Optimal trade-off between boosted tolerance and growth fitness during adaptive evolution of yeast to ethanol shocks.

BACKGROUND: The selection of Saccharomyces cerevisiae strains with higher alcohol tolerance can potentially increase the industrial production of ethanol fuel. However, the design of selection protocols to obtain bioethanol yeasts with higher alcohol tolerance poses the challenge of improving industrial strains that are already robust to high ethanol levels. Furthermore, yeasts subjected to mutagenesis and selection, or laboratory evolution, often present adaptation trade-offs wherein higher stress tolerance is attained at the expense of growth and fermentation performance. Although these undesirable side effects are often associated with acute selection regimes, the utility of using harsh ethanol treatments to obtain robust ethanologenic yeasts still has not been fully investigated. RESULTS: We conducted an adaptive laboratory evolution by challenging four populations (P1-P4) of the Brazilian bioethanol yeast, Saccharomyces cerevisiae PE-2_H4, through 68-82 cycles of 2-h ethanol shocks (19-30% v/v) and outgrowths. Colonies isolated from the final evolved populations (P1c-P4c) were subjected to whole-genome sequencing, revealing mutations in genes enriched for the cAMP/PKA and trehalose degradation pathways. Fitness analyses of the isolated clones P1c-P3c and reverse-engineered strains demonstrated that mutations were primarily selected for cell viability under ethanol stress, at the cost of decreased growth rates in cultures with or without ethanol. Under this selection regime for stress survival, the population P4 evolved a protective snowflake phenotype resulting from BUD3 disruption. Despite marked adaptation trade-offs, the combination of reverse-engineered mutations cyr1A1474T/usv1Δ conferred 5.46% higher fitness than the parental PE-2_H4 for propagation in 8% (v/v) ethanol, with only a 1.07% fitness cost in a culture medium without alcohol. The cyr1A1474T/usv1Δ strain and evolved P1c displayed robust fermentations of sugarcane molasses using cell recycling and sulfuric acid treatments, mimicking Brazilian bioethanol production. CONCLUSIONS: Our study combined genomic, mutational, and fitness analyses to understand the genetic underpinnings of yeast evolution to ethanol shocks. Although fitness analyses revealed that most evolved mutations impose a cost for cell propagation, combination of key mutations cyr1A1474T/usv1Δ endowed yeasts with higher tolerance for growth in the presence of ethanol. Moreover, alleles selected for acute stress survival comprising the P1c genotype conferred stress tolerance and optimal performance under conditions simulating the Brazilian industrial ethanol production.

Fermentation Technology and Functional Foods.

Food is an integral part of our civilization. It is a cultural phenomenon that, while having evolved, is associated with societal traditions and identity. This work analyzes studies conducted to highlight the health properties of the most common ethnic foods. Although these foods were originally created from the need to preserve perishable produce, presently, we know that the fermentation process makes them nutritionally more complete. The basis of these transformations lies in that vast range of prokaryotic and eukaryotic microorganisms that, similar to small biochemical factories, can transform the initial nutrients into metabolically more active biomolecules through fermentation. Although naturally occurring microbes work together for mutual benefit, environmental conditions enhance or inhibit their development. Starting from a selection of microorganisms naturally present on a substrate, we attempt to select the most suitable species to obtain a fermented food with the best nutritional qualities and the richest in nutraceuticals.

Sake yeast symbiosis with lactic acid bacteria and alcoholic fermentation.

The yeast Saccharomyces cerevisiae plays a pivotal role in the production of fermented foods by converting sugars in ingredients into ethanol through alcoholic fermentation. However, how accurate is our understanding of its biological significance? Although yeast is essential to produce alcoholic beverages and bioethanol, yeast does not yield ethanol for humankind. Yeast obtains energy in the form of ATP for its own vital processes through alcoholic fermentation, which generates ethanol as a byproduct. The production of ethanol may have more significance for yeast, since many other organisms do not produce ethanol, a highly toxic substance, to obtain energy. The key to address this issue has not been found using conventional microbiology, where yeasts are isolated and cultured in pure form. This review focuses on a possible novel role of yeast alcohol fermentation, which is revealed through our recent studies of microbial interactions.

Single versus dual inoculation with indigenous Saccharomyces cerevisiae strains in winemaking.

