Electrochemical bioreactor with immobilized glucose-6-phosphate dehydrogenase on the rotating graphite disc electrode modified with phenazine methosulfate.

Physical adsorption, covalent binding through the carbodiimide reaction between the surface carboxyl group and the amino group of the protein, and the crosslinking method with bovine serum albumin by glutaraldehyde were applied for the immobilization of glucose-6-phosphate dehydrogenase (G6PDH) on a graphite electrode. Among those, the crosslinking method was employed for its highest apparent enzyme activity per unit surface area. Phenazine methosulfate (PMS), as a mediator for the electrochemical oxidation of NADH, was also immobilized on the graphite surface through adsorption. The conjugation reaction of G6PDH and the electrochemical oxidation of NADH were confirmed by cyclic voltammetry and the constant potential electrochemical reaction. An electrochemical bioreactor system was established by using a rotating disc graphite electrode with G6PDH immobilized. The coenzyme, NAD, was effectively recycled between the electrochemical and the enzymatic reactions.