Regeneration of NADPH by Cactus Chloroplasts : Coupling Reaction with P450 Monooxygenase

概要

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A stable photoreaction system was developed using cactus chloroplasts coupled with the fused enzyme between rat cytochrome P4501A1 (CYP1A1) and yeast NADPH-cytochrome P450 oxidoreductase expressed in yeast mirosomes. The fused enzyme catalyzed o-deethylation of 7-ethoxycoumarin to 7-hydroxycoumarin using molecular oxygen and NADPH produced by phososystems I and II. Cactus chloroplast were found to constitute a good material for use as a NADPH-regenerating system in bioreactors. For NADPH formation, cactus chloroplasts showed twice as much activity as those of spinach when the same chlorophill concentration was used at 30℃, as well as being more thermostable. The optimal temperature for NADPH formation with cactus chloroplasts was 42.5℃, whereas for spinach it ranged from 20 to 30℃. Cactus chloroplasts can thus be applied in bioreactor NADPH-regenerating systems using oxido-reductive enzymes like P450s.
社団法人日本生物工学会の論文
1997-10-25