Indigenous Saccharomyces cerevisiae strains and their combinations may be used to diversify wines and add complexity to sensory profiles. Here, two S. cerevisiae strains that represent regional genetic and phenotypic specificities for two major winegrowing areas of Greece were used in single- and mixed-culture fermentations. The kinetics and metabolic activities of the strains were analyzed to evaluate the influence of each strain individually or in combination on wine quality. The two strains differentially affected the kinetics and the outcome of fermentation. They showed significant differences in the production of important metabolites that strongly affect the organoleptic profile of wines, such as volatile acidity, acetaldehyde, certain esters, and terpenes. Furthermore, the chemical and sensory profiles of wines produced by single cultures were different from those fermented by mixed-culture inoculum. The concentration of certain metabolites was enhanced (e.g. isoamyl acetate, 1-heptanol), while others were suppressed (e.g. hexyl acetate, octyl acetate). Results highlight the potential worth of indigenous S. cerevisiae strains to differentiate local wines. The mixed-culture S. cerevisiae inoculum was shown to generate novel wine characteristics, as compared to single cultures, thus offering alternatives to further diversify wines and increase their complexity.

Influence of spontaneous, "pied de cuve" and commercial dry yeast fermentation strategies on wine molecular composition and sensory properties.

While most producers in recent decades have relied on commercial yeasts (ADY) as their primary choice given their reliability and reproducibility, the fear of standardising the taste and properties of wine has led to the employment of alternative strategies that involve autochthonous yeasts such as pied de cuve (PdC) and spontaneous fermentation (SF). However, the impact of different fermentation strategies on wine has been a subject of debate and speculation. Consequently, this study describes, for the first time, the differences between the three kinds of fermentation at the metabolomic, chemical, and sensory levels in two wines: Chardonnay and Pinot Noir. The results showed how the yeast chosen significantly impacted the molecular composition of the wines, as revealed by metabolomic analysis that identified biomarkers with varying chemical compositions according to the fermentation modality. Notably, higher numbers of lipid markers were found for SF and PdC than ADY, which contained more peptides. Key molecules from the metabolic amino acid pathway, which are addressed in this article, showed evidence of such variations. In addition, the analysis of volatile aromatic compounds revealed an increase in groups of compounds specific to each fermentation. The sensorial analysis of Chardonnay wine showed a more qualitative sensory outcome (Higher fruit intensity) for ADY and SF compared to PdC. Our finding challenges the common speculation among wine producers that autochthonous yeast fermentations may offer greater complexity and uniqueness in comparison to commercial yeast fermentations.

Unlocking the secrets of peptide transport in wine yeast: insights into oligopeptide transporter functions and nitrogen source preferences.

IMPORTANCE: Limited nitrogen supply can prevent the completion of alcoholic fermentation. Supplementation through peptides as an alternative, natural source of nitrogen for yeast offers an interesting solution for this issue. In this work, the S. cerevisiae peptide transporters of the Opt and Fot families were studied. We demonstrated that Fot and Opt2 have a broader peptide length preference than previously reported, enabling yeasts to acquire sufficient nitrogen from peptides without requiring additional ammonia or amino acids to complete fermentation. On the contrary, Opt1 was unable to consume any peptide in the given conditions, whereas it has been described elsewhere as the main peptide transporter for peptides longer than three amino acid residues in experiments in laboratory conditions. This controversy signifies the need in applied sciences for approaching experimental conditions to those prevalent in the industry for its more accurate characterization. Altogether, this work provides further evidence of the importance of peptides as a nitrogen source for yeast and their consequent positive impact on fermentation kinetics.

Volatile Compound Release from Oak Chips in Model Wine Media: Combined Influence of Toasting Degree, Size, Time of Contact, and Ethanol Content.

The effects of size, toasting degree, and time of contact on the release of volatile compounds from Quercus alba (L.) chips during a simulated fermentation and post-fermentative process were studied. The results obtained indicated that the large-size chips favored the release of furfural and furfuryl alcohol, while the small ones increased the concentration of cyclotene and maltol. The interaction between chip size and time of contact showed that the small-size chips are more sensitive to the increase of ethanol concentration for the extraction rate of some compounds (furfural, vanillin, maltol, cyclotene, whiskey lactones, and eugenol) compared to the large-size ones, increasing their concentrations at the end of maceration. The toasting degree of oak chips had a different influence on the volatile compounds studied. Cyclotene and guaiacol concentrations increased with the toasting intensity, whereas the extracted concentration of all compounds increased from light to medium-toasted chips, except for eugenol, and then decreased by further increasing the toasting level for 5-methylfurfural, whiskey lactones, eugenol, and only using high-level toasted chips for furfuryl alcohol, maltol, and vanillin. A possible protection effect of the chip size toward the possible degradation or volatilization losses of furfural for high toasting degrees was observed.

