Use of a polymer-bound flavin derivative for the rapid regeneration of NAD(P)+ from NAD(P)H in dehydrogenase systems.
An aldehyde derivative of riboflavin was covalently attached by reductive alkylation to soluble polycationic supports. The flavopolymers so obtained were stable under operational conditions. The catalytic efficiency towards oxidation of NADH by these flavopolymers was demonstrated, and the kinetic parameters (Km and kcat) revealed an overall catalytic efficiency (kcat/Km) 185-fold greater compared to riboflavin. Various factors affecting the chemical regeneration of NAD+ from NADH such as pH, ionic strength, nature of the buffer etc. were studied. The most interesting result was the highly favourable influence of borate ions which increased the reaction rate by a factor 2-4 compared to the other buffers. The flavopolymers are very effective for in situ recycling of NAD(P)+. With up to 300-fold NADH----NAD+ conversions for the system using yeast alcohol dehydrogenase and up to 1500-fold NADPH----NADP+ regenerations for the system using glucose-6-phosphate dehydrogenase. These flavopolymers are superior to previous chemical recycling systems.
D000426:Alcohol Dehydrogenase; D002021:Buffers; D005415:Flavins; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D009243:NAD; D009249:NADP; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D010088:Oxidoreductases; D011108:Polymers; D012256:Riboflavin / Q000031:analogs & derivatives
D002021:Buffers; D005415:Flavins; D011108:Polymers; D009243:NAD; D009249:NADP; D010088:Oxidoreductases; D000426:Alcohol Dehydrogenase; D012256:Riboflavin