<title>Abstract</title> Vinegar production worldwide typically follows a natural fermentation process, which complicates control and standardization procedures. To streamline this process, it is essential to develop fermenting agents with high acetic acid production through acetic acid bacteria (AAB). The acetic acid present in the pollen of stingless bees is the most sensorially prominent organic acid, confirming that this bee product undergoes a fermentation process. This study aimed to isolate acetic acid bacteria from the pollen of Melipona subnitida bees, which are native to Brazil and whose products remain underexplored.. Thirteen strains were isolated from the pollen and one strain from kombucha as a control. The biochemical tests performed were: Gram staining, catalase production, H₂S production, indole production, motility, and catalase production. Additionally, an acetic acid production test was performed in culture medium and honey alcoholic fermentate. ANOVA and Tukey's test were applied. Finally, four strains were selected: MV1, MA2, PV6, and K, as promising for acetic acid production.