Evolution of Markers Involved in the Fresh Mushroom Off-Flavor in Wine During Alcoholic Fermentation.

The fresh mushroom off-flavor (FMOff) is due to several C8 compounds such as 1-octen-3-one, 1-octen-3-ol and 1-hydroxyoctan-3-one, among others. Recently, glycosidic precursors of some FMOff compounds have been identified in grape musts, but the evolution of such compounds during alcoholic fermentation (AF) remains poorly studied. Therefore, the aim of this work was to monitor both FMOff glycosidic precursors and volatile compounds during AF by comparing healthy and Crustomyces subabruptus-contaminated musts. For the first time, glycosidic analysis revealed the presence of 1-hydroxyoctan-3-one glycosides in the laboratory-contaminated musts, together with other FMOff glycosidic fractions already described in the literature. During AF, the FMOff glycosidic fraction decreased, even more in the case of 1-hydroxyoctan-3-one precursors. For the volatile FMOff compounds, their evolutions were both compound- and matrix-dependent except for 1-hydroxyoctan-3-one, which seemed to reach an identical threshold concentration in wine regardless of its initial level in contaminated musts.

To each their own: Delving into the vitaminic preferences of non-Saccharomyces wine yeasts.

Considering the growing interest in non-Saccharomyces wine yeasts, and notably in the context of mixed fermentations with S. cerevisiae, understanding their nutritional behaviors is essential to ensure better management of these fermentations. The vitaminic consumption of three non-Saccharomyces yeasts (Starmerella bacillaris, Metschnikowia pulcherrima and Torulaspora delbrueckii) was investigated during their growth in wine-like conditions, providing initial evidence that they consume different vitamers. The vitamin consumption profiles during their growth highlighted releases of certain vitamers by the yeasts before re-assimilation, strongly suggesting the existence of synthesis pathways. Not only did the essential character of vitamin B1, in particular, appear to be a trait common to these yeasts, since all its vitamers are consumed, this investigation also provided evidence of the existence of species-dependent preferences for their vitaminic sources. These different behaviors were quite striking in certain vitamers, as was observed in nicotinamide: while it was consumed by T. delbrueckii, it was left untouched by S. bacillaris and produced by M. pulcherrima during growth. Furthermore, this offers grounds for further investigation into these yeasts' requirements, and provides the first tool for managing vitamin resources during mixed fermentations with S. cerevisiae, and for preventing nutritive deficiencies from occurring.

Study of Different Types of Fermentation in Wine-Making Process and Considering Aromatic Substances and Organic Acid.

Fermentation is being utilized since ages as a safe process for food preservation. Fermentation in winemaking is the process which converts grapes juice into wine. Alcoholic fermentation is the principle for the manufacturing of alcoholic beverages like wine and beer. Several by-products are produced by the alcoholic fermentation of organic carbon sources by Saccharomyces cerevisiae. The secondary bacterial fermentation is malolactic fermentation which is carried out in most red and some kinds of white wines. Malolactic fermentation causes acid reduction, flavor modification, and also is accountable for microbial stability. The present searching was carried out by the keywords in major indexing systems including PubMed/MEDLINE, Scopus, Institute for Scientific Web of Science and the search engine of Google Scholar. The keywords were alcoholic fermentation, malolactic fermentation, bottle fermentation, carbonic maceration, Saccharomyces, Lactobacillus, Oenococcus oeni, aromatic substances, flavour quality, terpenes, esters, lactones, aldehydes, organic acid, pyrazines, mercaptans, and traditional fermented food. This review was done to highlight the significance, and introduce different kinds of fermentations carried out in wine-making process, specially alcoholic and malolactic fermentation, survey on aromatic substances, flavour quality, and organic acids, and introduce the most notable traditional fermented food in China.

How Do Different Ingredients and Additives Affect the Production Steps and the Bioactive Potential of Mead?

Mead is a fermented alcoholic beverage that is made from honey diluted in water and commonly with the addition of other ingredients. The chemical characteristics of mead are closely related to the ingredients and additives that are used in its preparation, especially the type of honey, yeast strain and prefermentation nutrients, as well as herbs, spices and/or fruits. These additives can affect not only the fermentation process, in particular the yeast activity, the formation of metabolites and fermentation time, but also the bioactive potential of the mead, which mainly depends on phenolic compounds. Scientific studies have shown that the mead with added different plant species contains considerable amounts of different classes of polyphenols, which have important biological activities. Within this context, this review study seeks to investigate how different ingredients and additives can affect each of the stages of the preparation of mead, as well as its bioactive potential, in order to understand the effects on its chemical composition, and thus add greater commercial value to this beverage.

