Arbutin is a combination of D-glucose and hydroquinone. Among the positive effects of arbutin are its antioxidant properties, antimicrobial properties and anti-inflammatory properties. Arbutin is one of the most important active compounds used to inhibit the enzyme tyrosinase, which causes enzymatic browning in many foods. Microbial fermentation has emerged as an up-and-coming technique for the production of arbutin in recent years. The aim of the study was the biological production of arbutin from Bacillus subtilis NN2M isolates. The isolates were confirmed and identified by Vitek and 16S rRNA and the nutritional conditions for production were improved to reach optimal conditions. To determine the best nutritional conditions for arbutin production, several conditions were studied, including the carbon source and its concentration, the nitrogen source and its concentration and the hydroquinone concentration. The results showed that the highest output of the active compound arbutin when using Molasses as a carbon source arbutin concentration was 23.34µg/mL. We also found the best concentration of the optimal carbon source, 4 % (w\v) arbutin concentration, was 27.62 µg/mL; furthermore, while using several nitrogen sources for producing arbutin, peptone was determined to be the optimum source, the concentration of arbutin was 47.3 µg/mL, the nitrogen source optimal Concentration was 2 % (w\v) arbutin concentration was 59.46 µg/mL, the optimum concentrations of hydroquinone was 2 % (w\v) arbutin concentration was 79.1 µg/mL.