Nitrogenous Compound Utilization and Production of Volatile Organic Compounds among Commercial Wine Yeasts Highlight Strain-Specific Metabolic Diversity

Abstract

ABSTRACTStrain and environmental nutrient concentrations can affect the production of sensory impact compounds during yeast fermentation. Despite reports on the impact of nutrient conditions on kinetics of cellular growth, it is uncertain to what extent nitrogen utilization by commercial Saccharomyces cerevisiae wine strains affects the production of volatile organic (aroma) compounds (VOCs). Here we ask whether i) consumption of amino acids contribute to VOCs (fusel alcohols, acetate esters, and fatty acid esters) in commercial S. cerevisiae yeast strains, ii) there is inter-strain variation in VOC production, and iii) there is a correlation between the production of aroma compounds and nitrogen utilization. We analyzed the consumption of nutrients as well as the production of major VOCs during fermentation of a chemically defined grape juice medium with four commercial S. cerevisiae yeast strains: Elixir, Opale, R2, and Uvaferm. The production of VOCs was variable among the strains where Uvaferm correlated with ethyl acetate and ethyl hexanoate production, R2 negatively correlated with the acetate esters, and Opale positively correlated with fusel alcohols. The four strains’ total biomass formation was similar, pointing to metabolic differences in the utilization of nutrients to form secondary metabolites such as VOCs. To understand the strain-dependent differences in VOC production, partial least-squares linear regression coupled with genome-scale metabolic modeling was performed with the objective to correlate nitrogen utilization with fermentation biomass and volatile formation. Total aroma production was found to be a strong function of nitrogen utilization (R2 = 0.87). We found that glycine, tyrosine, leucine, and lysine utilization were positively correlated with fusel alcohols and acetate esters concentrations e.g., 2-phenyl acetate during wine fermentation. Parsimonious flux balance analysis and flux enrichment analysis confirmed the usage of these nitrogen utilization pathways based on the strains’ VOC production phenotype.IMPORTANCESaccharomyces cerevisiae is widely used in grape juice fermentation to produce wines. Along with the genetic background, the nitrogen in the environment in which S. cerevisiae grows impacts its regulation of metabolism. Also, commercial S. cerevisiae strains exhibit immense diversity in their formation of aromas, and a desirable aroma bouquet is an essential characteristic for wines. Since nitrogen affects aroma formation in wines, it is essential to know the extent of this connection and how it leads to strain-dependent aroma profiles in wines. We evaluated the differences in the production of key aroma compounds among four commercial wine strains. Moreover, we analyzed the role of nitrogen utilization on the formation of various aroma compounds. This work illustrates the unique aroma producing differences among industrial yeast strains and suggests more intricate, nitrogen associated routes influencing those aroma producing differences.