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Modified attapulgite-laccase composites for aflatoxin B1 detoxification: Performance, toxicity, and mechanistic
1Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Laboratory of Quality and Safety Risk Assessment for Oilseeds Products (Wuhan), Quality Inspection and Test Center for Oilseeds Products, Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs, National Reference Laboratory for Agricultural Testing (Biotoxin), Wuhan 430062, PR China.
Abstract:
Aflatoxin B1 (AFB1) poses significant threats to both health and the economy. Recently, integrated technologies have emerged as a novel detoxification strategy. However, most current research remains at the laboratory stage, impeded by high costs, unknown toxicity of degradation products, and complex, time-consuming processes. In this study, a promising adsorption-biocatalytic system for AFB1 detoxification using engineered attapulgite-laccase composites (L-AATP) was constructed. L-AATP exhibited greater stability than laccase. Moreover, it displayed a strong detoxification ability, achieving a detoxification efficiency of 670 ng/mg. The 13 putative AFB1 degradation products were tentatively assigned by LC-MS, and three potential degradation pathways were proposed with support from density functional theory (DFT) calculations. Theoretical calculations suggested that the furan ring double bond may be a susceptible site for electrophilic addition, while lactone-ring hydrolysis and opening were also thermodynamically feasible. Toxicity assessment using the T.E.S.T. program predicted that most degradation intermediates generated during L-AATP-mediated detoxification may have lower toxicity than AFB1. An in vitro viability assay demonstrated low cytotoxicity of L-AATP to IPEC-J2 cells. To assess the potential applicability of immobilized laccase, its performance was evaluated in contaminated peanut meal under simulated gastrointestinal conditions. A straightforward detoxification procedure was established: blending L-AATP with contaminated peanut meals at a mass ratio of 1:1000, which was appropriate when contamination levels do not surpass 300 μg/kg. In addition, L-AATP was an easy-to-operate, low-cost, long-term effective detoxifying agent, with negligible nutritional impact on peanut meals. This work not only developed a novel mycotoxin detoxifier but also provides new insights into the joint application of multiple technologies to enhance detoxification efficacy.
