In this study, peroxidase (POD) and laccase (LC) were selected to enzymatically degrade zearalenone (ZEN). Reaction parameters, including enzyme concentration, pH, and temperature, were optimized, and kinetic models were established to evaluate their catalytic characteristics. Additionally, the degradation efficiencies of ZEN in beer as a food matrix by POD and LC were assessed. The results showed that under the optimal reaction conditions of 40 ℃, pH 7, and 48 U/mL, POD degraded more than 90% of ZEN. When activated by 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), methyl syringate (MSG), or acetosyringone (AS), LC could also degrade more than 90% of ZEN. Kinetic analysis indicated that both POD and LC exhibited interactions with ZEN. The K_M and V_max values for POD were 2.01 μg/mL and 0.015 μg/(mL·min), respectively. The K_M and V_max values for LC were 2.66 μg/mL and 0.017 μg/(mL·min), respectively. Compared with model solutions, the degradation efficiencies of ZEN in beer were reduced due to acidic pH, metal ions, and matrix complexity. This study provides a theoretical basis for enzymatic detoxification of mycotoxins in food systems and supports the development of functional enzyme preparations.

Zearalenone (ZEN), a mycotoxin present in cereals, poses significant health risks to animals and humans due to its estrogenic effects. Numerous studies on the enzymatic detoxification of ZEN have predominantly focused on reducing the parent toxin to assess the enzyme’s efficacy, yet there is limited research on the identification and toxicity evaluation of the enzymatic degradation products. This study investigated the enzymatic degradation mechanisms of ZEN using commercial peroxidase (POD) and laccase (LC), with a focus on identifying degradation products and assessing their hepatotoxicity effects. Molecular docking and dynamics simulations elucidated the binding mechanisms between these enzymes and ZEN, revealing strong interactions that facilitate efficient detoxification. Subsequent analysis employing ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) successfully identified crucial degradation products. Hepatic toxicity of the enzymatic degradation products was comprehensively assessed in HepaRG liver cells through systematic measurements of cell viability, oxidative stress, apoptosis, mitochondrial membrane potential, and molecular metabolic profiles. Our findings demonstrate that both POD and LC exhibit significant efficacy in mitigating hepatocyte toxicity induced by ZEN, thereby highlighting their potential utility in enhancing food safety. This research provides essential data for safety evaluation regarding enzymatic detoxification of ZEN while offering theoretical and technical resources for risk assessment related to mycotoxin enzymatic detoxification.