The goal of the project was to produce a functional probiotic fermented cold brewed coffee that is perapered with medium roasted, grinded Colombian Arabica beans with patented probiotic yeast Saccharomyces boulardii that can survive in human gastrointestinal conditions. With pasteurization of the product, safe fermentation environment and long shelf life was achieved. Coffee brew samples were prepared with different techniques to understand the effects of fermentation and glucose fortification to fermentation media on coffee chemistry, microbiota and sensory characteristic. One of the three coffee brews was only pasteurized (P), one of them fermented (F) and the other was fermented with glucose (FG). The cold brewing and fermentation were performed respectively at 22 °C and 30 °C for 24 hours. The brewed samples were pasteurized before fermentation at 65 °C for 30 minutes. Each sample was examined during their 120 days shelf life (at +4 °C). Although the probiotic yeast enabled fermentation in coffee, it did not survive after the first week. Pasteurization was successful, so pathogen growth was not detected in any of the samples during their shelf life. While FG was evaluated as undrinkable, the other samples gave positive results in terms of sensory. Caffeine, chlorogenic acid, and ethanol levels were significantly altered during shelf life. This is the first study in which chemical, microbiological and sensory analyzes of cold coffee that was brewed via cold immersion method, pasteurized and fermented with patented S. boulardii yeast during shelf-life were conducted, and will lead to new studies in this field.