2020 Volume 32 Issue 187 Pages E105-E108
Endo-β-N-acetylglucosaminidases (ENGases) are glycoside hydrolases that are commonly used for glycan analysis of glycoproteins. Recently, ENGases have emerged as promising tools for glycoprotein remodeling, especially in synthesizing biotherapeutic products. Furthermore, cytosolic ENGase has been reported to cause serious symptoms of a rare genetic disorder (NGLY1-deficiency). As a tool for further study of ENGases, a pentasaccharide derivative (MM3D) was synthesized to determine the hydrolytic activity of ENGases by a fluorescence-quenching-based assay system. We succeeded in measuring the hydrolytic activities of ENGases belonging to the glycosyl hydrolase family 85, and investigated the inhibitory activities of known inhibitors. We confirmed that this assay system was suitable for high-throughput screening for inhibitors of human ENGase that have a potential to treat NGLY1-deficiency.