Regeneration of NAD+ cofactor by photosensitized electron transfer in an immobilized alcohol dehydrogenase system

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Abstract
The irradiation with visible light of a photosensitizer dye like methylene blue was used to regenerate by electron transfer the oxidized form of a pyridine nucleotide coenzyme (NAD+). The process has been studied on a common enzymatic reaction: ethanol oxidation by alcohol–NAD+ oxidoreductase immobilized on polyacrylamide gel or porous glass balls. In the experimental conditions used, the initial NAD+ recycling rates were 2.33 × 104 cycles/h (polyacrylamide) and 3 × 104 cycles/h (glass balls). A total number of 49.5 × 104 cycles was obtained for 13 runs of 2 h. The enzyme immobilization strongly increased its stability: after 28 days at 20°C, the residual activity was 25% of the initial value.