Kefir drink represents a functional product, with great nutritive value, made by fermentation of sugary medium through 2-4 days at room temperature by consortium of microorganisms contained in water kefir grains. It also has proven probiotic properties, which has antihyperglycemic, antimutagenic, and antioxidative effects of human body. Although kefir drink has many positive properties, its industrial production is still very limited. Varying fermentation conditions, and poorly known consortium of microorganisms in water kefir grains makes the process of kefir drink production very hard to precisely define. The breakthrough in industrial production of kefir drink is possible only if the process parameters are well known. Different substrates could also have a great impact on kefir drink production, and they could also give better sensoric properties to the final product. In this study, the optimum temperature and substrate concentrations were defined (25 ℃ / 60 g L-1). Cane sugar was the main source of sucrose, which was used to make the sugary medium, which was later sterilised and inoculated with 10 g of kefir drink grains. The effects of sweeteners (erythrol, xylitol and steviol) on production of kefir drink on cane were studied, where there were no negative effects of sweeteners on the bioprocess itself discovered, which makes them eligible to use after, but also during the fermentation process.