Romain Minebois

The species Saccharomyces uvarum and Saccharomyces kudriavzevii have gained popularity in recent decades due to their interesting oenological properties. However, although it plays a crucial role in yeast fermentation performance and compound synthesis, our understanding of nitrogen metabolism in these species remains limited. Therefore, we compared how three strains of Saccharomyces cerevisiae , Saccharomyces uvarum and Saccharomyces kudriavzevii use relevant nitrogen sources by combining quantitative analysis approaches based on isotopic tracing and modelling. The model we have developed aims to facilitate the calculation and interpretation of stable isotope data for other experiments, by providing easy visualisation of the results and predicting the kinetics of isotope incorporation beyond the sampling points. The three species exhibit significant variations in their nitrogen assimilation profile. They differ in the timing of uptake of ammonium, arginine and glutamine: Saccharomyces cerevisiae prefers glutamine, Saccharomyces kudriavzevii ammonium and Saccharomyces uvarum arginine. This contributes to a different pattern of nitrogen redistribution towards proteinogenic amino acids between strains at the start of the exponential phase, which fades on entering the stationary phase. Additionally, we found that the contribution of leucine and valine to isoamyl alcohol production varies between species; also, Saccharomyces kudriavzevii activates the synthesis of volatile compounds earlier.

Nonconventional yeasts promise to provide new metabolic pathways for producing industrially relevant compounds and tolerating specific stressors such as cold temperatures. The mechanisms behind the cold tolerance of S. kudriavzevii or its sympatric relationship with S. cerevisiae in Mediterranean oaks are not well understood.

Saccharomyces non‐cerevisiae yeasts are gaining momentum in wine fermentation due to their potential to reduce ethanol content and achieve attractive aroma profiles. However, the design of the fermentation process for new species requires intensive experimentation. The use of mechanistic models could automate process design, yet to date, most fermentation models have focused on primary metabolism. Therefore, these models do not provide insight into the production of secondary metabolites essential for wine quality, such as aromas. In this work, we formulate a continuous model that accounts for the physiological status of yeast, that is, exponential growth, growth under nitrogen starvation and transition to stationary or decay phases. To do so, we assumed that nitrogen starvation is associated with carbohydrate accumulation and the induction of a set of transcriptional changes associated with the stationary phase. The model accurately described the dynamics of time series data for biomass and primary and secondary metabolites obtained for various yeast species in single culture fermentations. We also used the proposed model to explore different process designs, showing how the addition of nitrogen could affect the aromatic profile of wine. This study underlines the potential of incorporating yeast physiology into batch fermentation modelling and provides a new means of automating process design.