Improving aromatic higher alcohol acetates in wines by co‐fermentation of Pichia kluyveri and Saccharomyces cerevisiae: growth interaction and amino acid competition

Abstract BACKGROUND Higher alcohol acetates (HAAs) are potent aroma‐active esters that impart desirable fruity and floral aromas. However, the conversion of higher alcohol precursors into HAAs is extremely low in winemaking. To investigate the underlying yeast–yeast interaction on targeted improvement of aromatic HAAs, we evaluated fermentation activity, cell viability, amino acid consumption and HAA production when Pichia kluyveri and Saccharomyces cerevisiae were inoculated concurrently or sequentially. RESULTS Pichia kluyveri PK‐21 possessed the ability to survive and increased HAA level up to 5.2‐fold in mixed fermentation. Such an increment may benefit from the efficient conversion of higher alcohol precursors into HAAs (>27‐fold higher than S. cerevisiae ). During mixed fermentation, the two yeasts exhibited crucial interactions regarding cell growth and amino acid competition. Saccharomyces cerevisiae dominated over the co‐inoculated P. kluyveri by efficient uptake of amino acids and biomass production. However, this dominance decreased in sequential fermentation, where P. kluyveri growth increased due to the consumption of preferred amino acids prior to S. cerevisiae . Pearson correlation analysis indicated that phenylalanine and aspartic acid may act as positive amino acids in boosting P. kluyveri growth and HAA production. Laboratory‐scale winemaking validated the fermentation performance of P. kluyveri in sequential inoculum, resulting in a balanced aroma profile with enhanced floral and tropical fruity characteristics in the final wines. CONCLUSION This study proposes a microbial, non‐genetically engineered approach for targeted increase of HAA production in winemaking and the findings provide new insights into yeast–yeast interactions. © 2024 Society of Chemical Industry.