Flocculation Mechanisms in Brettanomyces bruxellensis: Influence of ethanol and sulfur dioxide on FLO gene expression

Abstract

The mechanisms underlying flocculation in Brettanomyces bruxellensis, unlike the well-characterized FLO-family gene regulation in Saccharomyces cerevisiae, remain largely unexplored. This study investigates the flocculant phenotypes of 99 B. bruxellensis strains, revealing that only a minority exhibits this clumping behavior and confirms its strain-dependent attitude. Focusing on two strains, CBS2499 (flocculant) and UMY321 (non-flocculant), genetic analysis uncovered polymorphisms and distinct allelic heterozygosity in the FLO1 and FLO11 genes, potentially linked to the phenotypic differences. To further examine these traits, Response Surface Methodology (RSM) was used to simulate oenological conditions, testing the impact of pH, ethanol, and sulfur dioxide (SO₂) levels on flocculation and gene expression. The findings revealed that environmental stressors, especially ethanol and SO₂, significantly increase the expression of FLO1 and FLO11 in CBS2499, indicating a regulatory role in flocculation under stress. These insights broaden our understanding of stress adaptation in B. bruxellensis, especially its survival strategies in wine environments. By elucidating factors influencing flocculation, this study contributes valuable knowledge for managing B. bruxellensis spoilage, potentially aiding in the development of targeted approaches to reduce its impact on wine quality.