma intensity. This study highlights the potential application of L. thermotolerans and T. delbrueckii in sequential fermentation to improve the icewine quality. [ABSTRACT FROM AUTHOR]

One of the alternatives to SO2 as an antimicrobial is the use of bioprotection yeasts, which colonize the medium preventing the proliferation of undesirable microorganisms. In this work, the bioprotective effect of a mixed inoculum formed by Torulaspora delbrueckii/Lachancea thermotolerans during fermentation was evaluated. For this purpose, fermentations were carried out using this mixed inoculum and the populations of yeasts, lactic bacteria and acetic bacteria, and the physical–chemical parameters of the wines obtained were studied. The results were compared with those obtained in spontaneous fermentation with and without SO2. The different fermentation strategies caused a differentiation in the yeast species present during fermentation. Regarding populations of lactic acid bacteria, results showed that the effect of the addition of the mixed inoculum was comparable to that exerted by SO2. On the other hand, due to the high sensitivity of acetic acid bacteria to SO2, the sulfite vinifications showed a lower population of acetic acid bacteria in the early stages of fermentation, followed by the vinifications with the mixed inoculum. [ABSTRACT FROM AUTHOR]

[Display omitted] • L. thermotolerans yeast strains from manna present optimal brewing characteristics. • MNF105 produced moderate amount of lactic acid (0.47 g/L). • MNF105 showed improvement of aromatic perception. • The overall organoleptic investigation showed a preference for MNF105. • MNF105 is able to produce sour beer with innovative flavours. The craft beer industry is becoming increasingly interested in the production of innovative beers. A novel approach, designated as "primary souring," employs diverse yeast species, including Lachancea thermotolerans , to produce sour beers. Furthermore, there is a growing interest in utilising unconventional yeasts to produce beers with distinctive flavours. For the first time, yeast strains of L. thermotolerans , isolated from sugar extracts of manna ash, were evaluated for their ability to produce and improve the sensory properties of sour beers. In particular, five strains exhibited notable resistance to ethanol, sugar and hops, as well as comparable lactic acid production (ranging from 0.33 to 0.45 g/L). Experimental beers produced using MNF105 (T1) were perceived as the most "fruity". This is the first study to examine the impact of this novel indigenous strain, derived from unconventional matrixes such as manna, on the organoleptic quality of craft sour beers. Consequently, elevated levels of ethyl decanoate, ethyl hexanoate, ethyl octanoate and ethyl nonanoate were found in T1 beer, exceeding the perception threshold. The ability of this strain to perform light bio-acidification is a valuable feature for the development of new brewing techniques, particularly for the creation of sour beers with balanced acidity and innovative flavours. The yeast L. thermotolerans MNF105, which is related to manna, has excellent technological properties and is a promising starter for beer production with the ability to light bio-acidify and modulate flavour. [ABSTRACT FROM AUTHOR]

Lack of acidity in wine is one of the main challenges in warm wine-producing regions. The non- Saccharomyces yeast, Lachancea thermotolerans (LT), is well known for its great potential in wine acidification through the production of lactic acid. In this study, an indigenous LT strain was sequentially inoculated with S. cerevisiae (SC). The influence of the timing of SC inoculation on wine quality was extensively studied. Inoculating SC 72 h after the inoculation of LT delayed the peak of fermentation rate, but had no significant influence on the duration of fermentation. The late inoculation (72 h) of SC effectively increased the production of lactic acid, which consequently increased the total wine acidity and decreased the pH. Higher levels of esters were also achieved by the late inoculation. Pilot-scale fermentations were carried out with both Chardonnay and Cabernet Sauvignon to verify the effect of late inoculation. Lactic acid produced in sequential fermentation was 2.73 g/L for Chardonnay wine and 5.35 g/L for Cabernet Sauvignon wine, and the production of ethyl esters was also enhanced. Overall, the results demonstrated that the late inoculation of SC in sequential fermentation with LT improved wine quality by increasing acidity and producing more balanced and aromatic wines. • SC inoculation timing in sequential fermentation with LT significantly affected wine quality. • Late inoculation of SC markedly increased lactic acid production. • Higher levels of esters were achieved with late SC inoculation. • The quality of Chardonnay and Cabernet Sauvignon wines was improved by late inoculation of SC. [ABSTRACT FROM AUTHOR]

