Biosynthesis of Terpenoids: Efficient Multistep Biotransformation Procedures Affording Isotope-Labeled 2<i>C</i>-Methyl-<scp>d</scp>-erythritol 4-Phosphate Using Recombinant 2<i>C</i>-Methyl-<scp>d</scp>-erythritol 4-Phosphate Synthase

Abstract

This paper describes the recombinant expression of the ispC gene of Escherichia coli specifying 2C-methyl-D-erythritol 4-phosphate synthase in a modified form that can be purified efficiently by metal-chelating chromatography. The enzyme was used for the preparation of isotope-labeled 2C-methyl-D-erythritol 4-phosphate employing isotope-labeled glucose and pyruvate as starting materials. The simple one-pot methods described afford numerous isotopomers of 2C-methyl-D-erythritol 4-phosphate carrying (3)H, (13)C, or (14)C from commercially available precursors. The overall yield based on the respective isotope-labeled starting material is approximately 50%.