Bacillus velezensis-derived laccase and superoxide dismutase detoxify aflatoxin B1, zearalenone, and alternariol

Aflatoxin B₁(AFB₁), zearalenone(ZEN), and alternariol(AOH) are major mycotoxins that threaten food safety, yet enzymes capable of detoxifying these structurally diverse toxins remain limited. In this study, two enzymes from Bacillus velezensis HNGD-JQ06, laccase BVLac and superoxide dismutase BV2SOD, were identified as broad-spectrum detoxification candidates. BVLac degraded 93.33% of AFB₁ at 60 °C and pH 7.0, 94.10% of ZEN at 60 °C and pH 8.0, and 96.87% of AOH at 50 °C and pH 8.0. BV2SOD degraded 92.59% of AFB₁ and 91.99% of ZEN at 50 °C and pH 7.0, and 98.94% of AOH at 60 °C and pH 8.0. UHPLC-MS/MS analysis suggested that AFB₁, ZEN, and AOH were transformed into products tentatively assigned as AFQ₁/epi-AFQ₁, 15-OH-ZEN, and a hydroxylated AOH derivative, respectively. Zebrafish assays confirmed reduced toxicity of the products, while molecular docking supported favorable enzyme-toxin binding. Both enzymes also retained activity in peanut, corn, and wheat flour matrices, highlighting their potential for multi-mycotoxin biodetoxification.