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Review

Biotechnological Strategies for Ethanol Reduction in Wine

by
Bruno Testa
1,
Francesca Coppola
2,*,
Mariantonietta Succi
1,* and
Massimo Iorizzo
1
1
Department of Agriculture, Environmental and Food Sciences, University of Molise, 86100 Campobasso, Italy
2
Institute of Food Sciences, National Research Council of Italy, 83100 Avellino, Italy
*
Authors to whom correspondence should be addressed.
Fermentation 2025, 11(3), 159; https://doi.org/10.3390/fermentation11030159
Submission received: 10 March 2025 / Revised: 17 March 2025 / Accepted: 19 March 2025 / Published: 20 March 2025
(This article belongs to the Special Issue Recent Advances in Microbial Fermentation in Foods and Beverages)
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Abstract

:
In recent years, wine producers have become increasingly interested in partial or total dealcoholisation of wines due to changing consumer preferences, increased attention to health aspects of alcohol consumption, and the impact of climate change on sugar concentration in grapes. Strategies for decreasing alcohol content in wines include pre-fermentation, fermentation, and post-fermentation techniques. These approaches allow for the reduction of fermentable sugars before fermentation, limit the alcohol production during fermentation, and separate alcohol after fermentation. From a biotechnological perspective, yeasts play a critical role in alcoholic fermentation, directly influencing the final alcohol content of the product. The use of non-Saccharomyces yeasts in co-inoculation or sequential inoculation with Saccharomyces yeasts represents a promising biotechnological strategy for the reduction of alcohol in wine.

1. Introduction

In recent years, the wine market has focused on beverages with a moderate alcohol content without additives or with natural additives and enriched with health-promoting components such as antioxidants [1,2,3]. Based on alcoholic content, wines can be classified as alcohol-free (<0.5% v/v), low-alcohol (0.5% to 1.2% v/v), reduced-alcohol (1.2% to 5.5% or 6.5% v/v), lower-alcohol (5.5% to 10.5% v/v), and alcoholic wines (>10.5% v/v) [4,5]. This classification, which can vary between countries, is based on labelling and different legislative regulations [6]. The focus on consumer health and well-being has led to increased demand for low- and no-alcohol wines. These beverages preserve some of the flavours and aromas of traditional wines. They also offer health benefits, particularly in terms of cardiovascular health [7]. Over the years, wine producers and researchers have developed various techniques to reduce the alcohol content of wines in order to meet the demands of consumers who want wines with lower alcohol content and, at the same time, good sensory quality [5,8].
A partial or total reduction in alcohol content can be achieved in the various stages of wine production (pre-fermentative, fermentative, and post-fermentation phases) using different strategies: viticultural practices, such as decreasing the ratio between leaf area and fruit weight in order to reduce the concentration of sugar in the berry; pre-fermentation and winemaking practices, including dilution and elimination of sugars from grape must; microbiological strategies, such as the selection and use of yeasts that have a lower efficiency for the conversion of sugars into alcohol; and post-fermentation processing practices and technologies, which involve the physical dealcoholisation (removal of ethanol) of wine [7,9,10]. In this scenario, microbiological strategies play a fundamental role in the production of wines with low alcohol content, allowing winemakers to control ethanol levels while preserving the organoleptic peculiarities of the wine (Figure 1).
This biotechnological approach mainly involves the selection of native yeast strains [11], the use of non-Saccharomyces yeasts, and controlled fermentation techniques [12,13,14,15,16]. Used as starters in winemaking, some non-Saccharomyces yeasts produce less ethanol than Saccharomyces cerevisiae [12,13], synthesising many compounds that contribute to the aromatic complexity of wines [14,15]. Another promising biotechnological approach involves the genetic modification of yeasts to alter metabolic pathways associated with ethanol production [16,17], promoting the synthesis of glycerol as an alternative to ethanol [18], and modifying the activity of pyruvate decarboxylase to redirect sugar metabolism and inhibit ethanol production [19,20]. However, regulatory constraints on genetically modified organisms (GMOs) limit the use of these yeasts in winemaking [21].
During winemaking, fermentation parameters such as temperature, oxygen availability, and nitrogen supplementation can be modified, affecting significantly yeast metabolism and ethanol production [22,23]. Low-temperature fermentations (12–15 °C) have been shown to reduce yeast metabolic efficiency, resulting in lower ethanol yields [24]. Oxygenation during the early stages of fermentation can also shift metabolic pathways towards biomass production and thus can be used as an effective and natural method to reduce the alcohol content of wine without penalising it organoleptically [25,26].
Future research into yeast biotechnology may contribute to improving the quality of low-alcohol wines and increase their acceptance in the global market. The main results obtained in studies that have applied biotechnological approaches to reduce the alcohol content of wines are discussed below.

2. Saccharomyces Yeasts and Their Hybrids

The role of yeasts is crucial in obtaining low-alcohol wines. In this field, fermentation management strategies can be adopted in order to regulate the alcohol concentration. [27,28]. S. cerevisiae is the main yeast species utilised in winemaking, characterised by its high fermentation power, effective conversion of sugar to alcohol, and high alcohol resistance [29,30]. A recent study by Tronchoni et al. [31] focused attention on the optimisation of fermentation conditions of grape must obtain a reduced ethanol yield by using selected S. cerevisiae strains suitable for aerobic fermentation. In the past, some studies have explored innovative biotechnological strategies to reduce the alcohol content of wines using engineered S. cerevisiae and non-Saccharomyces or Saccharomyces non-cerevisiae yeasts [32,33]. While studies on engineered yeasts have slowed due to consumers perception that GMOs may have some health risks [34], the use of non-Saccharomyces and Saccharomyces non-cerevisiae yeasts represents a promising alternative to regulate the alcohol content in the wine while maintaining the desired sensory characteristics [35]. Table 1 summarises the main effects of Saccharomyces non-cerevisiae yeasts on ethanol levels in wine.
Among Saccharomyces non-cerevisiae yeasts, Saccharomyces bayanus is a species that, under determined conditions, can exhibit reduced fermentation activity. This yeast is used in winemaking for the production of sparkling wines but if used under certain fermentation conditions it can help to control alcohol production [42].
In a recent study by González et al. [47], S. bayanus, co-inoculated with S. cerevisiae, was found to be able to modify metabolic pathways to generate more glycerol and organic acids, thus improving the flavour profile of low-alcohol wines.
Other Saccharomyces species, such as Saccharomyces uvarum, Saccharomyces kudriavzevii, Saccharomyces eubayanus, Saccharomyces paradoxus, and Saccharomyces mikatae, represent alternative options for winemaking of low alcohol wines.
S. uvarum is a promising yeast species recognised for its lower fermentation attitude than S. cerevisiae [48] and associated with a strong aromatic intensity of wine thanks to the high production of acetate esters [49]. It is a cryotolerant yeast that allows fermentation at low temperatures (12–20 °C), resulting in an alcohol reduction of approximately 2% (v/v) compared to the traditional S. cerevisiae yeast [42]. S. uvarum is characterised by low acetic acid production and high yields of glycerol and lactic acid. In the study by Muratore et al. [50], it was found that the characteristics of Malvasia delle Lipari wine fermented with the cryotolerant S. uvarum were better than those of the same wine fermented with S. cerevisiae. S. uvarum gave a final product with lower levels of volatile acidity and ethanol, while improving the wine sensory profile.
A significant decrease in alcohol content (1.7% v/v) was obtained by using S. uvarum in the vinification of Merlot wine. However, this process resulted in a wine with a compromised sensory profile [43]. In this regard, it has been found that S. uvarum can produce by-products such as acetic acid and higher alcohols, which could negatively affect the flavour of wine [42]. Therefore, optimisation of fermentation conditions is necessary to optimally manage this species in the production of low-alcohol wines.
S. kudriavzevii has physiological properties of particular importance in winemaking, such as its good fermentation at low temperatures, which results in wines with higher glycerol [51] and lower alcohol content [22,52]. In addition, S. kudriavzevii has been shown to produce high concentrations of higher alcohols, such as 2-phenylethanol [53]. S. eubayanus is a natural hybrid of S. pastorianus and S. bayanus [54,55,56], which is associated with the production of various alcoholic fermented beverages such as beer, cider and wine. S. eubayanus is capable of fermenting under low nitrogen concentrations and at low temperatures [57]. The sensitivity of this species to ethanol has been shown to increase with temperature [37]. In the study by Parpinello et al. [24], strains of S. eubayanus were tested on a laboratory scale for the winemaking of Chardonnay wine. The results showed that these strains were able to reduce the alcohol content by approximately 2% (v/v).
Another yeast from the Saccharomyces sensu stricto complex, S. paradoxus, has also shown promising fermentation performances. In a recent study by Costantini et al. [45], S. paradoxus ISE1618 was used for the vinification of Grignolino grapes. Compared to wine produced with S. cerevisiae, wine made with S. paradoxus was characterised by a high glycerol production and a lower alcohol content. In addition, some S. paradoxus strains had higher concentrations of chitin in the yeast cell wall. This property allows them to bind grape chitinases more efficiently than conventional S. cerevisiae strains, improving the protein stability of wine [58].
In the last few decades, S. uvarum, S. kudriavzevii and S. cerevisiae hybrids have gained popularity in wine research and industry. A study by Pérez et al. [59] showed that intra- and interspecific hybrids of these species are able to produce lower ethanol concentrations and increased amounts of glycerol, 2,3-butanediol, and organic acids.
These results support the fact that artificial hybridisation is a promising strategy to improve the performance of industrial yeast strains [60,61,62]. A study by Bellon et al. [46] described novel hybrids obtained by interspecific hybridisation between S. cerevisiae AWRI838 (an isolate of the commercial wine yeast strain EC1118) and S. mikatae NCYC2888 (designated AWRI1529), a diploid, prototrophic, heterozygous and homothallic wild yeast strain. The wines produced with these hybrids had different concentrations of volatile compounds compared to wines fermented with S. cerevisiae alone. In particular, the hybrid CxM3 produced lower concentrations of ethanol compared to the wine yeast parent S. cerevisiae AWRI838, and was also one of the largest producers of glycerol.

