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.