Formation Mechanism of Key Flavor Compounds During the Fermentation of Strawberry Juice with Water Kefir Grains

Water kefir grains are complex probiotic granules that can efficiently ferment fruit and vegetable juices and significantly improve product flavor. However, the mechanisms of flavor formation remain unclear, which limits the process optimization of this technology. This study investigated the mechanisms involved in flavor formation during the fermentation of strawberry juice with water kefir grains. The results showed that as fermentation progressed, the total acidity increased, whereas the pH value and soluble solids content decreased. Additionally, the contents of citric acid and malic acid gradually decreased with fermentation, while the contents of lactic, acetic, and succinic acid increased, and three soluble sugars showed reduced levels. A total of 218 volatile compounds were identified. Eight dominant bacterial genera and one dominant yeast species were detected. Significant correlations between some key microorganisms and flavor compounds were observed. Specifically, Lactiplantibacillus was positively correlated with hexyl acetate. Meanwhile, Gluconobacter and Acetobacter were positively correlated with methyl (Z,Z)-9,12-octadecadienoate, isoamyl acetate, etc. In contrast, LAB such as Lacticaseibacillus and Schleiferilactobacillus showed the opposite correlations with these key flavor compounds. Saccharomyces showed a positive correlation with ethyl palmitate, ethyl propionate, phenylsuccinic acid, and 1-pentanol. The main flavor compound metabolic pathways were predicted and they were significantly related with yeasts, acetic acid bacteria, and lactic acid bacteria. Overall, this study offers a theoretical basis for the directional regulation and optimization of the flavor quality of strawberry juice fermented with water kefir.