Global trends indicate a growing interest in foods with higher nutritional value and functional properties, which encourages the food industry to develop health-promoting products. In this context, fermented foods are gaining importance due to their high content of functional molecules. In our research, we investigated the lactic acid fermentation of a mixture of honey, bee pollen, and royal jelly (API mixture) to enhance its functional value. The L. plantarum culture proved to be the most suitable for fermentation, while L. brevis and L. fermentum were less effective. Optimization of the fermentation conditions showed that a 1% inoculum was sufficient to initiate fermentation. Dilution ratios of 1:5 and 1:10 were found to be suitable, while 1:1 and 1:2 were less effective due to inhibition. During fermentation, the number of microorganisms (CFU/mL) increased and the pH value decreased, with the most significant changes occurring at the beggining of the fermentation process, followed by stabilization. We observed a smaller decrease in the glucose content (25.4% decrease after 72 hours), while the fructose content did not change significantly. The organic acid content, particularly lactic and acetic acid, increased during fermentation. An increase in total phenolic compounds, 10-HDA or kynurenic acid was not achieved. To further improve fermentation, we used a phosphate buffer, which contributed to a more stable pH during fermentation and a greater decrease in glucose content (36.4 % decrease after 72 hours) and in some cases fructose content (13.9% decrease after 72 hours). These results indicate the potential of the fermented API mixture as a dietary supplement, but further research is needed, particularly on the metabolites and on improving the conditions for lactic acid fermentation.
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