This study aimed to determine antioxidant activity, total phenol, phenolic acids and minimum inhibitory concentrations of the stem bark and root of three Barberries species (Berberis integrima Bunge, Berberis vulgaris asperma and Berberis vulgaris orientalis). The root extract of Berberis vulgaris asperma showed the highest amount of total phenol and antioxidant activity. After measuring minimum inhibition concentration, the stem extract of Berberis vulgaris orientalis and the root extract of Berberis vulgaris asperma were selected as the best extract with a high level of antimicrobial activity. Also, the root extract of Berberis vulgaris asperma could effectively prevent fungal and bacterial growth in a concentration of 600 and 750 ppm. Moreover, bread containing 600 ppm of Berberis vulgaris asperma root extract showed the highest level of overall acceptability and oxidative stability. According to the results, Berberis vulgaris asperma root extract was the best extract among the studied samples and the concentration of 600 ppm was the best level for enriching bread and producing functional bread with extended shelf life.