High-throughput screening assay for D-amino acid oxidase
Related Experiment Video
Updated: Jul 8, 2026
A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
Published on: August 17, 2019
Svetlana V Khoronenkova1, Vladimir I Tishkov
1Department of Chemical Enzymology, Chemistry Faculty, M. V. Lomonosov Moscow State University, 119992 Moscow, Russia.
Abstract:
A membrane-based high-throughput screening (HTS) assay for active D-amino acid oxidase (DAAO) in liquid samples as well as in intact Escherichia coli cells has been developed and optimized. The detection limit of the assay was less than 1 ng per sample. The method proposed can be used for quantitative DAAO determination in the range of 0.13 to 3.60 ng enzyme per probe. The protocol was successfully tested to screen a library of E. coli clones containing mutant DAAOs active toward target substrates.
