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A General Method for Detecting Nitrosamide Formation in the In Vitro Metabolism of Nitrosamines by Cytochrome P450s
Published on: September 25, 2017
A Novel Copper-Containing Nitrite Reductase for Simultaneous Detoxification of Aflatoxin B1 and Zearalenone:
Minjie Hou1, Shujie Zhang2, Yuhui Yang1
1College of Food Science and Engineering, Henan Key Laboratory of Cereal and Oil Food Safety and Nutrition, Henan University of Technology, Zhengzhou450001, P. R. China.
Abstract:
Co-contamination of aflatoxin B1 (AFB1) and zearalenone (ZEN) poses a major food safety risk. In this study, the copper-containing nitrite reductase AfNiR was identified as a novel biocatalyst for AFB1 and ZEN degradation, achieving maximum efficiencies of 99.22% and 93.53%, respectively, under optimal conditions of 40 °C, pH 7.0-8.0, 20 μg/mL (0.48 μM) enzyme dosage, and 24 h incubation for 5 μg/mL (16.01 μM) AFB1, and 50 °C, pH 8.0, 20 μg/mL (0.48 μM) enzyme dosage, and 24 h incubation for 10 μg/mL (31.41 μM) ZEN. UPLC-MS/MS identified AFQ1 and 15-OH-ZEN as the main transformation products, both exhibiting substantially reduced toxicity indicators in zebrafish assays. Molecular docking, site-directed mutagenesis, and molecular dynamics simulations indicated that both toxins bind near the T2 copper center and that copper-coordinating residues are essential for catalysis. To improve stability, AfNiR was immobilized in Cu3(PO4)2 nanoflowers and SA/SDS microspheres, achieving efficient detoxification in vegetable oil without affecting oil quality.
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