Nicotinic acid deficiency impacts wine fermentation efficiency in Saccharomyces cerevisiae

This article is adapted from a version previously published in IVES Technical Reviews.

Article reference: Duncan, J.D. & Divol, B., 2025. Nicotinic acid deficiency impacts wine fermentation efficiency in Saccharomyces cerevisiae. IVES Technical Reviews. https://doi.org/10.20870/IVES-TR.2025.9271.

Nicotinic acid (NA), a form of vitamin B3, is vital for producing the essential redox cofactors NAD⁺ and NADP⁺. These cofactors drive many reactions required for yeast growth and fermentation. Since Saccharomyces cerevisiae cannot synthesise these cofactors under anaerobic conditions without external vitamin B3 in the must, the NA levels in grape juice become crucial. This study examined how NA deficiency affects yeast fermentation and the resulting metabolic profile of wine.

Nicotinic acid deficiency negatively impacts alcoholic fermentation kinetics

Researchers studied how different amounts of NA affect fermentation by Saccharomyces cerevisiae EC1118™ (Lallemand Inc., QC, Canada) in grape juice containing 230 g/L sugar (half glucose, half fructose) at 25°C. They tested three levels of NA: low (0.1 mg/L), normal (2.0 mg/L), and high (4.0 mg/L). In the first 24 hours, all fermentations progressed similarly. Thereafter, low NA caused a slower fermentation and a decrease in yeast growth by 37.5%. Fermentations at standard and high NA concentrations completed normally, confirming that a lack of NA impaired performance.

Other yeast species reacted differently. In Kluyveromyces marxianus and Lachancea thermotolerans, low NA slowed the fermentation even further and changed the amount of metabolites produced. In Starmerella bacillaris, fermentation initially progressed well, but later metabolic activity decreased, resulting in stuck fermentation. These findings highlight that yeast species respond differently to NA limitation, which can shape fermentation performance and the types of metabolites produced.

Deficient nicotinic acid results in an imbalance in redox cofactors

The redox cofactors NAD⁺ and NADH were quantified in the yeast cells throughout the fermentation. NAD⁺ levels initially rose until the end of the lag phase, then dropped sharply as cells entered exponential growth, reflecting high metabolic activity and rapid reduction of NAD⁺ to NADH. Ethanol, glycerol, and other metabolites are produced to help balance oxidised and reduced cofactors. However, during anaerobiosis, the NADH reoxidation cannot fully keep up, and the limited source of extracellular B3 subsequently results in the decline of both NAD⁺ and NADH levels after 24 hours. At this stage in the low NA treatment (0.1 mg/L), NAD⁺ became almost undetectable, creating a severe redox imbalance in the cell. This shortage of NAD⁺ slows fermentation, reduces biomass, and disrupts key metabolic processes, explaining the sluggish fermentations in S. cerevisiae and early stuck fermentations in some non-Saccharomyces yeasts.

Dynamics of nicotinic acid uptake and metabolism

At the start of fermentation, yeasts rely on their own vitamin B3 reserves to make NAD⁺, as well as importing NA from the grape juice. This consumption from the must peaks before the mid-point of yeast growth, when all available extracellular NA is assimilated. Thereafter, the cells recycle NAD⁺ into various forms of vitamin B3 within the cell, with a small amount of the vitamin exported back into the medium. Both NA uptake and export increased proportionally with the higher initial NA concentrations tested.

Deficiency impacts the production of major metabolites

The study measured how NA levels affect yeast metabolite production in synthetic grape juice. NA deficiency (0.1 mg/L) led to increased levels of glycerol, malic acid, acetoin and succinic acid, but decreased acetic acid, while ethanol production remained unchanged, as shown in Figure 1. Standard (2.0 mg/L) and high (4.0 mg/L) levels of NA resulted in similar metabolite production, indicating a threshold beyond which extra NA provides no benefit. This highlights the importance of sufficient vitamin B3 for maintaining redox balance, efficient fermentation, and consistent metabolite profiles.

FIGURE 1.The effects of nicotinic acid deficiency and sufficiency on yeast metabolism and fermentation dynamics.

