GNPS - Bile Salt Hydrolase Screening with bile acid and amines

120 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 18 bile acids and 49 amines. Following incubation, the reactions were quenched, and the extracts were dried and reconstituted in 80% methanol containing an internal standard. MS/MS data acquisition was carried out on Thermo Exploris 240 mass spectrometer in positive ionization mode, with chromatographic separation achieved using a Phenomenex Polar C18 column.