Viognier is a warm climate grape variety prone to loss of acidity and accumulation of excessive sugars. The yeast Lachancea thermotolerans can improve the stability and balance of such wines due to the partial conversion of sugars to lactic acid during alcoholic fermentation. This study compared the performance of five L. thermotolerans strains in co-inoculations and sequential inoculations with Saccharomyces cerevisiae in high sugar/pH Viognier fermentations. The results highlighted the dichotomy between the non-acidified and the bio-acidified L. thermotolerans treatments, with either comparable or up to 0.5 units lower pH relative to the S. cerevisiae control. Significant differences were detected in a range of flavour-active yeast volatile metabolites. The perceived acidity mirrored the modulations in wine pH/TA, as confirmed via "Rate-All-That-Apply" sensory analysis. Despite major variations in the volatile composition and acidity alike, the varietal aromatic expression (i.e., stone fruit aroma/flavour) remained conserved between the treatments. [ABSTRACT FROM AUTHOR]

In wine fermentation, improved wine complexity and sensorial properties can arise from the use of non-Saccharomyces yeast. Generally less alcohol tolerant, such strains often do not finish fermentation, therefore requiring a second inoculation with the more robust Saccharomyces cerevisiae, usually added on Day 3. This sequential approach affords non-Saccharomyces time to make an impact before being overtaken by S. cerevisiae. However, two inoculations are inconvenient; therefore the identification of a slow growing S. cerevisiae strain that can be used in a single co-inoculation with the non-Saccharomyces yeast is highly attractive. In this study we investigated the use of the naturally occurring ‘loss of function’ SER1 variant, identified in a Sake yeast, for the purposes of carrying out co-inoculated wine fermentations. The SER1-232(G > C; G78R) change was introduced into the commonly used wine strain, EC1118, via CRISPR/Cas9 editing. In a chemically defined grape juice medium, the SER1(G78R) mutant grew and fermented more slowly and increased acetic acid, succinic acid and glycerol concentrations. Simultaneous inoculation with the slower-growing mutant with a Metschnikowia pulcherrima or Lachancea thermotolerans strain in sterile Sauvignon blanc juice resulted in differences in sensorial compounds, most likely derived from the presence of non-Saccharomyces yeasts. The EC1118 SER1 (G78R) mutant completed fermentation with M. pulcherrima, MP2, and in fact improved the viability of MP2 compared to when it was used as a monoculture. The SER1 (G78R) mutant also promoted both the growth of the SO2-sensitive L. thermotolerans strain, Viniflora® Concerto™, in a juice high in SO2 and its subsequent dominance during fermentation. In co-fermentations with wild-type EC1118, the Concerto™ population was substantially reduced with no significant changes in wine properties. This research adds to our understanding of the use of a novel slow-growing S. cerevisiae yeast in wine fermentations co-inoculated with non-Saccharomyces strains.

The interest in and growth of craft beer has led to an intense search for new beers and styles. The revival of traditional styles has sometimes been hampered by the use of microorganisms such as lactic acid bacteria. Therefore, studies on alternative yeasts for the production of this style of beer have increased. In this work and together with previous studies carried out with yeasts isolated from Madrid agriculture (from grapes, must, wine, vineyards and wineries), the capacity of 10 yeast strains, belonging to the genus Lachancea thermotolerans, for the production of sour beer has been determined. For this purpose, different fermentation scale-ups (100 mL, 1 L and 100 L) have been performed and their fermentation capacity, aroma compound production (33 volatile compounds by GC), organoleptic profile (trained tasting panel and consumers), melatonin production (HPLC) and antioxidant capacity have been studied. Beer fermented with yeast strain CLI 1232 showed a balanced acidity with a fruity aromatic profile and honey notes. On the other hand, the beer fermented with strain 1-8B also showed a balanced acidity, but less fruity and citric flavour than CLI 1232 strain. Finally, the yeast strain selected by the consumers (CLI 1232) was used for beer production at industrial scale and the market launch of a sour beer.

