Modulation of Kombucha Functionality by Whey Protein-Encapsulated Lactobacillus: Effects on Bioactive Properties

Kombucha is a fermented beverage produced using a symbiotic consortium of acetic acid bacteria and yeasts, often marketed for its health-promoting properties. However, probiotic bacteria in kombucha are typically present at inconsistent levels and may not remain viable during fermentation. In this study, three Lactobacillus strains (Lacticaseibacillus rhamnosus ATCC 53103 (L. rhamnosus), Lactiplantibacillus plantarum subsp. plantarum ATCC 14917 (L. plantarum) and Lentilactobacillus hilgardii (L. hilgardii) isolate) were encapsulated in whey protein using the lyophilization method and added individually at the start of kombucha fermentation. Lactic acid bacteria (LAB)–enriched kombucha samples were evaluated for chemical composition (polyphenols, flavonoids, vitamin C and organic acids) and functional properties (antimicrobial, antiproliferative, antioxidant and anti-inflammatory activities) and compared to a traditionally obtained control kombucha, primarily demonstrating in vitro and experimental assessment. Encapsulation maintained LAB viability above 6–7 log CFU/mL throughout fermentation, producing kombucha with enhanced microbial stability. LAB–enriched samples exhibited increased L-lactic acid and antimicrobial activity. L. rhamnosus and L. hilgardii–enriched samples exhibited increased antiproliferative and anti-inflammatory activities, which may be associated with strain-dependent production of organic acids, polyphenol modulation and LAB-derived bioactive metabolites. Antioxidant activity varied depending on assay, and L. rhamnosus–enriched kombucha showed higher anti-inflammatory activity. These findings demonstrate that whey protein encapsulation can preserve LAB during fermentation, enhance specific bioactive properties and provide a platform for developing functional kombucha beverages with potential applications in the food industry.