3. Non-Saccharomyces Yeasts

Among the various existing strategies to reduce the alcohol content of wines, the use of non-Saccharomyces yeasts has attracted increasing interest in recent years [17,63]. Table 2 summarises the main effects of non-Saccharomyces yeasts on ethanol levels in wine.
To achieve this goal, these yeasts can basically be used under anaerobic or aerobic conditions [70,73]. Aerobic fermentation, also referred to as the Crabtree effect in yeast, is a well-studied phenomenon that allows many eukaryotic cells to attain higher growth rates at high glucose availability. In this regard, Röcker et al. [71] obtained a preliminary and partial aerobic degradation of sugars by several non-Saccharomyces Crabtree-negative yeasts, before the addition of Saccharomyces yeasts for the final anaerobic fermentation. In particular, the use of Metschnikowia pulcherrima V-131 resulted in alcohol reductions of up to 3.8% (v/v).
Non-Saccharomyces yeasts produce less ethanol and are less resistant to ethanol than Saccharomyces yeasts. These characteristics are the main factors of the dominance of Saccharomyces over the other yeasts during the fermentation of wine [63]. In particular, species belonging to the genera Hanseniaspora, Candida, Pichia, Kluyveromyces, Metschnikowia, Torulospora, Starmerella, etc., which are frequently or occasionally found in grape juice, do not tolerate ethanol concentrations above 5–7% (v/v) [63,115]. The non-Saccharomyces yeasts most commonly used for alcohol reduction studies generally include the well-known wine-related species M. pulcherrima, Torulospora delbrueckii, and Starmarella bacillaris (synonym Candida zemplinina) [71,90,116]. In addition, non-Saccharomyces yeasts have been proposed in many studies to improve the sensory complexity and aroma of wine [12,90,94,116,117,118]. Furthermore, some non-Saccharomyces species exhibit antibacterial properties against spoilage bacteria and yeasts [119]. In a recent study, Testa et al. [12] showed that the strain M. pulcherrima AS3C1 exhibited antimicrobial activity against various undesirable bacteria and yeasts typically present in grape must, which can lead to wine alterations. The authors also found that M. pulcherrima AS3C1 is a low ethanol producer, around 4.0% (v/v), while exhibiting significant β-glucosidase activity. Thus, the incorporation of non-Saccharomyces yeasts into winemaking represents an attractive approach for the production of low-alcohol wines while improving the overall quality and sensory attributes of the final product [120].
Among non-Saccharomyces yeasts, S. bacillaris stands out as a species characterised by a strong fructophilic nature, the ability to withstand low temperatures and the ability to grow in high-sugar environments. Some S. bacillaris strains produce a minimal ethanol yield from consumed sugars, elevated glycerol content and moderate production of volatile acidity [121].
T. delbrueckii has recently attracted considerable attention in the wine industry for its ability to address current oenological challenges and improve wine quality compared to the use of S. cerevisiae species. T. delbrueckii is known to produce low levels of acetic acid, a key quality parameter in wine production. In addition, this yeast typically produces wines with lower ethanol concentrations compared to traditional fermentation methods [73], while increasing glycerol content [68]. This property could be a valuable asset in mitigating the challenges posed by climate change, particularly in preventing the production of high-alcohol wines associated with elevated sugar levels in grape must. T. delbrueckii shows a remarkable ability to release mannoproteins and polysaccharides during the winemaking process, which significantly improves the mouthfeel quality of the wine [122].
The genus Hanseniaspora, in particular H. uvarum, is a non-Saccharomyces yeast that is commonly found at high concentrations on the surface of grapes and during fermentation. Research by Mestre et al. [85] has shown that some strains of H. uvarum exhibit promising oenological properties, such as the ability to thrive in conditions with high sugar, ethanol and SO2 concentrations. They also produce significant amounts of glycerol while maintaining low levels of acetic acid and hydrogen sulphide, and release proteolytic enzymes. In particular, H. uvarum has the potential to produce low ethanol levels, as it requires over 19 g/L of sugar to produce 1% (v/v) ethanol [123].
Among other non-Saccharomyces yeast, Zygosaccharomyces spp. are commonly considered spoilage yeasts due to their high tolerance to osmotic and acidic stress [124]. These yeasts, naturally present in grapes and musts, can improve the production of higher alcohols during the alcoholic fermentation, while reducing the acetoin concentration. Additionally, some authors highlighted the attitude of Zygosaccharomyces bailii to produce lower ethanol concentration when used in mixed wine fermentations [40,125].
It is evident that the implementation of non-Saccharomyces yeasts, as a strategy for the production of wine with reduced alcohol content, presents interesting prospects. However, these yeasts ferment more slowly than standard Saccharomyces cultures, and therefore careful monitoring is important throughout the fermentation process in order to avoid complications such as arrested fermentations or wine alterations caused by collateral metabolic activities. Furthermore, it is important to remember that the low alcohol tolerance of these yeasts could lead to high residual sugar in wines, making their microbiological stability more difficult. [47]. Thus, the use of controlled multi-starter fermentation, using selected cultures of non-Saccharomyces and S. cerevisiae yeast strains, has been encouraged [126].

4. Genetically Engineered Saccharomyces Yeasts

Metabolic engineering approaches, which have been successfully used to optimise endogenous metabolic pathways, have been gradually replaced in recent years by evolutionary engineering strategies, aimed at generating strains with phenotypes possessing specific metabolic activities [127]. The use of genetically modified yeasts is an effective strategy for the production of low-alcohol wines. It allows ethanol content reduction without changing the organoleptic characteristics of the wine by modifying the metabolic pathways involved in fermentation. [128]. In order to produce wine with a reduced percentage of ethanol, approaches based on GMOs have focused on diverting part of the carbon flux from the ethanol pathway to other secondary products. By regulating metabolic fluxes, more carbon can be directed towards glycerol synthesis rather than ethanol production, contributing to the production of wines with lower alcohol content [129]. The overexpression of GPD1 and/or GDP2 genes, which encode isozymes of glycerol-3-phosphate dehydrogenase, reduces ethanol and increases glycerol production. In a study conducted by Cuello et al. [130], a diploid mutant of S. cerevisiae strain BY4743 pdc2Δ519 was used in a lab scale vinification leading to an ethanol reduction content by up to 7.4% (v/v), without affecting the residual sugars and the total acidity of the wine [130]. In another study by Heux et al. [131], a S. cerevisiae strain expressing an H₂O-NADH oxidase that reduces intracellular NADH levels was developed, significantly shifting metabolic fluxes and reducing ethanol production.
Another promising strategy is the use of global transcription machinery engineering to generate S. cerevisiae strains with reduced ethanol yield. In a recent study, this technique was applied by mutating the SPT15 gene, resulting in a strain with 34.9% reduced ethanol production while increasing CO₂ production, biomass accumulation and glycerol synthesis [132]. The reduction of the amount of glucose to be converted to ethanol in the fermentation process can be also taken as a strategy for reducing ethanol content. In the past, one way to achieve this was the introduction into S. cerevisiae of the gene that codes for a glucose oxidase that leads to the production of H2O2 and gluconic acid [133]. This approach is, however, limited, since it could lead to oxidative phenomena and alterations in the acid profile of the wine caused by the presence of these compounds.
Other potential targets for genetic engineering to produce wines with lower alcohol content include genes involved in the biosynthesis of trehalose, in central glycolysis, the oxidative pentose phosphate pathway, and the tricarboxylic acid cycle [134]. However, despite major efforts to develop engineered yeast strains, success has been limited because the use of GMOs in food production, including the wine sector, is subject to GMO restrictions in most countries and is perceived as negative by wine consumers [47].

5. Saccharomyces and Non-Saccharomyces Co-Starter Cultures

The use of co-cultures of Saccharomyces and non-Saccharomyces yeasts has become an effective strategy for producing low-alcohol wines while enhancing the sensory complexity and maintaining the desired organoleptic characteristics of the final product [135]. S. cerevisiae, the main yeast species used in winemaking, typically produces high ethanol concentrations during fermentation, but when combined with non-Saccharomyces yeasts, this can be mitigated. For example, Lachancea thermotolerans, known for its ability to produce lactic acid during fermentation, was used in co-fermentation with S. cerevisiae to reduce ethanol levels and increase wine acidity [80]. This approach resulted in lower alcohol levels and a more balanced wine, which is particularly beneficial in warmer climates where grape acidity is naturally lower [136]. S. bacillaris is a species that possesses low ethanol tolerance and produces high glycerol concentration and moderate volatile acidity production. This yeast has been studied to improve the quality of low-alcohol beverages [137]. To achieve ethanol reduction in wine, several studies proposed S. bacillaris in combination with S. cerevisiae, in mixed (co- or sequential inoculation) cultures. Laboratory and pilot scale fermentations always demonstrated a decrease of up to 1% (v/v) of ethanol and a consistent increase of glycerol [105,121]. In a study by Mestre et al. [138], the effect of co-inoculation times of selected non-Saccharomyces (H. uvarum, S. bacillaris and Candida membranaefasciens) and Saccharomyces yeasts was evaluated on the reduction of ethanol levels in wines. The timing of interactions between non-Saccharomyces and Saccharomyces yeasts during fermentation affected ethanol production, with the extent of this effect being dependent on the species co-inoculated.
Respiration of sugars by non-Saccharomyces yeasts has been recently proposed for lowering alcohol levels in wine. The development of industrial fermentation processes based on this approach requires, among other factors, the identification of yeast strains that can grow and respire in the relatively harsh conditions of grape must [139]. In the study by Sadoudi et al. [140] the effect of non-Saccharomyces strains on ethanol production when used in combination with S. cerevisiae was investigated either by simultaneous or sequential inoculation. The results showed that ethanol concentration was reduced by approximately 0.2–0.7% (v/v) compared to fermentation with S. cerevisiae alone.
In a study conducted by Contreras et al. [73], the effects of sequential inoculation of M. pulcherrima with S. cerevisiae were examined. Notably, the sequential inoculation of M. pulcherrima AWRI1149 followed by S. cerevisiae AWRI1631 gave rise to wines with a lower ethanol concentration than that achieved with S. cerevisiae (0.9 and 1.6% v/v in Chardonnay and Shiraz wines, respectively) [73]. In a subsequent study, Contreras et al. [40] found that mixed cultures of M. pulcherrima AWRI1149 and S. uvarum AWRI2846 allowed a further reduction in wine ethanol concentration compared to the same must fermented with both strains individually tested.
A further investigation conducted by Contreras et al. [141] demonstrated the potential of using non-Saccharomyces yeasts in limited aeration conditions, with the aim of producing wines with lower ethanol levels. Different species of non-Saccharomyces yeasts were tested, and the results showed that strains of T. delbrueckii and Z. bailii successfully produced wines with lower ethanol levels under limited aerobic conditions. The aeration regime significantly influenced the fermentative performance of T. delbrueckii and Z. bailii, leading to alcohol reductions of 1.5% (v/v) and 2.0% (v/v), respectively, compared to the anaerobic control with S. cerevisiae.
In a recent study conducted by Garcia et al. [101], four non-Saccharomyces yeasts, belonging to Wickerhamomyces anomalus, Meyerozyma guilliermondii, and M. pulcherrima species, were utilised in a sequential inoculation with S. cerevisiae. Results showed that the sequential fermentations produced Malvar wines with 0.8% (v/v) to 1.3% (v/v) ethanol concentrations lower than the control. In addition, these yeast combinations improved the oenological characteristics of the wines, resulting in higher levels of glycerol and a richer range of volatile higher alcohols and esters, giving a fruity and sweet profile. In a study by Jolly et al. [142] two yeast strains, S. bacillaris and W. anomalus, were tested in a small-scale winemaking trial. S. bacillaris was the most promising species for reducing alcohol content in wine production. Gobbi et al. [79] found that sequential fermentation using L. thermotolerans and S. cerevisiae (after 2 days) caused a reduction in ethanol of 0.7% (v/v).
In a study conducted by Canonico et al. [74], the initial use of immobilised non-Saccharomyces yeasts in sequential fermentation trials was evaluated in terms of reducing ethanol content in wine. Initial fermentation with S. bombicola and M. pulcherrima showed the best reductions in the final ethanol content (1.6 and 1.4% v/v, respectively). Recently, the same research group has shown that sequential fermentation of S. bombicola/S. cerevisiae under aeration conditions (20 mL/L/min for the first three days) resulted in a reduction in ethanol of 0.8% (v/v) compared to fermentation using only S. cerevisiae [25]. Looking ahead, all the above results encourage the use of co-inoculation as a valid biotechnological strategy for the production of wines with low alcohol content. Further studies should be conducted to optimise the amount of the initial inoculum of non-Saccharomyces yeasts and the timing of the sequential inoculum with S. cerevisiae for better control of the wild microbiota initially present in the must.

