Non-mevalonate isoprenoid pathway

A process for producing an intermediate in the non-mevalonate isoprenoid pathway by

• (a) adding 2C-methyl-D-erythritol 2,4-cyclopyrophosphate to an aqueous suspension of intact or lysed plastids or bacterial cells or of a protein fraction thereof;
• (b) incubating the obtained mixture for a predetermined period of time at a predetermined temperature;
• (c) optionally disrupting the plastids or cells;
• (d) separating insoluble components and polymer components from the mixture;
• (e) subjecting the obtained aqueous solution, optionally after concentration, to a reversed phase ion pair high performance liquid chromatography;
• (f) separating a fraction containing a metabolite having a retention volume downstream from the retention volume of 2C-methyl-D-erythritol 2,4-cyclopyrophosphate and upstream from the retention volume of isopentenyl pyrophosphate;
• and
• (g) isolating the intermediate from said fraction.

A method for screening chemical libraries for the presence or absence of inhibition of the biosynthesis of isoprenoids by blocking the biosynthetic conversion of 2C-methyl-D-erythritol 2,4-cyclopyrophosphate in accordance with the following steps:

• (a) preparing an aqueous mixture comprising intact or lysed plastids or bacterial cells, 2C-methyl-D-erythritol 2,4-cyclopyrophosphate and a divalent metal salt;
• (b) reacting said mixture for a predetermined period of time at a predetermined temperature;
• (c) detecting the level of conversion of 2C-methyl-D-erythritol 2,4-cyclopyrophosphate;
• (d) repeating steps (a) to (c) in the presence of a test sample of a chemical library;
• (e) determining the presence or absence of inhibition in step (d) by ascertaining whether or not the detected level of conversion is lower in the presence of the test sample.