7-(alpha-D-Galactopyranosyloxy)-4-methyl-2H-1-benzopyran-2-one → 4-Methylumbelliferone
반응식
O[C@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O.[CH3:13][C:14]1[C:20]2[CH:21]=[CH:22][C:23]([O:25]C3OC(CO)C(O)C(O)C3O)=[CH:24][C:19]=2[O:18][C:16](=[O:17])[CH:15]=1>>[CH3:13][C:14]1[C:20]2[CH:21]=[CH:22][C:23]([OH:25])=[CH:24][C:19]=2[O:18][C:16](=[O:17])[CH:15]=1참가 물질
3
실험 절차
α-Gal A was assayed using 4-methylumbelliferyl-α-D-galactopyranoside (4-MU-α-Gal) as previously described (Bishop, et al., 1980, In Enzyme Therapy in Genetic Diseases:2. Desnick, R. J. (Ed.). Alan R. Liss, Inc. New York, p. 17). Briefly, a stock solution of 5 mM 4-MU was prepared in 0.1M citratephosphate buffer, pH 4.6 solubilized in an ultrasonic bath. The reaction mixtures containing 10-50 μl of enzyme preparation or cell extracts and 150 μl substrate, were incubated at 37° C. for 10-30 min. The reactions were terminated with the addition of 2.33 ml of 0.1M ethylenediamine. One unit of activity is that amount of enzyme which hydrolyzes 1 nmol of substrate/hour.
후처리
- custompH 4.6 solubilized in an ultrasonic bath
- addition