6. Other Methods

6.1. Arrested or Limited Fermentation

An alternative method for producing low-alcohol wines involves halting or restricting the fermentation process, through the control of factors such as fermentation temperature and duration [5,143]. When alcoholic fermentation is arrested, a considerable amount of unfermented sugars remain in the wine, making microbial stabilisation essential for the final product [144]. Therefore, the addition of sulphites or pasteurisation is necessary to extend the shelf life of the wines. Furthermore, the limited fermentation time inhibits the release of desirable aroma-active compounds, such as monoterpenes, ethyl esters, acetates, and higher alcohols, which are typically generated in large quantities by yeasts during the alcoholic fermentation in normal conditions [145].

6.2. Biomass Reduction

Biomass reduction refers to the decrease in yeast populations during must fermentation, which subsequently lowers the fermentation rate. This reduction in fermentation activity can hinder the conversion of fermentable sugars, resulting in beverages with lower ethanol content. Several studies [5,146] have investigated the production of low-alcohol cider via biomass reduction using centrifugation. The results suggest that this approach can effectively produce cider with reduced alcohol content (up to a 4% v/v reduction) and a distinctive fruity flavour profile. However, this technique frequently leads to beverages being microbiologically unstable and susceptible to spoilage due to elevated residual sugars [144].

7. Conclusions

The biotechnological approaches discussed in this review could represent concrete and valid strategies for the reduction of alcohol in wine.
However, the use of engineered yeasts (GMOs) is banned in some countries and is considered negative by consumers from a health perspective. Other techniques, such as limited fermentation and biomass reduction, lead to beverages that are microbiologically unstable and susceptible to spoilage due to the high residual sugar content. Therefore, these techniques involve the addition of sulfites or pasteurisation to extend the shelf life of wines.
Among the various approaches discussed, the industrial implementation of non-Saccharomyces or mixed cultures between non-Saccharomyces and Saccharomyces yeasts, due to its efficacy, ease of application and cost-effectiveness, could be a good biotechnological strategy for the production of low-alcohol wines. In addition, non-Saccharomyces yeasts possess specific enzymatic activities (e.g., lyase, β-glucosidase) that can contribute to the improvement of the sensory profiles of wine.
Therefore, further studies should be encouraged aimed at the selection of non-Saccharomyces yeasts, to be proposed as future starters and the optimisation of their use in the winemaking process.

Author Contributions

Conceptualisation, B.T. and M.I.; methodology, M.I. and B.T.; software, F.C. and B.T.; investigation, F.C., M.I., M.S. and B.T.; data curation, F.C., M.I., M.S. and B.T.; writing—original draft preparation, F.C., M.I., M.S. and B.T; writing—review and editing, B.T., M.I., F.C. and M.S.; visualisation, F.C.; supervision, M.I. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

