Assessment of enzymatic activity of G6PDH in artificial conditions

Known amounts (0.05, 0.1, 0.25, 0.5, 1 mU) of glucose-6-phosphate dehydrogenase (G6PDH) were added to a 0.2 ml reaction solution containing 50 mM Tris-HCl buffer (pH 7.5), 5 mM glucose-6-phosphate (G6P), 2 mM NADP⁺, 50 mM MgCl₂, and 10 μM mBFP. One unit of G6PDH activity is defined as the amount of enzyme required for the formation of 1 μmol NADPH per min at pH 7.5 and 30 °C. The temperatures of the fluorometer and microplate were maintained at 30 °C. Theoretically, 1 mU of G6PDH can produce 40 nmol of NADPH from 400 nmol of NADP⁺.