This study examined the impact of Limosilactobacillus fermentum NCU001464 fermentation on physicochemical properties, xanthine oxidase inhibitory activity, and the flavor profile of Pueraria Lobata (PL). An orthogonal experiment determined the optimum conditions: inoculation amount of 6 %, a fermentation time of 32 h, and a fermentation temperature of 37 °C. Under these conditions, fermented PL exhibited significantly higher antioxidant capacity and xanthine oxidase inhibition ability than unfermented PL, possibly because of increased total flavonoid content from 1.27 mg RUT/mL to 1.50 mg RUT/mL. Notably, The compounds, kaempferol 3-rhamnoside 4'-xyloside and puerarin xyloside, significantly increased after fermentation, and molecular docking and dynamic simulations revealed their strong interactions with xanthine oxidase. Additionally, fermentation improved the flavor by reducing the inferior aroma of fat and green and increasing the sweet fruity notes. These results highlight the potential of fermented PL as a functional food and provide valuable theoretical foundations for its high-value applications.