Management of in-Amphora "Trebbiano Toscano" Wine Production: Selection of Indigenous Saccharomyces cerevisiae Strains and Influence on the Phenolic and Sensory Profile.

The use of earthenware amphorae in winemaking can give wines unique attributes enhancing their typicity. Therefore, in this study, spontaneous and inoculated in-amphora fermentations of Trebbiano Toscano grape must were monitored to assess the Saccharomyces cerevisiae strains occurring in each fermentation as well as the chemical characteristics of the wines. Strain typing via Interdelta analyses pointed out that the commercial starters did not dominate, showing 24% and 13% implantation percentages, and that 20 indigenous strains were present at different percentages, ranging from 2 to 20%, in inoculated and spontaneous fermentations. The assessment of the technical characteristics of the indigenous strains via fermentations at lab and pilot scale (20 L amphorae) and the sensory analysis of the experimental wines allowed for the selection of two indigenous strains to be used as starter cultures in comparison to a commercial strain in 300-L-amphorae vinifications in the cellar. The observed fermentative performances and sensory analysis of the experimental wines highlighted that one indigenous S. cerevisiae strain dominated the process and conferred distinctive sensory characteristics to the Trebbiano Toscano wine, demonstrating its effectiveness in managing the in-amphora fermentations. In addition, the results demonstrated the ability of amphorae to protect the polyphenolic compounds from oxidation during wine ageing. Indeed, the concentration of both hydroxycinnamic acids and flavonols decreased, with an average reduction of 30% and 14%, respectively, while hydroxybenzoic acids remained unchanged.

Enological Suitability of Indigenous Yeast Strains for 'Verdejo' Wine Production.

The use of indigenous yeasts for the production of wines is a tool to defend the typicity of a particular region. The selection of appropriate indigenous yeasts ensures the maintenance of oenological characteristics by simulating spontaneous alcoholic fermentation (AF) while avoiding the risks of stuck or sluggish fermentations. In this study, autochthonous yeasts from Verdejo grape juice (Appellation of Origin Rueda) were selected, identified, and characterized to exploit the characteristics of the 'terroir'. The fermentation capacity of seven strains was studied individually at the laboratory scale. The most suitable strains (Saccharomyces cerevisiae: Sacch 1, Sacch 2, Sacch 4, and Sacch 6) and Sacch 6 co-inoculated with Metschnikowia pulcherrima were characterized at the pilot scale. The fermentation kinetics, bioproduct release, volatile composition, and sensory profile of the wines were evaluated. Significant differences were found, especially in the aroma profile. In particular, Sacch 6 and Sacch 6 co-inoculated with M. pulcherrima produced higher amounts of ethyl esters and acetates and lower amounts of higher alcohols than the spontaneous AF. Wines inoculated with indigenous yeasts had higher sensory scores for fruit aromas and overall rating. The selection of indigenous yeasts improved the aroma of Verdejo wines and could contribute to determining the wine typicity of the wine region.

Brewing a Craft Belgian-Style Pale Ale Using Pichia kudriavzevii 4A as a Starter Culture.

There is an expanding market for beer of different flavors. This study aimed to prepare a craft Belgian-style pale ale with a non-Saccharomyces yeast. Pichia kudriavzevii 4A was used as a sole starter culture, and malted barley as the only substrate. The ingredients and brewing process were carefully monitored to ensure the quality and innocuousness of the beverage. During fermentation, the yeast consumed 89.7% of total sugars and produced 13.8% v/v of ethanol. The product was fermented and then aged for 8 days, adjusted to 5% v/v alcohol, and analyzed. There were no traces of mycotoxins, lead, arsenic, methanol, or microbiological contamination that would compromise consumer health. According to the physicochemical analysis, the final ethanol concentration (5.2% v/v) and other characteristics complied with national and international guidelines. The ethyl acetate and isoamyl alcohol present are known to confer sweet and fruity flavors. The sensory test defined the beverage as refreshing and as having an apple and pear flavor, a banana aroma, and a good level of bitterness. The judges preferred it over a commercial reference sample of Belgian-style pale ale made from S. cerevisiae. Hence, P. kudriavzevii 4A has the potential for use in the beer industry.

The Impact of Yeast Encapsulation in Wort Fermentation and Beer Flavor Profile.

