Diethylene Glycol Monoethyl Ether → Beta-D-Glucose 6-phosphate

Participants

Reaction Steps

To 0.5 ml G-6-PDH (2.5 mg/ml, 777 IV/mg) was added 20 mg G-6-PNa2, 20 mg NADH, 150 μl of carbitol and the NHS ester prepared above in 3 1 μl aliquots followed by a 0.5 μl aliquot over a period of about 3 hrs while maintaining the pH with 0.1 N NaOH between 8.55-8.77, to provide an amitriptyline-enzyme mole ratio of about 58.3. The reaction mixture was then chromatographed over Sephadex G-50 M (80 ml column) employing 0.05 M carbonate buffer (pH9.6) as eluent, collecting 20 drop fractions with fractions 16-21 pooled to provide a total volume of 8 ml. The max rate as determined by a G-6-PDH assay to be described was ΔOD/min 653.3 (% deactivation 82.8) and % inhibitability using saturating amounts of anti(amitriptyline) varied from 85.3 to 90.8 depending on the source of antibody.

Provenance & Evidence