Abstract
A simple biosynthetic method for the preparation of radioactive flavin coenzymes from [2- 14 C]riboflavin has been developed. The anaerobic bacterium Clostridium kluyveri during growth perferentially utilizes added riboflavin for FMN and FAD synthesis. About 80% of the radioactivity is recovered as flavin nucleotides when 1 μ m of labeled riboflavin is added to the medium. The total flavin content is about 1.7 μ mol/g wet cells of which 75% is FAD and 25% FMN. FMN and FAD are separated by DEAE-cellulose chromatography. Salt and acid are removed by passing through Florisil, and the flavin nucleotides are eluted with 5% pyridine. By this method radiochemically pure FAD and FMN are obtained. From the radioactivity added as [2- 14 C]riboflavin 50% is recovered as FAD and 25% as FMN.
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