Development of a Dihydropteroate Synthase-Based Fluorescence Polarization Assay for Detection of Sulfonamides and Studying Its Recognition Mechanism.

In this study, the dihydropteroate synthase of Staphylococcus aureus was obtained, and its recognition mechanisms for 31 sulfonamide drugs were studied. Results showed that their core structure matched well with the binding pocket of para-aminobenzoic acid, and all the sulfonamide side chains were out of the binding pocket. Hydrogen bonds and hydrophobic interactions were the main intermolecular forces, and the key amino acids were Gly171 and Lys203. The binding sites in sulfonamide molecules were mainly around the para-aminobenzenesulfonamide part. This enzyme was used to develop a fluorescence polarization assay for detection of these drugs in chicken muscles. The change trends of half of inhibition concentrations and cross-reactivities for the 31 drugs were identical with the receptor-ligand affinities. The limits of detection were in the range of 2.0-38.5 ng/g, and one assay could be finished within several minutes. Therefore, this method could be used for multiscreening of sulfonamide residues in meat samples.