4-METHYLUMBELLIFERYL 2-ACETAMIDO-2-DEOXY-α-D-GLUCOPYRANOSIDE, A FLUOROGENIC SUBSTRATE FOR N-ACETYL-α-D-GLUCOSAMINIDASE

Article abstract of DOI:10.1016/S0008-6215(00)84698-4

Condensation of dimeric 3,4,6-tri-O-acetyl-2-deoxy-2-nitroso-α-D-glucopyranosyl chloride with 4-methylumbelliferone gave crystalline 4-methylumbelliferyl 3,4,6-tri-O-acetyl-2-deoxy-2-oximino-α-D-arabino-hexopyranoside.Acetylation of this adduct, reduction of the resulting crude O-acetyloxime with borane in oxolane, and acetylation gave the 3,4,6-tri-O-acetyl derivative of 4-methylumbelliferyl 2-acetamido-2-deoxy-α-D-glucopyranoside (1).A new sensitive assay of N-acetyl-α-D-glucosaminidase (EC 3.2.1.50) is made possible by fluorometric measurement of 4-methylumbelliferone liberated by enzymic hydrolysis of glycoside 1.Such assays are illustrated by results obtained with enzyme preparations from pig liver and human-blood serum.