Background: The vineyard is a great reservoir of autochthonous yeast strains whose composition is defined by different regional (edaphology, orography or climatology) and anthropological factors (cultivation systems or cultural practices). Most of this yeast diversity corresponds to non-Saccharomyces strains, some of which have potential use in winemaking. Methods: The oenological potential of 29 different native non-Saccharomyces strains belonging to 4 species (Lachancea thermotolerans, Torulaspora delbrueckii, Starmerella bacillaris and Metschnikowia spp.) was evaluated, using the autochthonous Saccharomyces cerevisiae XG3 strain as a control. Microfermentations with pure culture of each strain were performed in duplicate and the basic parameters and major volatiles of wines were analysed following official methodology. The best strain within each species was selected using a quantification matrix including the relevant oenological characteristics. Results: The fermentative ability of non-Saccharomyces was lower than S. cerevisiae in all cases, but with differences among species. L. thermotolerans and T. delbrueckii showed higher fermentation rates than Starm. bacillaris, whereas Metschnikowia spp. presented a low fermentative power. At chemical level all non-Saccharomyces strains reduced the alcoholic content, the higher alcohols and the volatile acidity of wines and increased the content of glycerol, with differences among strains within a given species. T. delbrueckii and L. thermotolerans increased the total acidity of wines. The latter and Metschnikowia spp. strains produced lactic acid, which decreased the wine pH in the case of L. thermotolerans. According to their oenological traits the best rated strains of each species were Lt93, Td315, Mf278 and Sb474. In addition, the data obtained in pure fermentations were correlated to those chemical and aromatic compounds obtained with these non-Saccharomyces strains in sequential fermentations. Conclusions: Autochthonous strains of non-Saccharomyces yeast species contribute distinctive chemical characteristics to the wines. The correlations observed between wines fermented with the different non-Saccharomyces indigenous strains in pure and sequential fermentations suggest that their contribution to wine properties remains stable regardless of must composition or winemaking techniques.

Non-Saccharomyces (NS) yeasts are gaining popularity in modern winemaking for improving wine quality. Climate change is one of the biggest challenges winegrowing now faces in warm regions. Here, Lachancea thermotolerans LtS1 and Torulaspora delbrueckii TdS6 combined with Saccharomyces cerevisiae ScS13 isolated from Assyrtiko grapes from Santorini island were evaluated in grape must fermentation with the aim to mitigate major consequences of temperature rise. Different inoculation protocols were evaluated, including simultaneous and sequential mixed-strain inoculations, displaying significant variation in the chemical and kinetic characteristics. Both LtS1 and TdS6 could raise the titratable acidity (TA). TdS6 also reduced the volatile acidity (VA) and was thus chosen for further evaluation in microvinifications and pilot-scale fermentations. Consistent with lab-scale trials, sequential inoculation exhibited the longest persistence of TdS6 resulting in minimum VA levels. Diethyl succinate, ethyl propanoate, and ethyl isobutyrate were significantly increased in sequential inoculations, although a decline in the net total ester content was observed. On the other hand, significantly higher levels of TA, succinic acid, and 2-methylpropanoic were associated with sequential inoculation. The overall performance of TdS6 coupled with a high compatibility with S. cerevisiae suggests its use in the fermentation of Santorini-Assyrtiko or other high sugar musts for the production of structured dry or sweet wines.

This work presents the attempt to develop a production technology for grape–plum low-alcohol beverages and enhance their chemical composition and flavor complexity through the non-Saccharomyces species. Saccharomyces cerevisiae (SC) pure cultures were used as reference beverages. Pure cultures of Lachancea thermotolerans (LT) and co-inoculated Lachancea thermotolerans with Saccharomyces cerevisiae (MIX) were included for grape–plum must fermentation at a pilot scale. The process involves two steps: a primary alcoholic fermentation in stainless steel tanks (F1) and a secondary fermentation in a bottle after dextrose syrup addition (F2). The chemical compositions of all beverages obtained in F1 and F2 were studied. Compared to SC, must inoculated with L. thermotolerans (LT and MIX) required four more days to complete the fermentation of sugars during F1. SC fermentation tended to have slightly higher pH and titratable acidity values and lower concentrations of total phenols. Final levels of aromatic precursor nitrogen and sulfur amino nitrogen were obtained more in SC than in LT and MIX. SC treatment had higher final levels of histidine, phenylalanine, isoleucine, lysine, methionine, threonine, valine, and cysteine. Related to individual amino acids, SC treatment had higher final levels of histidine, phenylalanine, isoleucine, lysine, methionine, threonine, valine, and cysteine. Analysis of the volatile composition showed that, compared with SC, MIX had the highest percentage of higher alcohols (3-methyl-1-butanol and 2-methyl-1-butanol) and acetates (isoamyl acetate and isobutyl acetate) which are associated with fruity and banana aromas. A decreasing trend in volatile fatty acids was observed in LT and MIX compared to SC. LT application, both in pure and mixed culture, significantly modified the values of the percentage of 5 of the 10 ethyl ester compounds analyzed. Finally, the sensory analysis showed that there were no significant differences, even though the non-Saccharomyces had a higher percentage of volatile metabolites. The results have shown that through this process an innovative and high-quality product was obtained: a low-alcohol beverage made from grapes and plums, which could be developed at an industrial level due to the increasing interest of consumers in this type of product.

