Molecular docking and mutation sites of CYP57A1 enzyme with Fomesafen

Fomesafen is a diphenyl ether herbicide developed by Zeneca Group PLC (UK), mainly used in soybean and peanut fields to control annual and perennial broad-leaved weeds. Fomesafen has strong persistence in the soil, slow degradation rate, and is prone to harm subsequent sensitive crops. This study utilized Autodock molecular docking technology to investigate the binding and interaction between degradation enzyme CYP57A1 and small molecules of fomesafen herbicides. The CYP57A1 gene cloned from a fomesafen-resistant fungus Fusarium verticilloids, belongs to a fragment of the P450 family, contains 587 bases, encodes 190 amino acids, and has an isoelectric point of 5.16. Visualization of the active surface of the protein receptor reveals that fomesafen is located in the cavity formed by the CYP57A1 protein and the cavity is small and tightly, the proteins are connected to small molecules through hydrogen bonds, halogen atom and π - cation interactions. Molecular modification of CYP57A1 enzyme was carried out using virtual amino acid mutation technology. Four key amino acids, LEU143, MET52, PHE176, and GLU177, were subjected to site-specific mutations. This study successfully constructed mutant engineered bacteria with stable protein expression. Mutations (1) MET52 > TRP showed a a decrease in enzyme activity, and the degradation rate of fomesafen was only 7.8 % of the wild-type. It is believed that MET52 is a key active site for the binding of CYP57A1 enzyme to small molecules of fomesafen, playing a crucial role in the degradation of fomesafen by this enzyme. This provides new insights into the impact on the degradation activity of fomesafen.