The food and beverage industry is constantly evolving, and consumers are increasingly searching for premium products that not only offer health benefits but a pleasant taste. A viable strategy to accomplish this is through the altering of sensory profiles through encapsulation of compounds with unique flavors. We used this approach here to examine how brewing in the presence of yeast cells encapsulated in alginate affected the sensory profile of beer wort. Initial tests were conducted for various combinations of sodium alginate and calcium chloride concentrations. Mechanical properties (i.e., breaking force and elasticity) and stability of the encapsulates were then considered to select the most reliable encapsulating formulation to conduct the corresponding alcoholic fermentations. Yeast cells were then encapsulated using 3% (w/v) alginate and 0.1 M calcium chloride as a reticulating agent. Fourteen-day fermentations with this encapsulating formulation involved a Pilsen malt-based wort and four S. cerevisiae strains, three commercially available and one locally isolated. The obtained beer was aged in an amber glass container for two weeks at 4 °C. The color, turbidity, taste, and flavor profile were measured and compared to similar commercially available products. Cell growth was monitored concurrently with fermentation, and the concentrations of ethanol, sugars, and organic acids in the samples were determined via high-performance liquid chromatography (HPLC). It was observed that encapsulation caused significant differences in the sensory profile between strains, as evidenced by marked changes in the astringency, geraniol, and capric acid aroma production. Three repeated batch experiments under the same conditions revealed that cell viability and mechanical properties decreased substantially, which might limit the reusability of encapsulates. In terms of ethanol production and substrate consumption, it was also observed that encapsulation improved the performance of the locally isolated strain.

Screening of Saccharomyces cerevisiae and Torulaspora delbrueckii strains in relation to their effect on malolactic fermentation.

The use of Torulaspora delbrueckii in the alcoholic fermentation (AF) of grape must is increasingly studied and used in the wine industry. In addition to the organoleptic improvement of wines, the synergy of this yeast species with the lactic acid bacterium Oenococcus oeni is an interesting field of study. In this work, 60 strain combinations were compared: 3 strains of Saccharomyces cerevisiae (Sc) and 4 strains of Torulaspora delbrueckii (Td) in sequential AF, and four strains of O. oeni (Oo) in malolactic fermentation (MLF). The objective was to describe the positive or negative relationships of these strains with the aim of finding the combination that ensures better MLF performance. In addition, a new synthetic grape must has been developed that allows the success of AF and subsequent MLF. Under these conditions, the Sc-K1 strain would be unsuitable for carrying out MLF unless there is prior inoculation with Td-Prelude, Td-Viniferm or Td-Zymaflore always with the Oo-VP41 combination. However, from all the trials performed, it appears that the combinations of sequential AF with Td-Prelude and Sc-QA23 or Sc-CLOS, followed by MLF with Oo-VP41, reflected a positive effect of T. delbrueckii compared to inoculation of Sc alone, such as a reduction in L-malic consumption time. In conclusion, the obtained results highlight the relevance of strain selection and yeast-LAB strain compatibility in wine fermentations. The study also reveals the positive effect on MLF of some T. delbrueckii strains.

Analysis of volatile compounds production kinetics: A study of the impact of nitrogen addition and temperature during alcoholic fermentation.

Among the different compounds present in the must, nitrogen is an essential nutrient for the management of fermentation kinetics, also playing a major role in the synthesis of fermentative aromas. Fermentation temperature is yet another variable that affects fermentation duration and the production of fermentative aromas in wine. The main objective of this study was thus to evaluate the combined effects of nitrogen addition-at the start of the fermentation process or during the stationary phase-at different fermentation temperatures on both fermentation kinetics and aroma synthesis kinetics. To study the impact of these three parameters simultaneously, we used an innovative transdisciplinary approach associating an online GC-MS system with an original modeling approach: a Box-Behnken experimental design combined with response surface modeling and GAM modeling. Our results indicated that all three factors studied had significant effects on fermentation and aroma production kinetics. These parameters did not impact in the same way the different families of volatile compounds. At first, obtained data showed that reduction of ester accumulation in the liquid phase at high temperature was mainly due to important losses by evaporation but also to modifications of yeast metabolic capabilities to synthetize these compounds. In a noticeable way, optimal temperature changed for liquid accumulation of the two classes of esters-23°C for acetate ester and 18°C for ethyl esters-because biological impact of temperature was different for the two chemical families. Moreover, the study of these three factors simultaneously allowed us to show that propanol is not only a marker of the presence of assimilable nitrogen in the medium but above all a marker of cellular activity. Finally, this work enabled us to gain a deeper understanding of yeast metabolism regulation. It also underlines the possibility to refine the organoleptic profile of a wine by targeting the ideal combination of fermentation temperature with initial and added nitrogen concentrations. Such observation was particularly true for isoamyl acetate for which interactions between the three factors were very strong.

