A fluorometric assay of ceramide glycanase with 4-methylumbelliferyl beta-D-lactoside derivatives
Wang, L.X.; Pavlova, N.V.; Li, S.C.; Li, Y.T.; Lee, Y.C.
Glycoconjugate Journal 13(3): 359-365
4-Methylumbelliferyl 6'-O-benzyl-beta-D-lactoside (6'Bn-MU-Lac) and some related compounds were synthesized via different selective reactions including phase-transfer glycosylation. Their suitability as substrates for a fluorometric assay of ceramide glycanase (CGase) was evaluated. Among others, the 6'Bn-MU-Lac, which is resistant to exogalactosidase, was found to be a suitable substrate for routine assay of the CGase activity. For American leech CGase, the K(m) value is 0.232 mM at pH 5.