No new data were created or analyzed in this study.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Rolle, L.; Englezos, V.; Torchio, F.; Cravero, F.; Río Segade, S.; Rantsiou, K.; Giacosa, S.; Gambuti, A.; Gerbi, V.; Cocolin, L. Alcohol Reduction in Red Wines by Technological and Microbiological Approaches: A Comparative Study. Aust. J. Grape Wine Res. 2018, 24, 62–74. [Google Scholar] [CrossRef]
  2. Cosme, F.; Nunes, F.M.; Filipe-Ribeiro, L. Winemaking: Advanced Technology and Flavor Research. Foods 2024, 13, 1937. [Google Scholar] [CrossRef] [PubMed]
  3. Di Renzo, M.; Letizia, F.; Di Martino, C.; Karaulli, J.; Kongoli, R.; Testa, B.; Avino, P.; Guerriero, E.; Albanese, G.; Monaco, M.; et al. Natural Fiano Wines Fermented in Stainless Steel Tanks, Oak Barrels, and Earthenware Amphora. Processes 2023, 11, 1273. [Google Scholar] [CrossRef]
  4. Pickering, G.J. Low- and Reduced-Alcohol Wine: A Review. J. Wine Res. 2000, 11, 129–144. [Google Scholar] [CrossRef]
  5. Sam, F.E.; Ma, T.-Z.; Salifu, R.; Wang, J.; Jiang, Y.-M.; Zhang, B.; Han, S.-Y. Techniques for Dealcoholization of Wines: Their Impact on Wine Phenolic Composition, Volatile Composition, and Sensory Characteristics. Foods 2021, 10, 2498. [Google Scholar] [CrossRef]
  6. Okaru, A.O.; Lachenmeier, D.W. Defining No and Low (NoLo) Alcohol Products. Nutrients 2022, 14, 3873. [Google Scholar] [CrossRef]
  7. Silva, P. Low-Alcohol and Nonalcoholic Wines: From Production to Cardiovascular Health, along with Their Economic Effects. Beverages 2024, 10, 49. [Google Scholar] [CrossRef]
  8. Corona, O.; Liguori, L.; Albanese, D.; Matteo, M.; Cinquanta, L.; Russo, P. Quality and Volatile Compounds in Red Wine at Different Degrees of Dealcoholization by Membrane Process. Eur. Food Res. Technol. 2019, 245, 2601–2611. [Google Scholar] [CrossRef]
  9. Varela, J.; Varela, C. Microbiological Strategies to Produce Beer and Wine with Reduced Ethanol Concentration. Curr. Opin. Biotechnol. 2019, 56, 88–96. [Google Scholar] [CrossRef]
  10. Longo, R.; Blackman, J.W.; Torley, P.J.; Rogiers, S.Y.; Schmidtke, L.M. Changes in Volatile Composition and Sensory Attributes of Wines during Alcohol Content Reduction. J. Sci. Food Agric. 2017, 97, 8–16. [Google Scholar] [CrossRef]
  11. Iorizzo, M.; Bagnoli, D.; Vergalito, F.; Testa, B.; Tremonte, P.; Succi, M.; Pannella, G.; Letizia, F.; Albanese, G.; Lombardi, S.J.; et al. Diversity of Fungal Communities on Cabernet and Aglianico Grapes from Vineyards Located in Southern Italy. Front. Microbiol. 2024, 15, 1399968. [Google Scholar] [CrossRef] [PubMed]
  12. Testa, B.; Coppola, F.; Iorizzo, M.; Di Renzo, M.; Coppola, R.; Succi, M. Preliminary Characterisation of Metschnikowia pulcherrima to Be Used as a Starter Culture in Red Winemaking. Beverages 2024, 10, 88. [Google Scholar] [CrossRef]
  13. Morata, A.; Escott, C.; Bañuelos, M.A.; Loira, I.; del Fresno, J.M.; González, C.; Suárez-Lepe, J.A. Contribution of Non-Saccharomyces yeasts to Wine Freshness. A Review. Biomolecules 2020, 10, 34. [Google Scholar] [CrossRef]
  14. Karaulli, J.; Xhaferaj, N.; Coppola, F.; Testa, B.; Letizia, F.; Kyçyk, O.; Kongoli, R.; Ruci, M.; Lamçe, F.; Sulaj, K.; et al. Bioprospecting of Metschnikowia pulcherrima Strains, Isolated from a Vineyard Ecosystem, as Novel Starter Cultures for Craft Beer Production. Fermentation 2024, 10, 513. [Google Scholar] [CrossRef]
  15. González-Arenzana, L.; Garijo, P.; Berlanas, C.; López-Alfaro, I.; López, R.; Santamaría, P.; Gutiérrez, A.R. Genetic and Phenotypic Intraspecific Variability of Non-Saccharomyces yeasts Populations from La Rioja Winegrowing Region (Spain). J. Appl. Microbiol. 2017, 122, 378–388. [Google Scholar] [CrossRef]
  16. Afonso, S.M.; Inês, A.; Vilela, A. Bio-Dealcoholization of Wines: Can Yeast Make Lighter Wines? Fermentation 2024, 10, 36. [Google Scholar] [CrossRef]
  17. Tilloy, V.; Cadière, A.; Ehsani, M.; Dequin, S. Reducing Alcohol Levels in Wines through Rational and Evolutionary Engineering of Saccharomyces cerevisiae. Int. J. Food Microbiol. 2015, 213, 49–58. [Google Scholar] [CrossRef]
  18. Tilloy, V.; Ortiz-Julien, A.; Dequin, S. Reduction of Ethanol Yield and Improvement of Glycerol Formation by Adaptive Evolution of the Wine Yeast Saccharomyces cerevisiae under Hyperosmotic Conditions. Appl. Environ. Microbiol. 2014, 80, 2623–2632. [Google Scholar] [CrossRef]
  19. Xu, N.; Gao, H.; Wang, Y.; Liu, C.; Hu, L.; He, A.; Jiang, W.; Xin, F. Recent Advances in Bio-Based Production of Organic Acids by Genetically Engineered Yeasts. Biochem. Eng. J. 2025, 215, 109587. [Google Scholar] [CrossRef]
  20. de Assis, L.J.; Zingali, R.B.; Masuda, C.A.; Rodrigues, S.P.; Montero-Lomelí, M. Pyruvate Decarboxylase Activity Is Regulated by the Ser/Thr Protein Phosphatase Sit4p in the Yeast Saccharomyces cerevisiae. FEMS Yeast Res. 2013, 13, 518–528. [Google Scholar] [CrossRef]
  21. Cebollero, E.; Gonzalez-Ramos, D.; Tabera, L.; Gonzalez, R. Transgenic Wine Yeast Technology Comes of Age: Is It Time for Transgenic Wine? Biotechnol. Lett. 2007, 29, 191–200. [Google Scholar] [CrossRef] [PubMed]
  22. Alonso-Del-Real, J.; Contreras-Ruiz, A.; Castiglioni, G.L.; Barrio, E.; Querol, A. The Use of Mixed Populations of Saccharomyces cerevisiae and S. Kudriavzevii to Reduce Ethanol Content in Wine: Limited Aeration, Inoculum Proportions, and Sequential Inoculation. Front. Microbiol. 2017, 8, 2087. [Google Scholar] [CrossRef]
  23. Zhao, L.-Z.; Chen, J.; Wei, X.-Y.; Lin, B.; Zheng, F.-J.; Verma, K.K.; Chen, G.-L. Response of Alcohol Fermentation Strains, Mixed Fermentation and Extremozymes Interactions on Wine Flavor. Front. Microbiol. 2025, 16, 1532539. [Google Scholar] [CrossRef]
  24. Parpinello, G.; Ricci, A.; Folegatti, B.; Patrignani, F.; Lanciotti, R.; Versari, A. Unraveling the Potential of Cryotolerant Saccharomyces eubayanus in Chardonnay White Wine Production. LWT 2020, 134, 110183. [Google Scholar] [CrossRef]
  25. Canonico, L.; Gattucci, S.; Moretti, L.; Agarbati, A.; Comitini, F.; Ciani, M. Ethanol Reduction in Montepulciano Wine: Starmerella Bombicola Sequential Fermentation at Pilot Scale Under Aeration Conditions. Foods 2025, 14, 618. [Google Scholar] [CrossRef] [PubMed]
  26. Coppola, F.; Picariello, L.; Forino, M.; Moio, L.; Gambuti, A. Comparison of Three Accelerated Oxidation Tests Applied to Red Wines with Different Chemical Composition. Molecules 2021, 26, 815. [Google Scholar] [CrossRef]
  27. Romano, P.; Braschi, G.; Siesto, G.; Patrignani, F.; Lanciotti, R. Role of Yeasts on the Sensory Component of Wines. Foods 2022, 11, 1921. [Google Scholar] [CrossRef] [PubMed]
  28. Mendoza, L.M.; Fernández de Ullivarri, M.; Raya, R. Saccharomyces cerevisiae: A Key Yeast for the Wine-Making Process. In A Closer Look at Grapes, Wines and Winemaking; Nova Science Publishers Inc.: New York, NY, USA, 2018; pp. 173–202. [Google Scholar]
  29. Chen, Y.; Jiang, J.; Song, Y.; Zang, X.; Wang, G.; Pei, Y.; Song, Y.; Qin, Y.; Liu, Y. Yeast Diversity during Spontaneous Fermentations and Oenological Characterisation of Indigenous Saccharomyces cerevisiae for Potential as Wine Starter Cultures. Microorganisms 2022, 10, 1455. [Google Scholar] [CrossRef]
  30. Testa, B.; Coppola, F.; Letizia, F.; Albanese, G.; Karaulli, J.; Ruci, M.; Pistillo, M.; Germinara, G.S.; Messia, M.C.; Succi, M.; et al. Versatility of Saccharomyces cerevisiae 41CM in the Brewery Sector: Use as a Starter for “Ale” and “Lager” Craft Beer Production. Processes 2022, 10, 2495. [Google Scholar] [CrossRef]
  31. Tronchoni, J.; Gonzalez, R.; Guindal, A.M.; Calleja, E.; Morales, P. Exploring the Suitability of Saccharomyces cerevisiae Strains for Winemaking under Aerobic Conditions. Food Microbiol. 2022, 101, 103893. [Google Scholar] [CrossRef]
  32. Kutyna, D.R.; Varela, C.; Henschke, P.A.; Chambers, P.J.; Stanley, G.A. Microbiological Approaches to Lowering Ethanol Concentration in Wine. Trends Food Sci. Technol. 2010, 21, 293–302. [Google Scholar] [CrossRef]
  33. Varela, C.; Dry, P.R.; Kutyna, D.R.; Francis, I.L.; Henschke, P.A.; Curtin, C.D.; Chambers, P.J. Strategies for Reducing Alcohol Concentration in Wine. Aust. J. Grape Wine Res. 2015, 21, 670–679. [Google Scholar] [CrossRef]
  34. Hwang, H.; Nam, S.-J. The Influence of Consumers’ Knowledge on Their Responses to Genetically Modified Foods. GM Crops Food 2021, 12, 146–157. [Google Scholar] [CrossRef]
  35. Ivit, N.N.; Longo, R.; Kemp, B. The Effect of Non-Saccharomyces and Saccharomyces Non-Cerevisiae Yeasts on Ethanol and Glycerol Levels in Wine. Fermentation 2020, 6, 77. [Google Scholar] [CrossRef]
  36. González, S.S.; Gallo, L.; Climent, M.D.; Barrio, E.; Querol, A. Enological Characterization of Natural Hybrids from Saccharomyces cerevisiae and S. kudriavzevii. Int. J. Food Microbiol. 2007, 116, 11–18. [Google Scholar] [CrossRef]
