Metabolic Characterization of Two Flor Yeasts During Second Fermentation in the Bottle for Sparkling Wine Production

The global sparkling wine market continues to grow steadily, reaching approximately 24 million hectoliters in 2023, with an annual increase of around 4% despite a general decline in overall alcoholic beverage consumption. This growth highlights the importance of employing diverse yeast strains to improve product variety and quality. Flor yeasts are specialized strains of <i>Saccharomyces cerevisiae</i> that develop a biofilm on the surface of certain wines during biological ageing. They possess unique physiological properties, including high ethanol tolerance and the capacity to adhere, which supports wine clarification. They also have the ability to contribute unique volatile compounds and aroma profiles, making them promising candidates for sparkling wine production. This study evaluated two flor yeast strains (G1 and N62), which were isolated from the Pérez Barquero winery during the second fermentation process using the traditional method. Sparkling wines were produced by inoculating base wine (BW) with each strain, and the wines were monitored at 3 bar CO₂ pressure and after 9 months of ageing on lees. Comprehensive metabolomic analysis was performed using GC-MS for volatile compounds and HPLC for nitrogen compounds, with statistical analysis including PCA, ANOVA, Fisher's LSD, and correction FDR tests. Strain N62 demonstrated faster fermentation kinetics and higher cellular concentration, reaching 3 bar pressure in 27 days compared to 52 days for strain G1. Both strains achieved similar final pressures, 5.1-5.4 bars. Metabolomic profiling revealed significant differences in the profiles of volatile and nitrogen compounds between the two strains. G1 produced higher concentrations of 3-methyl-1-butanol, 2-methyl-1-butanol, and acetaldehyde, while N62 generated elevated levels of glycerol, ethyl esters, and amino acids, including glutamic acid, aspartic acid, and alanine. These findings demonstrate that both flor yeast strains successfully complete sparkling wine fermentation while producing distinct metabolic signatures that could contribute to unique sensory characteristics. This supports their potential as alternatives to conventional sparkling wine yeasts for enhanced product diversification.