Kombucha is a Far East-originated fermented beverage that is valued by people for its health benefits worldwide. Fruit and fruit juice of the gilaburu plant is used in traditional medi-cine. In this study, fruit juice of gilaburu is used in a 21-day kombucha fermentation. Sensory properties, microbiological profile, total phenolic and flavonoid content, and antioxidant and antibacterial activity of the prepared beverages for days 0, 7, 14, and 21st were investigated. In addition, chemical compound profiles were determined using LC-MS/MS on the 7th and 14th days. When evaluated in general, gilaburu-flavoured kombucha yielded better results than traditional kombucha in terms of microbiological analysis, antibacterial activity, total phenolic content, and sensory properties. In addition, the LC-MS/MS analysis of the beverages revealed fourteen active compounds. Notably, on the 7th day, gilaburu-flavoured kombucha exhibited elevated levels of fumaric acid, quinic acid, chlorogenic acid, catechin, 4-OH-benzoic acid, epicatechin, vitexin and hesperidin compared to traditional kombucha.