  37. Magalhães, F.; Krogerus, K.; Castillo, S.; Ortiz-Julien, A.; Dequin, S.; Gibson, B. Exploring the Potential of Saccharomyces eubayanus as a Parent for New Interspecies Hybrid Strains in Winemaking. FEMS Yeast Res. 2017, 17, fox049. [Google Scholar] [CrossRef]
  38. Álvarez, R.; Garces, F.; Louis, E.J.; Dequin, S.; Camarasa, C. Beyond S. Cerevisiae for Winemaking: Fermentation-Related Trait Diversity in the Genus Saccharomyces. Food Microbiol. 2023, 113, 104270. [Google Scholar] [CrossRef]
  39. Coloretti, F.; Zambonelli, C.; Tini, V. Characterization of Flocculent Saccharomyces Interspecific Hybrids for the Production of Sparkling Wines. Food Microbiol. 2006, 23, 672–676. [Google Scholar] [CrossRef]
  40. Contreras, A.; Curtin, C.; Varela, C. Yeast Population Dynamics Reveal a Potential “collaboration” between Metschnikowia pulcherrima and Saccharomyces uvarum for the Production of Reduced Alcohol Wines during Shiraz Fermentation. Appl. Microbiol. Biotechnol. 2015, 99, 1885–1895. [Google Scholar] [CrossRef] [PubMed]
  41. Fan, L.; Doucette, C.; McSweeney, M.; English, M.; Song, J.; Vinqvist-Tymchuk, M.; Kernaghan, G. Non-traditional Yeasts from Cool-climate Vineyards for Novel Low-alcohol Wines. Plants People Planet 2024. [Google Scholar] [CrossRef]
  42. Varela, C.; Sengler, F.; Solomon, M.; Curtin, C. Volatile Flavour Profile of Reduced Alcohol Wines Fermented with the Non-Conventional Yeast Species Metschnikowia pulcherrima and Saccharomyces uvarum. Food Chem. 2016, 209, 57–64. [Google Scholar] [CrossRef] [PubMed]
  43. Varela, C.; Barker, A.; Tran, T.; Borneman, A.; Curtin, C. Sensory Profile and Volatile Aroma Composition of Reduced Alcohol Merlot Wines Fermented with Metschnikowia pulcherrima and Saccharomyces uvarum. Int. J. Food Microbiol. 2017, 252, 1–9. [Google Scholar] [CrossRef]
  44. Orlić, S.; Sulejman, R.; Jero, A.; Herjavec, S.; Iacumin, L. Influence of Indigenous Saccharomyces paradoxus Strains on Chardonnay Wine Fermentation Aroma. Int. J. Food Sci. Technol. 2007, 42, 95–101. [Google Scholar] [CrossRef]
  45. Costantini, A.; Cravero, M.C.; Panero, L.; Bonello, F.; Vaudano, E.; Pulcini, L.; Garcia-Moruno, E. Wine Fermentation Performance of Indigenous Saccharomyces cerevisiae and Saccharomyces paradoxus Strains Isolated in a Piedmont Vineyard. Beverages 2021, 7, 30. [Google Scholar] [CrossRef]
  46. Bellon, J.R.; Schmid, F.; Capone, D.L.; Dunn, B.L.; Chambers, P.J. Introducing a New Breed of Wine Yeast: Interspecific Hybridisation between a Commercial Saccharomyces cerevisiae Wine Yeast and Saccharomyces Mikatae. PLoS ONE 2013, 8, e62053. [Google Scholar] [CrossRef]
  47. Gonzalez, R.; Guindal, A.M.; Tronchoni, J.; Morales, P. Biotechnological Approaches to Lowering the Ethanol Yield during Wine Fermentation. Biomolecules 2021, 11, 1569. [Google Scholar] [CrossRef]
  48. Coral-Medina, A.; Morrissey, J.P.; Camarasa, C. The Growth and Metabolome of Saccharomyces uvarum in Wine Fermentations Are Strongly Influenced by the Route of Nitrogen Assimilation. J. Ind. Microbiol. Biotechnol. 2023, 49, kuac025. [Google Scholar] [CrossRef]
  49. Stribny, J.; Querol, A.; Pérez-Torrado, R. Differences in Enzymatic Properties of the Saccharomyces Kudriavzevii and Saccharomyces uvarum Alcohol Acetyltransferases and Their Impact on Aroma-Active Compounds Production. Front. Microbiol. 2016, 7, 897. [Google Scholar] [CrossRef]
  50. Muratore, G.; Asmundo, C.; Lanza, C.; Caggia, C.; Licciardello, F.; Restuccia, C. Influence of Saccharomyces uvarum on Volatile Acidity, Aromatic and Sensory Profile of Malvasia Delle Lipari Wine. Food Technol. Biotechnol. 2007, 45, 101–106. [Google Scholar]
  51. Oliveira, B.M.; Barrio, E.; Querol, A.; Pérez-Torrado, R. Enhanced Enzymatic Activity of Glycerol-3-Phosphate Dehydrogenase from the Cryophilic Saccharomyces kudriavzevii. PLoS ONE 2014, 9, e87290. [Google Scholar] [CrossRef]
  52. Contreras-Ruiz, A.; Alonso-del-Real, J.; Barrio, E.; Querol, A. Saccharomyces cerevisiae Wine Strains Show a Wide Range of Competitive Abilities and Differential Nutrient Uptake Behavior in Co-Culture with S.kudriavzevii. Food Microbiol. 2023, 114, 104276. [Google Scholar] [CrossRef] [PubMed]
  53. Stribny, J.; Gamero, A.; Pérez-Torrado, R.; Querol, A. Saccharomyces Kudriavzevii and Saccharomyces uvarum Differ from Saccharomyces cerevisiae during the Production of Aroma-Active Higher Alcohols and Acetate Esters Using Their Amino Acidic Precursors. Int. J. Food Microbiol. 2015, 205, 41–46. [Google Scholar] [CrossRef] [PubMed]
  54. Libkind, D.; Hittinger, C.T.; Valério, E.; Gonçalves, C.; Dover, J.; Johnston, M.; Gonçalves, P.; Sampaio, J.P. Microbe Domestication and the Identification of the Wild Genetic Stock of Lager-Brewing Yeast. Proc. Natl. Acad. Sci. USA 2011, 108, 14539–14544. [Google Scholar] [CrossRef]
  55. Iorizzo, M.; Letizia, F.; Albanese, G.; Coppola, F.; Gambuti, A.; Testa, B.; Aversano, R.; Forino, M.; Coppola, R. Potential for Lager Beer Production from Saccharomyces cerevisiae Strains Isolated from the Vineyard Environment. Processes 2021, 9, 1628. [Google Scholar] [CrossRef]
  56. Iorizzo, M.; Coppola, F.; Letizia, F.; Testa, B.; Sorrentino, E. Role of Yeasts in the Brewing Process: Tradition and Innovation. Processes 2021, 9, 839. [Google Scholar] [CrossRef]
  57. Su, Y.; Origone, A.C.; Rodríguez, M.E.; Querol, A.; Guillamón, J.M.; Lopes, C.A. Fermentative Behaviour and Competition Capacity of Cryotolerant Saccharomyces Species in Different Nitrogen Conditions. Int. J. Food Microbiol. 2019, 291, 111–120. [Google Scholar] [CrossRef] [PubMed]
  58. Sommer, S.; Tondini, F. Sustainable Replacement Strategies for Bentonite in Wine Using Alternative Protein Fining Agents. Sustainability 2021, 13, 1860. [Google Scholar] [CrossRef]
  59. Pérez, D.; Denat, M.; Pérez-Través, L.; Heras, J.M.; Guillamón, J.M.; Ferreira, V.; Querol, A. Generation of Intra- and Interspecific Saccharomyces Hybrids with Improved Oenological and Aromatic Properties. Microb. Biotechnol. 2022, 15, 2266–2280. [Google Scholar] [CrossRef]
  60. Pérez-Torrado, R.; Barrio, E.; Querol, A. Alternative Yeasts for Winemaking: Saccharomyces Non-cerevisiae and Its Hybrids. Crit. Rev. Food Sci. Nutr. 2018, 58, 1780–1790. [Google Scholar] [CrossRef]
  61. Querol, A.; Pérez-Torrado, R.; Alonso-Del-Real, J.; Minebois, R.; Stribny, J.; Oliveira, B.M.; Barrio, E. New Trends in the Uses of Yeasts in Oenology. Adv. Food Nutr. Res. 2018, 85, 177–210. [Google Scholar] [CrossRef]
  62. Steensels, J.; Snoek, T.; Meersman, E.; Picca Nicolino, M.; Voordeckers, K.; Verstrepen, K.J. Improving Industrial Yeast Strains: Exploiting Natural and Artificial Diversity. FEMS Microbiol. Rev. 2014, 38, 947–995. [Google Scholar] [CrossRef] [PubMed]
  63. Ciani, M.; Capece, A.; Comitini, F.; Canonico, L.; Siesto, G.; Romano, P. Yeast Interactions in Inoculated Wine Fermentation. Front. Microbiol. 2016, 7, 555. [Google Scholar] [CrossRef]
  64. Renault, P.; Miot-Sertier, C.; Marullo, P.; Hernández-Orte, P.; Lagarrigue, L.; Lonvaud-Funel, A.; Bely, M. Genetic Characterization and Phenotypic Variability in Torulaspora delbrueckii Species: Potential Applications in the Wine Industry. Int. J. Food Microbiol. 2009, 134, 201–210. [Google Scholar] [CrossRef]
  65. Loira, I.; Vejarano, R.; Bañuelos, M.A.; Morata, A.; Tesfaye, W.; Uthurry, C.; Villa, A.; Cintora, I.; Suárez-Lepe, J.A. Influence of Sequential Fermentation with Torulaspora delbrueckii and Saccharomyces cerevisiae on Wine Quality. LWT-Food Sci. Technol. 2014, 59, 915–922. [Google Scholar] [CrossRef]
  66. van Breda, V.; Jolly, N.; van Wyk, J. Characterisation of Commercial and Natural Torulaspora delbrueckii Wine Yeast Strains. Int. J. Food Microbiol. 2013, 163, 80–88. [Google Scholar] [CrossRef]
  67. Ogawa, M.; Vararu, F.; Moreno-García, J.; Mauricio, J.; Moreno, J.; García-Martínez, T. Analyzing the Minor Volatilome of Torulaspora delbrueckii in an Alcoholic Fermentation. Eur. Food Res. Technol. 2022, 248, 613–624. [Google Scholar] [CrossRef]
  68. Belda, I.; Navascués, E.; Marquina, D.; Santos, A.; Calderon, F.; Benito, S. Dynamic Analysis of Physiological Properties of Torulaspora delbrueckii in Wine Fermentations and Its Incidence on Wine Quality. Appl. Microbiol. Biotechnol. 2015, 99, 1911–1922. [Google Scholar] [CrossRef]
  69. Cañas, P.M.I.; García, A.T.P.; Romero, E.G. Enhancement of Flavour Properties in Wines Using Sequential Inoculations of Non-Saccharomyces (Hansenula and Torulaspora) and Saccharomyces yeast. VITIS-J. Grapevine Res. 2011, 50, 177. [Google Scholar]
  70. Morales, P.; Rojas, V.; Quirós, M.; Gonzalez, R. The Impact of Oxygen on the Final Alcohol Content of Wine Fermented by a Mixed Starter Culture. Appl. Microbiol. Biotechnol. 2015, 99, 3993–4003. [Google Scholar] [CrossRef]