Global warming poses a threat to winemaking worldwide, especially in dry–warm regions such as Southern Italy. Must fermentation with non-Saccharomyces yeast starter is a possible approach to limit the negative effects of climate change, leading to desirable effects such as an increase in total acidity and/or aroma improvement. The aim of this study was to evaluate the effects of the use of a non-Saccharomyces starter (Lachancea thermotolerans) on the chemical and sensory properties of wines obtained by the the fermentation of two autochthonous Apulian grape cultivars, namely Bombino nero and Minutolo, as compared to the traditional Saccharomyces cerevisiae-driven fermentation. Bombino and Minutolo wines fermented with either Lachancea thermotolerans or Saccharomyces cerevisiae were characterized for their oenological parameters, volatile profiles, and sensory properties. Both chemical and sensory properties were affected by the yeast starter. Inoculation of L. thermotolerans increased sensory complexity, with different floral and sweet-like attributes for both cultivars. Bombino nero, a neutral cultivar, showed a clear effect on wine composition, with both an increase in lactic acid and a change in the volatile profile. On the contrary, the impact of L. thermotolerans was partially masked in Minutolo due to the strong primary aroma background of this highly terpenic cultivar. In this work, we evidenced a notable cultivar × yeast interaction, showing how generalizations of the effects of non-Saccharomyces yeasts on vinification are difficult to achieve, as they show a cultivar-specific outcome.

Climate change is affecting vineyards, resulting in grapes with a low acidity a high pH and sugar at harvest time. The most common procedure so far to improve the acidity and reduce the final pH of wines is to use tartaric acid, but wine can also be acidified microbiologically using Lachancea thermotolerans yeasts, a natural bio-tool that acidifies gradually during the first stage/days of fermentation. Two strains of L. thermotolerans were compared with one Saccharomyces cerevisiae at a pilot-scale under similar fermentation conditions and in duplicate. A sequential inoculation was performed on the third day for the non-Saccharomyces, producing only about 1 g/L of lactic acid, which was suitable for comparison with the Saccharomyces, to which 1.5 g/L of tartaric acid had been added to lower the final pH. The three fermentations ended with a total acidity without significant differences. A significant and normal feature of the L. thermotolerans yeasts is their higher propane-1,2,3-triol production, which was observed in the Laktia yeast, and the acetic acid was <0.3 g/L. The amount of volatile metabolites was generally higher for non-Saccharomyces and the increase was seen in carbonyl compounds, organic acids, lactones, fumaric compounds, and phenols. Finally, the sensory analysis showed that there were hardly any significant differences, even though the non-Saccharomyces had a higher quantity of volatile metabolites, which could lead to a good acceptance of the product, since biological acidification was used, generating a more natural product. [ABSTRACT FROM AUTHOR]

Non-Saccharomyces yeasts constitute a useful tool in winemaking because they secrete hydrolytic enzymes and produce metabolites that enhance wine quality; in addition, their ability to reduce alcohol content and/or to increase acidity can help to mitigate the effects of climatic change on wines. The purpose of this study was to evaluate the oenological traits of non-Saccharomyces yeast strains autochthonous from Galicia (NW Spain). To do that, we carried out sequential fermentation using 13 different species from the yeast collection of Estación de Viticultura e Enoloxía de Galicia (Evega) and Saccharomyces cerevisiae EC1118. The fermentation kinetics and yeast implantation were monitored using conventional methods and genetic techniques, respectively. The basic chemical parameters of wine were determined using the OIV official methodology, and the fermentative aroma compounds were determined by GC–FID. The results evidenced the limited fermentative power of these yeasts and the differences in their survival after the addition of S. cerevisiae to complete fermentation. Some strains reduced the alcohol and/or increased the total acidity of the wine. The positive effect on sensory wine properties as well as the production of desirable volatile compounds were confirmed for Metschnikowia spp. (Mf278 and Mp176), Lachancea thermotolerans Lt93, and Pichia kluyveri Pkl88. These strains could be used for wine diversification in Galicia. [ABSTRACT FROM AUTHOR]