Thiamine and Biotin: Relevance in the Production of Volatile and Non-Volatile Compounds during Saccharomyces cerevisiae Alcoholic Fermentation in Synthetic Grape Must.

Vitamins are major cofactors to numerous key metabolic pathways in enological yeasts, and both thiamine and biotin, notably, are believed to be essential to yeast fermentation and growth, respectively. In order to further assess and clarify their role in winemaking, and in the resulting wine, alcoholic fermentations of a commercial Saccharomyces cerevisiae active dried yeast were conducted in synthetic media containing various concentrations of both vitamins. Growth and fermentation kinetics were monitored and proved the essential character of biotin in yeast growth, and of thiamine in fermentation. The synthetic wine volatile compounds were quantified, and notable influences of both vitamins appeared, through a striking positive effect of thiamine on the production of higher alcohols, and of biotin on fatty acids. Beyond the evidence of this influence on fermentations and on the production of volatiles, this work proves, for the first time, the impact held by vitamins on wine yeasts' exometabolome, investigated through an untargeted metabolomic analysis. This highlighted chemical differences in the composition of synthetic wines through a notably marked influence of thiamine on 46 named S. cerevisiae metabolic pathways, and especially in amino acid-associated metabolic pathways. This provides, overall, the first evidence of the impact held by both vitamins on the wine.

Improving the Aromatic Profiles of Catarratto Wines: Impact of Metschnikowia pulcherrima and Glutathione-Rich Inactivated Yeasts.

Catarratto is one of the most widely cultivated grape varieties in Sicily. It is an indigenous non-aromatic white grape variety. Despite its widespread use in winemaking, knowledge of the aroma and chemical and microbiological properties of Catarratto wines is quite limited. The influence of Metschnikowia pulcherrima combined with Saccharomyces cerevisiae on the aromatic expression of Catarratto wines was investigated with and without the addition of glutathione-rich inactivated yeast. The substance is a natural specific inactivated yeast with a guaranteed glutathione level used to limit oxidative processes. The aromatic profiles of the final wines were determined through analysis of the volatile organic compounds using a solid-phase microextraction technique that identified 26 aromatic compounds. The addition of M. pulcherrima in combination with the natural antioxidant undoubtedly increased the aromatic complexity of the wines. Dodecanal was exclusively detected in the wines processed with glutathione-rich inactivated yeasts. Furthermore, the presence of this natural antioxidant increased the concentration of six esters above the perception threshold. Sensory analysis was also performed with a panel of trained judges who confirmed the aromatic differences among the wines. These results suggest the suitability of glutathione-rich inactivated yeasts for determining the oxidative stability of Catarratto wines, thus preserving its aromatic compounds and colour.

Redox cofactor metabolism in Saccharomyces cerevisiae and its impact on the production of alcoholic fermentation end-products.

The alcoholic fermentation of organic carbon sources by Saccharomyces cerevisiae produces many by-products, with the most abundant originating from central carbon metabolism. The production of these metabolites involves redox reactions and largely depends on the maintenance of redox homeostasis. Despite the metabolic pathways being mostly conserved across strains of S. cerevisiae, their production of various amounts of metabolic products suggests that their intracellular concentration of redox cofactors and/or redox balance differ. This study explored the redox status dynamics and NAD(H) and NADP(H) cofactor ratios throughout alcoholic fermentation in four S. cerevisiae strains that exhibit different carbon metabolic fluxes. This study focussed on the molecular end-products of fermentation, redox cofactor ratios and the impact thereof on redox homeostasis. Strain-dependent differences were identified in the redox cofactor levels, with NADP(H) ratios and levels remaining stable while NAD(H) levels decreased drastically as the fermentation progressed. Changes in the NAD+/NADH ratio were also observed. Total levels of NAD(H) decreased drastically as the fermentation progressed despite the cells remaining viable until the end of fermentation. NAD+ was found to be favoured initially while NADH was favoured towards the end of the fermentation. The change in the NAD+/NADH redox cofactor ratio during fermentation was linked with the production of end-products. The findings in this study could steer further research in the selection of S. cerevisiae wine strains for desirable aroma contributions based on their intracellular redox balance management.