A closed loop system for the synthesis of reduced nicotinamide adenine dinucleotide (NADH) from oxidized NAD is described using isolated and purified Alcohol dehydroganese (ADH) from baker’s yeast, immobilized on phenolic resin by covalent attachment. The NADH formed is removedby in-line diethylaminoethyl cellulose (DEAE-cellulose) ion exchange column and the progress of the reaction is monitored by high performance liquid chromatography (HPLC). The effect of regent concentrations and key physical parameters are optimized for the synthesis. Calibration datais linear over the range of 0.2-1.0 mM NAD with relative standard deviation (n=5) in the range 0.6-1.9%. the ADH immobilized onto phenolic resin was stable for 3 months at 4oC and lost its activity of 4.0±1.2% at room temperature during synthesis.
L RISHI ,M MASOOM YASINZIA ,A NABI ,M YAQOOB ,
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