Specific enzyme inhibitors in vitamin biosynthesis. Part 5. Purification of riboflavin synthase by affinity chromatography using 7-oxolumazines

Abstract

6-Carboxyethyl-7-oxo-8-D-ribityl-lumazine (3) has been designed based on information about the mechanism of the enzymic reaction catalysed by riboflayin synthase (E.C. 2.5.1.9). This compound has been synthesised and used as a specific ligand for the affinity chromatography purification of the enzyme by attaching it in three different ways to Sepharose 4B. The effectiveness of each of these polymeric supports has been compared in order to define an optimised method for the purification of the enzyme.