The study evaluated the capability of *Bacillus subtilis* ZJ-2019-1 to simultaneously degrade aflatoxin B1 (AFB1) and zearalenone (ZEN), two potent mycotoxins that pose significant health risks to humans and animals. The strain was found to eliminate 60.88% of AFB1 and 33.18% of ZEN within 72 hours at a concentration of 10 mg/L at 37°C (pH 7.0), with enhanced activity under alkaline conditions. Autoclaved and boiled supernatants of *B. subtilis* ZJ-2019-1 significantly enhanced the degradation of both mycotoxins, achieving degradation rates of 79.85% and 100%, respectively, at a concentration of 1 mg/L within 48 hours at 37°C. Crude enzymes from *B. subtilis* ZJ-2019-1 showed maximum degradation rates for AFB1 (100%) and ZEN (94.29%) within 72 hours at 70°C. Divalent cations (such as Co2+, Fe2+, Mn2+, and Ni2+) significantly augmented the activity of crude enzymes. When applied to corn gluten meals, *B. subtilis* ZJ-2019-1 effectively detoxified 66.08% of AFB1 and 22.01% of ZEN, surpassing the efficacy of a commercial detoxification agent. These findings indicate that *B. subtilis* ZJ-2019-1 is a promising candidate for the simultaneous removal of multiple mycotoxins in food and feed.The study evaluated the capability of *Bacillus subtilis* ZJ-2019-1 to simultaneously degrade aflatoxin B1 (AFB1) and zearalenone (ZEN), two potent mycotoxins that pose significant health risks to humans and animals. The strain was found to eliminate 60.88% of AFB1 and 33.18% of ZEN within 72 hours at a concentration of 10 mg/L at 37°C (pH 7.0), with enhanced activity under alkaline conditions. Autoclaved and boiled supernatants of *B. subtilis* ZJ-2019-1 significantly enhanced the degradation of both mycotoxins, achieving degradation rates of 79.85% and 100%, respectively, at a concentration of 1 mg/L within 48 hours at 37°C. Crude enzymes from *B. subtilis* ZJ-2019-1 showed maximum degradation rates for AFB1 (100%) and ZEN (94.29%) within 72 hours at 70°C. Divalent cations (such as Co2+, Fe2+, Mn2+, and Ni2+) significantly augmented the activity of crude enzymes. When applied to corn gluten meals, *B. subtilis* ZJ-2019-1 effectively detoxified 66.08% of AFB1 and 22.01% of ZEN, surpassing the efficacy of a commercial detoxification agent. These findings indicate that *B. subtilis* ZJ-2019-1 is a promising candidate for the simultaneous removal of multiple mycotoxins in food and feed.