The aim of the present study was to investigate the feasibly of using traditional milk kefir grains for the production of water kefir-like beverages and assess the changes in the physicochemical characteristics and the microbial populations of the fermented beverages. To this end, experiments of milk fermentation were primarily conducted at different temperatures and upon selection of the optimal, a gradual substitution of the substrate was performed by replacing milk from a sucrose-based solution. After the successful fermentation of the sucrose substrate, fruit juices were used as fermentation substrates. Sensory evaluation of the sugar-based beverages was also performed in order to access their acceptability for consumption. According to the results, the transition from milk to water kefir is indeed feasible, leading to the production of beverages with relatively higher ethanol concentrations (up to 2.14 ± 0.12% w/v) than milk kefir and much lower lactic acid concentrations (up to 0.16 ± 0.01% w/v). During the fermentation of the sugary substrates, yeasts seemed to be dominant over lactic acid bacteria, in contrast to what was observed in the case of milk kefir, where LAB dominated. The sensory evaluation revealed that all sugar-based beverages were acceptable for consumption, with the fruit-based ones obtaining, though, a better score in all attributes.