The use of non-Saccharomyces yeasts in sequential fermentations with S. cerevisiae has been proposed to improve the organoleptic characteristics involved in the quality of wine. The present study set out to select a non-Saccharomyces inoculum from the D.O.Ca. Rioja for use in winemaking. Strains included in the study belonged to Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, Zygosaccharomyces bailii, Williopsis pratensis, Debaryomyces hansenii, Pichia kluyveri, Sporidiobolus salmonicolor, Candida spp., Cryptococcus spp. and two mixed inocula of Lachancea thermotolerans-Torulaspora delbrueckii in a 30/70 ratio. In the first stage of the process, SO2 resistance and presence of enzymatic activities related to wine aroma and wine color and fining (esterase, esterase-lipase, lipase, leucine arylamidase, valine arylamidase, cystine arylamidase, β-glucosidase, pectinase, cellulose, xylanase and glucanase) were studied. In the later stages, selection criteria such as fermentative behavior, aroma compound production or influence on phenolic compounds were studied in laboratory scale vinifications. Taking into account the results obtained in the different stages of the process, a mixed inoculum of Lachancea thermotolerans-Torulaspora delbrueckii in a 30/70 ratio was finally selected. This inoculum stood out for its high implantation capacity, the production of compounds of interest such as glycerol and lactic acid and the consequent modulation of wine acidity. Given these characteristics, the selected inoculum is suitable for the production of quality wines. [ABSTRACT FROM AUTHOR]

Traditionally, the use of malolactic fermentation gives rise to microbiologically stable wines. However, malolactic fermentation is not free from possible collateral effects that can take place under specific scenarios. The present work tests the influence of different biological deacidification strategies on the volatile and non-volatile components of white must from Germany. The study compared mixed cultures of Lachancea thermotolerans and Schizosaccharomyces pombe and a pure culture of Sc. pombe to the classical biological deacidification process performed by lactic acid bacteria. Strains of Oenococcus oeni and Lactiplantibacillus plantarum were co- or sequentially inoculated with S. cerevisiae to carry out malolactic fermentation. Different fermentation treatments took place at a laboratory scale of 0.6 L in vessels of 0.75 L. The instrumental techniques Fourier-transform mid-infrared spectroscopy (FT-MIR), high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) were used to evaluate different chemical parameters in the final wines. The results showed the ability of Sc. pombe to consume malic acid in combination with L. thermotolerans without using S. cerevisiae or lactic acid bacteria. Fermentations involving Sc. pombe consumed all the malic acid, although they reduced the concentrations of higher alcohols, fatty acids and acetic acid. Simultaneous alcoholic and malolactic fermentations reduced malic acid by about 80%, while classical malolactic fermentation reduced it by 100%. Fermentations involving L. thermotolerans produced the highest lactic acid, ester and glycerol concentrations. [ABSTRACT FROM AUTHOR]

Abstract Background In wine fermentation starter cultures, the blending of non-Saccharomyces yeast with Saccharomyces cerevisiae to improve the complexity of wine has become common practice, but data regarding the impact of co-cultivation on yeast physiology and on genetic and metabolic regulation remain limited. Here we describe a transcriptomic analysis of mixed fermentations of Saccharomyces cerevisiae and Lachancea thermotolerans. The fermentations were carried out in carefully controlled environmental conditions in a bioreactor to reduce transcriptomic responses that would be due to factors other than the presence of the second species. Results The transcriptomic data revealed that both yeast species showed a clear response to the presence of the other. Affected genes primarily belonged to two groups: genes whose expression can be linked to the competition for certain trace elements such as copper and iron, as well as genes required for cell wall structure and integrity. Furthermore, the data revealed divergent transcriptional responses with regard to carbon metabolism in response to anoxic conditions. Conclusions The results suggest that the mixed fermentation created a more competitive and stressful environment for the two species than single strain fermentations independently from total biomass, i.e. competition between cells of the same species is less stressful, or may present a different set of challenges, than interspecies competition. The changes in cell wall and adhesion properties encoding genes suggest that the adjustment of physical contact between cells may play a direct role in the response to the presence of competing species.