  71. Röcker, J.; Strub, S.; Ebert, K.; Grossmann, M. Usage of Different Aerobic Non-Saccharomyces yeasts and Experimental Conditions as a Tool for Reducing the Potential Ethanol Content in Wines. Eur. Food Res. Technol. 2016, 242, 2051–2070. [Google Scholar] [CrossRef]
  72. Tronchoni, J.; Curiel, J.A.; Sáenz-Navajas, M.P.; Morales, P.; de-la-Fuente-Blanco, A.; Fernández-Zurbano, P.; Ferreira, V.; Gonzalez, R. Aroma Profiling of an Aerated Fermentation of Natural Grape Must with Selected Yeast Strains at Pilot Scale. Food Microbiol. 2018, 70, 214–223. [Google Scholar] [CrossRef] [PubMed]
  73. Contreras, A.; Hidalgo, C.; Henschke, P.A.; Chambers, P.J.; Curtin, C.; Varela, C. Evaluation of Non-Saccharomyces yeasts for the Reduction of Alcohol Content in Wine. Appl. Environ. Microbiol. 2014, 80, 1670–1678. [Google Scholar] [CrossRef]
  74. Canonico, L.; Comitini, F.; Oro, L.; Ciani, M. Sequential Fermentation with Selected Immobilized Non-Saccharomyces yeast for Reduction of Ethanol Content in Wine. Front. Microbiol. 2016, 7, 278. [Google Scholar] [CrossRef]
  75. Agarbati, A.; Canonico, L.; Ciani, M.; Comitini, F. Metschnikowia pulcherrima in Cold Clarification: Biocontrol Activity and Aroma Enhancement in Verdicchio Wine. Fermentation 2023, 9, 302. [Google Scholar] [CrossRef]
  76. Muñoz-Redondo, J.M.; Puertas, B.; Cantos-Villar, E.; Jiménez-Hierro, M.J.; Carbú, M.; Garrido, C.; Ruiz-Moreno, M.J.; Moreno-Rojas, J.M. Impact of Sequential Inoculation with the Non-Saccharomyces T. Delbrueckii and M. Pulcherrima Combined with Saccharomyces cerevisiae Strains on Chemicals and Sensory Profile of Rosé Wines. J. Agric. Food Chem. 2021, 69, 1598–1609. [Google Scholar] [CrossRef]
  77. Balikci, E.K.; Tanguler, H.; Jolly, N.P.; Erten, H. Influence of Lachancea thermotolerans on Cv. Emir Wine Fermentation. Yeast 2016, 33, 313–321. [Google Scholar] [CrossRef]
  78. Fresno, J.; Morata, A.; Loira, I.; Bañuelos, M.; Escott, C.; Benito, S.; Gonzalez, C.; Suárez-Lepe, J. Use of Non-Saccharomyces in Single-Culture, Mixed and Sequential Fermentation to Improve Red Wine Quality. Eur. Food Res. Technol. 2017, 243, 2175–2185. [Google Scholar] [CrossRef]
  79. Gobbi, M.; Comitini, F.; Domizio, P.; Romani, C.; Lencioni, L.; Mannazzu, I.; Ciani, M. Lachancea thermotolerans and Saccharomyces cerevisiae in Simultaneous and Sequential Co-Fermentation: A Strategy to Enhance Acidity and Improve the Overall Quality of Wine. Food Microbiol. 2013, 33, 271–281. [Google Scholar] [CrossRef]
  80. Vicente, J.; Wang, L.; Brezina, S.; Fritsch, S.; Navascués, E.; Santos, A.; Calderón, F.; Tesfaye, W.; Marquina, D.; Rauhut, D.; et al. Enhancing Wine Fermentation through Concurrent Utilization of Lachancea thermotolerans and Lactic Acid Bacteria (Oenococcus oeni and Lactiplantibacillus plantarum) or Schizosaccharomyces pombe. Food Chem. X 2024, 24, 102054. [Google Scholar] [CrossRef]
  81. Vicente, J.; Kelanne, N.; Rodrigo-Burgos, L.; Navascués, E.; Calderón, F.; Santos, A.; Marquina, D.; Yang, B.; Benito, S. Influence of Different Lachancea thermotolerans Strains in the Wine Profile in the Era of Climate Challenge. FEMS Yeast Res. 2023, 23, foac062. [Google Scholar] [CrossRef]
  82. Hranilovic, A.; Albertin, W.; Capone, D.L.; Gallo, A.; Grbin, P.R.; Danner, L.; Bastian, S.E.P.; Masneuf-Pomarede, I.; Coulon, J.; Bely, M.; et al. Impact of Lachancea thermotolerans on Chemical Composition and Sensory Profiles of Merlot Wines. Food Chem. 2021, 349, 129015. [Google Scholar] [CrossRef] [PubMed]
  83. Blanco, P.; Rabuñal, E.; Neira, N.; Castrillo, D. Dynamic of Lachancea thermotolerans Population in Monoculture and Mixed Fermentations: Impact on Wine Characteristics. Beverages 2020, 6, 36. [Google Scholar] [CrossRef]
  84. Rossouw, D.; Bauer, F.F. Exploring the Phenotypic Space of Non-Saccharomyces Wine Yeast Biodiversity. Food Microbiol. 2016, 55, 32–46. [Google Scholar] [CrossRef] [PubMed]
  85. Mestre, M.V.; Maturano, Y.P.; Gallardo, C.; Combina, M.; Mercado, L.; Toro, M.E.; Carrau, F.; Vazquez, F.; Dellacassa, E. Impact on Sensory and Aromatic Profile of Low Ethanol Malbec Wines Fermented by Sequential Culture of Hanseniaspora uvarum and Saccharomyces cerevisiae Native Yeasts. Fermentation 2019, 5, 65. [Google Scholar] [CrossRef]
  86. Tristezza, M.; Tufariello, M.; Capozzi, V.; Spano, G.; Mita, G.; Grieco, F. The Oenological Potential of Hanseniaspora uvarum in Simultaneous and Sequential Co-Fermentation with Saccharomyces cerevisiae for Industrial Wine Production. Front. Microbiol. 2016, 7, 670. [Google Scholar] [CrossRef]
  87. Hu, K.; Jin, G.-J.; Xu, Y.-H.; Tao, Y.-S. Wine Aroma Response to Different Participation of Selected Hanseniaspora uvarum in Mixed Fermentation with Saccharomyces cerevisiae. Food Res. Int. 2018, 108, 119–127. [Google Scholar] [CrossRef] [PubMed]
  88. Hu, K.; Jin, G.-J.; Mei, W.-C.; Li, T.; Tao, Y.-S. Increase of Medium-Chain Fatty Acid Ethyl Ester Content in Mixed H. Uvarum/S. Cerevisiae Fermentation Leads to Wine Fruity Aroma Enhancement. Food Chem. 2018, 239, 495–501. [Google Scholar] [CrossRef]
  89. Maturano, Y.P.; Mestre, M.V.; Kuchen, B.; Toro, M.E.; Mercado, L.A.; Vazquez, F.; Combina, M. Optimization of Fermentation-Relevant Factors: A Strategy to Reduce Ethanol in Red Wine by Sequential Culture of Native Yeasts. Int. J. Food Microbiol. 2019, 289, 40–48. [Google Scholar] [CrossRef]
  90. Testa, B.; Coppola, F.; Lombardi, S.J.; Iorizzo, M.; Letizia, F.; Di Renzo, M.; Succi, M.; Tremonte, P. Influence of Hanseniasporauvarum AS27 on Chemical and Sensorial Characteristics of Aglianico Wine. Processes 2021, 9, 326. [Google Scholar] [CrossRef]
  91. Hu, L.; Wang, J.; Ji, X.; Liu, R.; Chen, F.; Zhang, X. Selection of Non-Saccharomyces yeasts for Orange Wine Fermentation Based on Their Enological Traits and Volatile Compounds Formation. J. Food Sci. Technol. 2018, 55, 4001–4012. [Google Scholar] [CrossRef]
  92. Filippousi, M.-E.; Chalvantzi, I.; Mallouchos, A.; Marmaras, I.; Banilas, G.; Nisiotou, A. The Use of Hanseniaspora Opuntiae to Improve ‘Sideritis’ Wine Quality, a Late-Ripening Greek Grape Variety. Foods 2024, 13, 1061. [Google Scholar] [CrossRef]
  93. Vaquero, C.; Escott, C.; Heras, J.M.; Carrau, F.; Morata, A. Co-Inoculations of Lachancea thermotolerans with Different Hanseniaspora spp.: Acidification, Aroma, Biocompatibility, and Effects of Nutrients in Wine. Food Res. Int. 2022, 161, 111891. [Google Scholar] [CrossRef] [PubMed]
  94. Testa, B.; Lombardi, S.J.; Iorizzo, M.; Letizia, F.; Martino, C.; Renzo, M.; Strollo, D.; Tremonte, P.; Pannella, G.; Ianiro, M.; et al. Use of Strain Hanseniaspora guilliermondii BF1 for Winemaking Process of White Grapes Vitis Vinifera Cv Fiano. Eur. Food Res. Technol. 2020, 246, 549–561. [Google Scholar] [CrossRef]
  95. Moreira, N.; Pina, C.; Mendes, F.; Couto, J.A.; Hogg, T.; Vasconcelos, I. Volatile Compounds Contribution of Hanseniaspora guilliermondii and Hanseniaspora uvarum during Red Wine Vinifications. Food Control 2011, 22, 662–667. [Google Scholar] [CrossRef]
  96. Lage, P.; Barbosa, C.; Mateus, B.; Vasconcelos, I.; Mendes-Faia, A.; Mendes-Ferreira, A.H. Guilliermondii Impacts Growth Kinetics and Metabolic Activity of S. Cerevisiae: The Role of Initial Nitrogen Concentration. Int. J. Food Microbiol. 2014, 172, 62–69. [Google Scholar] [CrossRef] [PubMed]
  97. Benito, S.; Hofmann, T.; Laier, M.; Lochbühler, B.; Schüttler, A.; Ebert, K.; Fritsch, S.; Röcker, J.; Rauhut, D. Effect on Quality and Composition of Riesling Wines Fermented by Sequential Inoculation with Non-Saccharomyces and Saccharomyces cerevisiae. Eur. Food Res. Technol. 2015, 241, 707–717. [Google Scholar] [CrossRef]
  98. Dutraive, O.; Benito, S.; Fritsch, S.; Beisert, B.; Patz, C.-D.; Rauhut, D. Effect of Sequential Inoculation with Non-Saccharomyces and Saccharomyces yeasts on Riesling Wine Chemical Composition. Fermentation 2019, 5, 79. [Google Scholar] [CrossRef]
  99. Vicente, J.; Calderón, F.; Santos, A.; Marquina, D.; Benito, S. High Potential of Pichia Kluyveri and Other Pichia Species in Wine Technology. Int. J. Mol. Sci. 2021, 22, 1196. [Google Scholar] [CrossRef]
  100. Barata, A.; Nobre, A.; Correia, P.; Malfeito-Ferreira, M.; Loureiro, V. Growth and 4-Ethylphenol Production by the Yeast Pichia Guilliermondii in Grape Juices. Am. J. Enol. Vitic. 2006, 57, 133–138. [Google Scholar] [CrossRef]
  101. García, M.; Esteve-Zarzoso, B.; Cabellos, J.M.; Arroyo, T. Sequential Non-Saccharomyces and Saccharomyces cerevisiae Fermentations to Reduce the Alcohol Content in Wine. Fermentation 2020, 6, 60. [Google Scholar] [CrossRef]
  102. del Mónaco, S.M.; Barda, N.B.; Rubio, N.C.; Caballero, A.C. Selection and Characterization of a Patagonian Pichia kudriavzevii for Wine Deacidification. J. Appl. Microbiol. 2014, 117, 451–464. [Google Scholar] [CrossRef]
