Abstract 1059 Optimizing Microfluidic Gradient Generating SPR Chips for High-Throughput Biochemical Reaction Kinetics in Renal Physiology

The esterases GloB and FrmB from Staphylococcus aureus are pivotal in activating carboxy ester prodrugs.This study aims to elucidate the structures of GloB and FrmB while bound to specific ligands to enhance our understanding of their enzymatic mechanisms and potential therapeutic applications.The primary objective was to determine the crystal structures of GloB and FrmB in complex with ligands using X-ray crystallography.We expressed, purified, and crystallized GloB and FrmB, in preparation for X-ray crystallography to resolve their structures.Various crystallization conditions were tested to optimize crystal formation.Successful crystal formation was achieved primarily under conditions with PEG 3350 compared to Tris pH 7.5 buffer, which provided clusters of needles.Crystals were harvested and high-resolution structures will be obtained at the AMX beam line at the NSLS2 synchrotron, which will reveal detailed interactions between the esterase and their bound ligands.