Dual Substrate Catalytic Detoxification of Aflatoxin B1 and Zearalenone by a Novel Enzyme PsCopA from Pseudomonas for Food and Feed Safety

Fungal mycotoxins pose a serious threat to human and animal health, especially due to their co-occurrence in food and feed, causing synergistic toxicity. Current enzymatic detoxification strategies often depend on redox mediators, limiting enzyme diversity and restricting practical application. In this study, we characterized PsCopA, a novel multicopper oxidase from Pseudomonas sp., heterologously expressed in Escherichia coli , which efficiently degraded aflatoxin B 1 and zearalenone simultaneously without mediators. PsCopA showed optimal activity at 50°C (pH 7.0) for AFB 1 and 60°C (pH 8.0) for ZEN and retained over 80% activity after 2 h incubation across 20-80°C. UPLC-MS/MS analysis identified AFQ 1 and β-zearalanol as degradation products, both exhibiting significantly reduced hepatotoxicity and oxidative stress in zebrafish, even under co-exposure, suggesting effective mitigation of synergistic toxicity. Notably, PsCopA removed over 90% of toxins from contaminated cornmeal and peanut meal. These findings demonstrate PsCopA has a promising prospect for industrial application in food and feed detoxification.