  103. Shi, W.-K.; Wang, J.; Chen, F.-S.; Zhang, X.-Y. Effect of Issatchenkia terricola and Pichia kudriavzevii on Wine Flavor and Quality through Simultaneous and Sequential Co-Fermentation with Saccharomyces cerevisiae. LWT 2019, 116, 108477. [Google Scholar] [CrossRef]
  104. Mazzucco, M.B.; Jovanovich, M.; Rodríguez, M.E.; Oteiza, J.M.; Lopes, C.A. Pichia kudriavzevii and Saccharomyces cerevisiae Strategies for Ciders. Fermentation 2025, 11, 79. [Google Scholar] [CrossRef]
  105. Englezos, V.; Giacosa, S.; Rantsiou, K.; Rolle, L.; Cocolin, L. Starmerella bacillaris in Winemaking: Opportunities and Risks. Curr. Opin. Food Sci. 2017, 17, 30–35. [Google Scholar] [CrossRef]
  106. Englezos, V.; Rantsiou, K.; Cravero, F.; Torchio, F.; Ortiz-Julien, A.; Gerbi, V.; Rolle, L.; Cocolin, L. Starmerella bacillaris and Saccharomyces cerevisiae Mixed Fermentations to Reduce Ethanol Content in Wine. Appl. Microbiol. Biotechnol. 2016, 100, 5515–5526. [Google Scholar] [CrossRef]
  107. Englezos, V.; Di Gianvito, P.; Serafino, G.; Giacosa, S.; Cocolin, L.; Rantsiou, K. Strain Specific Starmerella bacillaris and Saccharomyces cerevisiae Interactions in Mixed Fermentations. J. Appl. Microbiol. 2024, 135, lxae085. [Google Scholar] [CrossRef] [PubMed]
  108. Moreira, L.D.P.D.; Nadai, C.; da Silva Duarte, V.; Brearley-Smith, E.J.; Marangon, M.; Vincenzi, S.; Giacomini, A.; Corich, V. Starmerella bacillaris Strains Used in Sequential Alcoholic Fermentation with Saccharomyces cerevisiae Improves Protein Stability in White Wines. Fermentation 2022, 8, 252. [Google Scholar] [CrossRef]
  109. Moreira, L.D.P.D.; Porcellato, D.; Marangon, M.; Nadai, C.; da Silva Duarte, V.; Devold, T.G.; Giacomini, A.; Corich, V. Interactions between Starmerella bacillaris and Saccharomyces cerevisiae during Sequential Fermentations Influence the Release of Yeast Mannoproteins and Impact the Protein Stability of an Unstable Wine. Food Chem. 2024, 440, 138311. [Google Scholar] [CrossRef]
  110. Suzzi, G.; Arfelli, G.; Schirone, M.; Corsetti, A.; Perpetuini, G.; Tofalo, R. Effect of Grape Indigenous Saccharomyces cerevisiae Strains on Montepulciano d’Abruzzo Red Wine Quality. Food Res. Int. 2012, 46, 22–29. [Google Scholar] [CrossRef]
  111. Whitener, M.E.B.; Stanstrup, J.; Carlin, S.; Divol, B.; Du Toit, M.; Vrhovsek, U. Effect of Non-Saccharomyces yeasts on the Volatile Chemical Profile of Shiraz Wine. Aust. J. Grape Wine Res. 2017, 23, 179–192. [Google Scholar] [CrossRef]
  112. Nadai, C.; da Silva Duarte, V.; Sica, J.; Vincenzi, S.; Carlot, M.; Giacomini, A.; Corich, V. Starmerella bacillaris Released in Vineyards at Different Concentrations Influences Wine Glycerol Content Depending on the Vinification Protocols. Foods 2023, 12, 3. [Google Scholar] [CrossRef] [PubMed]
  113. Lyu, X.; Zhou, Y.; Li, F.; Zhou, M.; Wei, C.; Lin, L.; Li, X.; Zhang, C. Improving Muscat Hamburg Wine Quality with Innovative Fermentation Strategies Using Schizosaccharomyces pombe Derived from Fermented Grains of Sauce-Flavor Baijiu. Foods 2024, 13, 1648. [Google Scholar] [CrossRef]
  114. Li, A.; Xiong, L.; Huang, Z.; Huang, R.; Tu, T.; Yu, S.; Wu, Q.; Bai, J.; Huang, Y. Enhancing the Quality and Flavor of Pomelo Wine through Sequential Fermentation with Schizosaccharomyces pombe and Saccharomyces cerevisiae: A Non-Targeted Metabolomic Analysis. Food Biosci. 2025, 63, 105585. [Google Scholar] [CrossRef]
  115. Albergaria, H.; Arneborg, N. Dominance of Saccharomyces cerevisiae in Alcoholic Fermentation Processes: Role of Physiological Fitness and Microbial Interactions. Appl. Microbiol. Biotechnol. 2016, 100, 2035–2046. [Google Scholar] [CrossRef] [PubMed]
  116. Ciani, M.; Comitini, F.; Mannazzu, I.; Domizio, P. Controlled Mixed Culture Fermentation: A New Perspective on the Use of Non-Saccharomyces yeasts in Winemaking. FEMS Yeast Res. 2010, 10, 123–133. [Google Scholar] [CrossRef]
  117. Lombardi, S.J.; Pannella, G.; Iorizzo, M.; Moreno-Arribas, M.; Tremonte, P.; Succi, M.; Sorrentino, E.; Macciola, V.; Renzo, M.; Coppola, R. Sequential Inoculum of Hanseniaspora guilliermondii and Saccharomyces cerevisiae for Winemaking Campanino on an Industrial Scale. World J. Microbiol. Biotechnol. 2018, 34, 161. [Google Scholar] [CrossRef]
  118. Padilla, B.; Gil, J.V.; Manzanares, P. Past and Future of Non-Saccharomyces yeasts: From Spoilage Microorganisms to Biotechnological Tools for Improving Wine Aroma Complexity. Front. Microbiol. 2016, 7, 411. [Google Scholar] [CrossRef]
  119. Alonso, A.; Belda, I.; Santos, A.; Navascués, E.; Marquina, D. Advances in the Control of the Spoilage Caused by Zygosaccharomyces Species on Sweet Wines and Concentrated Grape Musts. Food Control 2015, 51, 129–134. [Google Scholar] [CrossRef]
  120. Maicas, S.; Mateo, J.J. The Life of Saccharomyces and Non-Saccharomyces yeasts in Drinking Wine. Microorganisms 2023, 11, 1178. [Google Scholar] [CrossRef]
  121. Rantsiou, K.; Englezos, V.; Torchio, F.; Risse, P.-A.; Francesco, C.; Gerbi, V.; Rolle, L.; Cocolin, L. Modeling of the Fermentation Behavior of Starmerella bacillaris. Am. J. Enol. Vitic. 2017, 68, 378–385. [Google Scholar] [CrossRef]
  122. Belda, I.; Navascués, E.; Marquina, D.; Santos, A.; Calderón, F.; Benito, S. Outlining the Influence of Non-Conventional Yeasts in Wine Ageing over Lees. Yeast 2016, 33, 329–338. [Google Scholar] [CrossRef] [PubMed]
  123. Mestre Furlani, M.V.; Maturano, Y.P.; Combina, M.; Mercado, L.A.; Toro, M.E.; Vazquez, F. Selection of Non-Saccharomyces yeasts to Be Used in Grape Musts with High Alcoholic Potential: A Strategy to Obtain Wines with Reduced Ethanol Content. FEMS Yeast Res. 2017, 17, fox010. [Google Scholar] [CrossRef]
  124. Escott, C.; Del Fresno, J.M.; Loira, I.; Morata, A.; Suárez-Lepe, J.A. Zygosaccharomyces Rouxii: Control Strategies and Applications in Food and Winemaking. Fermentation 2018, 4, 69. [Google Scholar] [CrossRef]
  125. Zhu, X.; Navarro, Y.; Mas, A.; Torija, M.-J.; Beltran, G. A Rapid Method for Selecting Non-Saccharomyces Strains with a Low Ethanol Yield. Microorganisms 2020, 8, 658. [Google Scholar] [CrossRef] [PubMed]
  126. Jolly, N.P.; Varela, C.; Pretorius, I.S. Not Your Ordinary Yeast: Non-Saccharomyces yeasts in Wine Production Uncovered. FEMS Yeast Res. 2014, 14, 215–237. [Google Scholar] [CrossRef]
  127. Molina-Espeja, P. Next Generation Winemakers: Genetic Engineering in Saccharomyces cerevisiae for Trendy Challenges. Bioengineering 2020, 7, 128. [Google Scholar] [CrossRef]
  128. Kessi-Pérez, E.I.; Gómez, M.; Farías, W.; García, V.; Ganga, M.A.; Querol, A.; Martínez, C. Genetically Improved Yeast Strains with Lower Ethanol Yield for the Wine Industry Generated Through a Two-Round Breeding Program. J. Fungi 2025, 11, 137. [Google Scholar] [CrossRef]
  129. Goold, H.D.; Kroukamp, H.; Williams, T.C.; Paulsen, I.T.; Varela, C.; Pretorius, I.S. Yeast’s Balancing Act between Ethanol and Glycerol Production in Low-Alcohol Wines. Microb. Biotechnol. 2017, 10, 264–278. [Google Scholar] [CrossRef]
  130. Cuello, R.A.; Flores Montero, K.J.; Mercado, L.A.; Combina, M.; Ciklic, I.F. Construction of Low-Ethanol-Wine Yeasts through Partial Deletion of the Saccharomyces cerevisiae PDC2 Gene. AMB Express 2017, 7, 67. [Google Scholar] [CrossRef]
  131. Heux, S.; Cachon, R.; Dequin, S. Cofactor Engineering in Saccharomyces cerevisiae: Expression of a H2O-Forming NADH Oxidase and Impact on Redox Metabolism. Metab. Eng. 2006, 8, 303–314. [Google Scholar] [CrossRef]
  132. Du, Q.; Liu, Y.; Song, Y.; Qin, Y. Creation of a Low-Alcohol-Production Yeast by a Mutated SPT15 Transcription Regulator Triggers Transcriptional and Metabolic Changes During Wine Fermentation. Front. Microbiol. 2020, 11, 597828. [Google Scholar] [CrossRef] [PubMed]
  133. Malherbe, D.F.; du Toit, M.; Cordero Otero, R.R.; van Rensburg, P.; Pretorius, I.S. Expression of the Aspergillus Niger Glucose Oxidase Gene in Saccharomyces cerevisiae and Its Potential Applications in Wine Production. Appl. Microbiol. Biotechnol. 2003, 61, 502–511. [Google Scholar] [CrossRef] [PubMed]
  134. Rossouw, D.; Heyns, E.H.; Setati, M.E.; Bosch, S.; Bauer, F.F. Adjustment of Trehalose Metabolism in Wine Saccharomyces cerevisiae Strains to Modify Ethanol Yields. Appl. Environ. Microbiol. 2013, 79, 5197–5207. [Google Scholar] [CrossRef]
  135. Xu, A.; Xiao, Y.; He, Z.; Liu, J.; Wang, Y.; Gao, B.; Chang, J.; Zhu, D. Use of Non-Saccharomyces yeast Co-Fermentation with Saccharomyces cerevisiae to Improve the Polyphenol and Volatile Aroma Compound Contents in Nanfeng Tangerine Wines. J. Fungi 2022, 8, 128. [Google Scholar] [CrossRef]
  136. Peskova, I.; Tanashchuk, T.; Ostroukhova, E.; Slastya, E.; Levchenko, S.; Lutkova, N. Prospects of Using Lachancea thermotolerans Yeast in Winemaking. E3S Web Conf. 2021, 247, 01012. [Google Scholar] [CrossRef]
