The various forms of interactions that microorganisms engage in within fermented foods result in distinct metabolic product patterns. Fermentation products often feature both yeasts and bacteria, each possessing characteristics that can enhance the overall quality of the food, thus benefiting consumers. Kefir, a fermented milk originating from grains containing a unique and intricate blend of bacteria and yeasts living in a symbiotic relationship, is a valuable model for studying the evolution of the interactions between yeasts and bacteria. Targeted metagenomics was applied to investigate the microbiome of a batch of traditional Romanian kefir and further examine the growth and metabolic properties of the dominant yeast and bacterial strains isolated from this batch. In contrast to yeast, which is either unaffected or harmed by the presence of bacteria, our study revealed that a specific strain of Lactobacillus (L. rhamnosus) derived from the kefir batch benefited from the presence of Saccharomyces cerevisiae. The analysis of short-chain fatty acids (SCFAs) produced by in vitro cultures of these two chosen strains indicated significant changes in SCFA levels compared to single cultures. The dynamic interactions described in this and other studies on kefir emphasize the importance of a more profound comprehension of the ecological mechanisms governing interactions between yeast, bacterial, and mammalian cells.