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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.

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.

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.

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.

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 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.

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, SO 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., This research was funded by Instituto Nacional de Investigaciones Agrarias (INIA) (Project RTA2013-0053-C03).

Lactic acid production is an important feature of the yeast Lachancea thermotolerans that has gained increasing interest in winemaking. In particular, in light of climate change, the biological acidification and ethanol reduction by the use of selected yeast strains may counteract the effect of global warming in wines. Here, the enological potential of a high lactate-producing L. thermotolerans strain (P-HO1) in mixed fermentations with S. cerevisiae was examined. Among the different inoculation schemes evaluated, the most successful implantation of L. thermotolerans was accomplished by sequential inoculation of S. cerevisiae, i.e., at 1% vol. ethanol. P-HO1produced the highest levels of lactic acid ever recorded in mixed fermentations (10.4 g/L), increasing thereby the acidity and reducing ethanol by 1.6% vol. L. thermotolerans was also associated with increases in ethyl isobutyrate (strawberry aroma), free SO2, organoleptically perceived citric nuances and aftertaste. To start uncovering the molecular mechanisms of lactate biosynthesis in L. thermotolerans, the relative expressions of the three lactate dehydrogenase (LDH) paralogous genes, which encode the key enzyme for lactate biosynthesis, along with the alcohol dehydrogenase paralogs (ADHs) were determined. Present results point to the possible implication of LDH2, but not of other LDH or ADH genes, in the high production of lactic acid in certain strains at the expense of ethanol. Taken together, the important enological features of P-HO1 highlighted here, and potentially of other L. thermotolerans strains, indicate its great importance in modern winemaking, particularly in the light of the upcoming climate change and its consequences in the grape/wine system.