Phylogenomics and functional annotation of 530 non-Saccharomyces yeasts from winemaking environments reveals their fermentome and flavorome.

The winemaking industry faces unprecedented challenges due to climate change and market shifts, with profound commercial and socioeconomic repercussions. In response, non-Saccharomyces yeasts have gained attention for their potential to both mitigate these challenges and enhance the complexity of winemaking. This study builds upon our previous cataloguing of 293 non-Saccharomyces yeast species associated with winemaking environments by rigorously analysing 661 publicly available genomes. By employing a bioinformatics pipeline with stringent quality control checkpoints, we annotated and evaluated these genomes, culminating in a robust dataset of 530 non-Saccharomyces proteomes, belonging to 134 species, accessible to the research community. Employing this dataset, we conducted a comparative phylogenomic analysis to decipher metabolic networks related to fermentation capacity and flavor/aroma modulation. Our functional annotation has uncovered distinctive metabolic traits of non-Saccharomyces yeasts, elucidating their unique contributions to enology. Crucially, this work pioneers the identification of a non-Saccharomyces 'fermentome', a specific set of six genes uniquely present in fermentative species and absent in non-fermentative ones, and an expanded set of 35 genes constituting the complete fermentome. Moreover, we delineated a 'flavorome' by examining 96 genes across 19 metabolic categories implicated in wine aroma and flavour enhancement. These discoveries provide valuable genomic insights, offering new avenues for innovative winemaking practices and research. Citation: Franco-Duarte R, Fernandes T, Sousa MJ, Sampaio P, Rito T, Soares P (2025). Phylogenomics and functional annotation of 530 non-Saccharomyces yeasts from winemaking environments reveals their fermentome and flavorome. Studies in Mycology 111: 1-17. doi: 10.3114/sim.2025.111.01.