7-(alpha-D-Galactopyranosyloxy)-4-methyl-2H-1-benzopyran-2-one → 4-Methylumbelliferone

HRID 1707925

Reaktionsgleichung

Reaktionsgleichung von HRID 1707925
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>C([O-])(=O)CC(CC([O-])=O)(C([O-])=O)O.P([O-])([O-])([O-])=O>[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]=1

Komponenten

4

Versuchsvorschrift

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 citrate-phosphate 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 minutes. 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.

Aufarbeitung

  1. custompH 4.6 solubilized in an ultrasonic bath
  2. additionThe reaction mixtures containing
  3. custom10-50 μl of enzyme preparation or cell extracts and 150 μl substrate
  4. customThe reactions were terminated with the addition of 2.33 ml of 0.1M ethylenediamine

Quellen und Nachweise

ORD-Eintrag
ord-4d0058ee890a4800b3589efe9cd77d13
Quelldatensatz
uspto-grants-1994_10
Patentnummer
US05356804

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