High-Performance Liquid Chromatography of Derivatives Involved in the Terminal Steps of Tetracycline Biosynthesis inStreptomyces Aureofaciens

摘要

A simple and reproducible method was developed for the analysis of tetracycline derivatives involved in the last two steps of tetracycline biosynthetic pathway inStreptomyces aureofaciens. The method is based on a gradient liquid chromatographic separation of the compounds using a microbore octadecyl silica column. Beside separation of a mixture of standards, the method was used for separation, detection and quantitation of dehydrotetracycline and tetracycline prepared enzymaticallyin vitrousing anhydrotetracycline oxygenase and tetracycline dehydrogenase, respectively, isolated fromS. aureofaciens. The method permits a simple and accurate characterization of kinetics of corresponding enzymatic activities.