Xing_2025_Food.Chem_500_147404

An acetylcholinesterase from Aspergillus niger GZUF36 (AnAChE) was expressed in Escherichia coli BL21(DE3) and comprehensively characterized with respect to its biochemical properties and suitability for pesticide analysis. The recombinant enzyme (rAnAChE) exhibited typical AChE substrate specificity, with optimal activity at pH 7.0 and 30-35 degreesC. rAnAChE tolerated hydrophobic solvents and methanol, supporting its use in practical assays. The enzyme was highly sensitive to methomyl, carbofuran, trichlorfon, fenitrothion, and carbaryl. For the 12 pesticides tested, the limits of detection of most pesticides in simple matrices such as rice and spinach were below China's maximum residue limits (GB 2763-2021), indicating the enzyme has strong potential for pesticide monitoring. Molecular docking and dynamics simulations revealed competitive binding modes and identified Leu190 and Tyr269 as key recognition residues. Overall, this work provides a promising enzymatic tool for pesticide monitoring and elucidates structure-function relationship of AnAChE, contributing to food safety and environmental protection.