Co-Cultivation of Potential Probiotic Strains Isolated from Water Kefir for Fermented Green Tea Beverage

This study aimed to isolate and characterize microorganisms from water kefir beverage for their functional properties. Five lactic acid bacteria (LAB) strains were isolated: three Leuconostoc citreum strains (LB4, LB6, LB13) and two Lactococcus lactis strains (LB5, LB25), identified via 16S rRNA sequencing, along with three Saccharomyces cerevisiae yeast strains (Y7, Y9, Y10), confirmed by 18S rDNA sequencing. Due to the high genetic and phenotypic similarity within each species, one representative strain from each (LB4, LB5, Y9) was selected for further analysis. These strains showed potential probiotic properties, including tolerance to acid and bile, high auto-aggregation, and hydrophobicity. The LAB strains were sensitive to gentamicin, and their supernatants inhibited the growth of tested pathogenic bacteria. The cumulative probiotic potential (CPP) scores were 93.33% for Lc. citreum LB4 and L. lactis LB5, and 100% for S. cerevisiae Y9. Furthermore, the fermentation potential of these strains was evaluated in a green tea beverage using three co-culture formulations. Among the formulations tested, the BF1 beverage, fermented by F1 (40% LB4, 40% LB5, and 20% Y9), demonstrated optimal physicochemical, microbiological, and sensory properties. Notably, while the individual strains did not show anti-inflammatory activity, the BF1 beverage formulation exhibited this effect, suggesting a synergistic interaction during fermentation.