GNPS_Bile_Salt_Hydrolase_screening_pilot_exploris

Bile Salt Hydrolases (BSH) and Penicillin V Acylase (PVA) enzymes from various microbial strains were overexpressed in E. coli and purified using nickel affinity chromatography. Enzyme assays were performed by incubating the purified enzymes with bile acids and amines. Following incubation, the reactions were quenched, and the extracts were dried and reconstituted in 50% methanol containing an internal standard. MS/MS data acquisition was carried out on a Thermo Exploris mass spectrometer in positive ionization mode, with chromatographic separation achieved using a Phenomenex Polar C18 column.