When do deficiencies occur in real grape juice?

Vitamin B3 deficiency in grape juice is less common than vitamin B1 (thiamine) deficiency, but certain conditions can reduce its levels. Fungal infections of the berry, such as Botrytis cinerea, and yeast assimilation during early fermentation can lower NA concentrations. Extended pre-fermentative soaking or inoculating non-Saccharomyces yeasts may further increase the risk of deficiency. To prevent this, supplementation with yeast-derived nutrients containing vitamin B3 can be effective. Controlled oxygen addition can also help, as it enables the de novo biosynthesis of vitamin B3 by the yeast. However, the timing and dosage of oxygen must be carefully managed to support vitamin B3 production without leading to oxidation of the grape juice.

Conclusion

This study shows that NA plays a central role in yeast redox balance and metabolism in S. cerevisiae by supporting NAD⁺ regeneration and efficient fermentation. Low NA causes NAD⁺ depletion, slowing fermentation and reducing yeast growth, and altering the metabolite profile of the wine. An excess of NA offers no extra benefit, indicating an optimal supplementation level. Non-Saccharomyces yeasts respond differently to NA deficiency, with each influencing the fermentation profile differently. Future work should focus on optimising nutrient supplementation and oxygenation to support NAD⁺ production and maintain smooth, efficient fermentations, particularly in mixed-yeast fermentations.

Highlights

• Nicotinic acid (vitamin B3) is required for redox cofactor synthesis in yeasts.
• Low NA causes redox imbalance, leading to altered metabolite production and sluggish fermentations.
• Excess NA offers no benefit beyond normal levels (~2 mg/L).
• Some non-Saccharomyces yeasts are more sensitive to NA deficiency.
• Vitamin B3 supplementation or controlled oxygenation can prevent fermentation problems.

References

Duncan, J.D., Setati, M.E. & Divol, B., 2024. Nicotinic acid availability impacts redox cofactor metabolism in Saccharomyces cerevisiae during alcoholic fermentation. FEMS Yeast Research, 24, foae015. https://doi.org/10.1093/femsyr/foae015.

Evers, M.S., Roullier‐Gall, C., Morge, C., Sparrow, C., Gobert, A. & Alexandre, H., 2021. Vitamins in wine: Which, what for, and how much? Comprehensive Reviews in Food Science and Food Safety, 20(3), 2991-3035. https://doi.org/10.1111/1541-4337.12743.

Tyibilika, V., Setati, M.E., Bloem, A., Divol, B. & Camarasa, C., 2024. Exploring fermentative metabolic response to varying exogenous supplies of redox cofactor precursors in selected wine yeast species. FEMS Yeast Research, 24, foae029. https://doi.org/10.1093/femsyr/foae029.

Duncan, J.D., Setati, M.E. & Divol, B., 2023. Redox cofactor metabolism in Saccharomyces cerevisiae and its impact on the production of alcoholic fermentation end-products. Food Research International, 163, 112276. https://doi.org/10.1016/j.foodres.2022.112276.

Kelly, J., Inglis, D., Dowling, L. & Pickering, G., 2022. Impact of Botrytis cinerea‐infected grapes on quality parameters of red wine made from withered grapes. Australian Journal of Grape and Wine Research, 28(3), 439-449. https://doi.org/10.1111/ajgw.12545.

Bataillon, M., Rico, A., Sablayrolles, J.M., Salmon, J.M. & Barre, P., 1996. Early thiamin assimilation by yeasts under enological conditions: impact on alcoholic fermentation kinetics. Journal of Fermentation and Bioengineering, 82(2), 145-150. https://doi.org/10.1016/0922-338X(96)85037-9.

Labuschagne, P.W., Rollero, S. & Divol, B., 2021. Comparative uptake of exogenous thiamine and subsequent metabolic footprint in Saccharomyces cerevisiae and Kluyveromyces marxianus under simulated oenological conditions. International Journal of Food Microbiology, 354, 109206. https://doi.org/10.1016/j.ijfoodmicro.2021.109206.

For more information, contact Benoit Divol at [email protected].

Click here to get your copy of WineLand Magazine.