  137. Lemos Junior, W.J.F.; da Silva Duarte, V.; Treu, L.; Campanaro, S.; Nadai, C.; Giacomini, A.; Corich, V. Whole Genome Comparison of Two Starmerella bacillaris Strains with Other Wine Yeasts Uncovers Genes Involved in Modulating Important Winemaking Traits. FEMS Yeast Res. 2018, 18, foy069. [Google Scholar] [CrossRef]
  138. Mestre, V.; Maturano, P.; Mercado, L.; Toro, M.; Vazquez, F.; Combina, M. Evaluation of Different Co-Inoculation Time of Non-Saccharomyces/Saccharomyces yeasts in Order to Obtain Reduced Ethanol Wines. BIO Web Conf. 2016, 7, 02025. [Google Scholar] [CrossRef]
  139. Quirós, M.; Rojas, V.; Gonzalez, R.; Morales, P. Selection of Non-Saccharomyces yeast Strains for Reducing Alcohol Levels in Wine by Sugar Respiration. Int. J. Food Microbiol. 2014, 181, 85–91. [Google Scholar] [CrossRef]
  140. Sadoudi, M.; Tourdot-Maréchal, R.; Rousseaux, S.; Steyer, D.; Gallardo-Chacón, J.-J.; Ballester, J.; Vichi, S.; Guérin-Schneider, R.; Caixach, J.; Alexandre, H. Yeast-Yeast Interactions Revealed by Aromatic Profile Analysis of Sauvignon Blanc Wine Fermented by Single or Co-Culture of Non-Saccharomyces and Saccharomyces yeasts. Food Microbiol. 2012, 32, 243–253. [Google Scholar] [CrossRef]
  141. Contreras, A.; Hidalgo, C.; Schmidt, S.; Henschke, P.A.; Curtin, C.; Varela, C. The Application of Non-Saccharomyces yeast in Fermentations with Limited Aeration as a Strategy for the Production of Wine with Reduced Alcohol Content. Int. J. Food Microbiol. 2015, 205, 7–15. [Google Scholar] [CrossRef]
  142. Jolly, N.; Mehlomakulu, N.N.; Nortje, S.; Beukes, L.; Hoff, J.; Booyse, M.; Erten, H. Non-Saccharomyces yeast for Lowering Wine Alcohol Levels: Partial Aeration versus Standard Conditions. FEMS Yeast Res. 2022, 22, foac002. [Google Scholar] [CrossRef] [PubMed]
  143. Strejc, J.; Kyselová, L.; Karabin, M.; Silva, J.; Brányik, T. Production of Alcohol-free Beer with Elevated Amounts of Flavouring Compounds Using Lager Yeast Mutants. J. Inst. Brew. 2013, 119, 149–155. [Google Scholar] [CrossRef]
  144. Ferreira, M. Yeasts and Wine Off-Flavours: A Technological Perspective. Ann. Microbiol. 2011, 61, 95–102. [Google Scholar] [CrossRef]
  145. Capece, A.; Romano, P. Yeasts and Their Metabolic Impact on Wine Flavour; Springer: New York, NY, USA, 2019; pp. 43–80. ISBN 978-1-4939-9780-0. [Google Scholar]
  146. Nogueira, A.; Quéré, J.M.L.; Gestin, P.; Michel, A.; Wosiacki, G.; Drilleau, J.F. Slow Fermentation in French Cider Processing Due to Partial Biomass Reduction. J. Inst. Brew. 2008, 114, 102–110. [Google Scholar] [CrossRef]
Fermentation 11 00159 g001
Figure 1. Biotechnology approaches for low-alcohol wine production.
Figure 1. Biotechnology approaches for low-alcohol wine production.
Fermentation 11 00159 g001
Table 1. Ethanol reduction in wines produced with Saccharomyces non-cerevisiae yeasts.
Table 1. Ethanol reduction in wines produced with Saccharomyces non-cerevisiae yeasts.
Saccharomyces spp.Ethanol ReductionGrape VarietyCommentsReferences
Saccharomyces kudriavzevii1.9–3%Tempranillo, Macabeo, Synthetic grape must↑ higher alcohols (2-phenylethanol, isobutanol)[22,36]
Saccharomyces eubayanus0.4%Sauvignon blanc, Macabeu; Synthetic grape must↑ higher alcohols (2-phenylethanol), ↑ glycerol[37,38]
Saccharomyces uvarum0.5–1.7%Trebbiano, Chardonnay, Pinot, Shiraz, Merlot↑ higher alcohols (2-phenylethanol); ↑ esters (2-phenylethyl acetate, ethyl 2-methyl butanoate); ↑ glycerol; ↑ volatile acid (2-Methyl butanoic acid)[39,40,41,42,43]
Saccharomyces paradoxus0.3–0.56%Chardonnay, Grignolino↑ higher alcohols (1-propanol, hexanol, 2-phenylethanol, cis-3-hexenol)[44,45]
Saccharomyces mikatae0.5%Chardonnay; Synthetic grape must↑ higher alcohols (2-phenyl ethyl alcohol, 4-hydroxybenzene ethanol); ↑ glycerol[38,46]
↑ increase of concentration.
Table 2. Ethanol reduction in wines produced with non-Saccharomyces yeasts.
Table 2. Ethanol reduction in wines produced with non-Saccharomyces yeasts.
Non-Saccharomyces spp.Ethanol ReductionGrape VarietyCommentsReferences
Torulaspora delbrueckii0.3–1.3%Tempranillo; Chenin blanc–Chardonnay blend; Airen, Synthetic grape must↑ higher alcohols (isoamyl alcohol, 2-phenylethanol, isobutanol); ↑ ester (phenylethyl acetate, ethyl lactate, 2-phenylethyl acetate, isoamyl acetate, ethyl heptanoate); ↑ glycerol; ↑ organic acid (succinic acid); ↑ aldheydes (nonanal, decanal)[64,65,66,67,68,69]
Metschnikowia pulcherrima0.8–7.5%Chardonnay, Shiraz, Verdicchio, Viura–Malvasìa blend, Riesling, Merlot, Synthetic grape must, Aglianico, Verdicchio, Garnacha tinta–Cabernet Sauvignon blend↑ higher alcohols (methyl propanol, 2-methyl butanol, 3-methyl butanol, isobutanol, 3-methylbutanol); ↑ esters (2-methylbutyl acetate, ethyl 2-methyl propanoate, ethyl acetate, ethyl propionate, ethyl octanoate, ethyl hexanoate); ↑ glycerol; ↑ terpene (geraniol)[12,43,70,71,72,73,74,75,76]
Lachancea thermotolerans0.5–2.6%Emir, Tempranillo, Sangiovese–Cabernet Sauvignon blend; Merlot, Treixadura, Mencía ↑ higher alcohols (butanol, isobutanol, 2-methyl butanol, propanol, hexanol); ↑ glycerol; increased of esters (ethyl lactate, ethyl acetate, ethyl 3-hydroxybutyrate, ethyl octanoate, ethyl hexanoate); ↑ terpene (linalool)[77,78,79,80,81,82,83]
Hanseniaspora uvarum0.4–6.7%Malbec, Pinotage, Negroamaro, Ecolly, Cabernet Sauvignon, Aglianico↑ glycerol; ↑ higher alcohols (2-methyl-1-propanol, 1-pentanol, 1-eexanol, 3-ethoxy-1-propanol, benzyl alcohol, 2-pentanol); ↑ esters (ethyl hexanoate, ethyl octanoate, ethyl decanoate, ethyl lactate, isoamyl acetate; ethyl acetate); ↑ terpenes (linalool, nerol, nonanol)[84,85,86,87,88,89,90]
Hanseniaspora opuntiae0.6–1.25%Negroamaro, Pinotage, Sauvignon Blanc, Sideritis, Airén↑ higher alcohols (1-pentanol, 1-hexanol, 2-methyl-1-propanol, 2-nonanol); ↑ esters (phenethyl acetate, isoamyl acetate, ethyl hexanoate, ethyl caprylate, ethyl acetate, methyl acetate); ↑ organic acid (lactic acid)[84,91,92,93]
Hanseniaspora guilliermondii3.3–3.3%Fiano, Campanino↑ higher alcohols (1-propanol, 2-phenylethanol, 2–3-butanediol, 2-methyl-butanol); ↑ esters (2-phenylethyl acetate); ↑ organic acid (3-methylthio propionic acid); ↑ terpenes (nerol, α-terpineol)[90,94,95,96]
Pichia kluyveri0.16–5%Riesling↑ esters (ethyl hexanoate, ethyl octanoate); ↓ isovaleric acid; ↑ esters (ethyl butanoate, ethyl octanoate); ↑ terpenes (linalool oxide, hotrienol)[71,97,98,99]
Meyerozyma guilliermondii0.8–2%Riesling, Synthetic grape must, White Malvar ↓ isovaleric acid; ↑ phenol (4-ethyl-phenol) ↑ higher alcohols (1-butanol, isoamyl alcohol, β-phenylethyl alcohol, isobutanol); ↑ esters (isoamyl acetate); ↑ ketone (acetoin)[71,99,100,101]
Pichia kudriavzevii0.16–2.5%Cabernet Sauvignon, Pinot noir, Cider↑ higher alcohols (n-pentanol, 1-phenylethanol, isoamyl alcohol, 1 octanol); ↑ glycerol; ↑ esters (ethyl propanoate, ethyl octanoate, ethyl acetate, benzyl acetate); ↑ terpenes (limonene, linalool)[102,103,104]
Starmerella bacillaris0.5–4%Barbera, Riesling, Montepulciano, Nebbiolo, Pinot grigio; Sauvignon blanc, Manzoni bianco, Raboso Piave↑ glycerol; ↑ higher alcohols (isoamyl alcohols, 1-octanol, phenylethyl alcohol, 2-phenylethanol); ↑ esters (ethyl hexanoate, ethyl acetate, ethyl octanoate); ↑ terpenes (linalool, geraniol) [1,71,105,106,107,108,109,110,111,112]
Starmerella bombicola0.8–2.24%Verdicchio, Synthetic grape must, Montepulciano↑ esters (ethyl acetate, isoamyl acetate); ↑ glycerol; ↑ terpenes (linalool); ↑ higher alcohols (β-phenyl ethanol, isobutanol)[25,33,70,109,110,111]
Schizosaccharomyces pombe0.2–1%Airen, Tempranillo, Zhenlong pomelo↑ higher alcohols (1-propanol, isobutanol, isoamyl alcohol, phenylethanol); ↓ organic acid (malic acid)[80,97,113,114]
↑ increase of concentration; ↓ decrease of concentration.
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Testa, B.; Coppola, F.; Succi, M.; Iorizzo, M. Biotechnological Strategies for Ethanol Reduction in Wine. Fermentation 2025, 11, 159. https://doi.org/10.3390/fermentation11030159

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Testa B, Coppola F, Succi M, Iorizzo M. Biotechnological Strategies for Ethanol Reduction in Wine. Fermentation. 2025; 11(3):159. https://doi.org/10.3390/fermentation11030159

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Testa, Bruno, Francesca Coppola, Mariantonietta Succi, and Massimo Iorizzo. 2025. "Biotechnological Strategies for Ethanol Reduction in Wine" Fermentation 11, no. 3: 159. https://doi.org/10.3390/fermentation11030159

APA Style

Testa, B., Coppola, F., Succi, M., & Iorizzo, M. (2025). Biotechnological Strategies for Ethanol Reduction in Wine. Fermentation, 11(3), 159. https://doi.org/10.3390/fermentation11030159

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