Rapid and flexible synthesis of 1-deoxy-d-xylulose-5-phosphate, the substrate for 1-deoxy-d-xylulose-5-phosphate reductoisomerase

Organic & Biomolecular Chemistry Pub Date: 2003-08-20 DOI: 10.1039/B307314A

Abstract

1-Deoxy-D-xylulose-5-phosphate (DXP) is a key intermediate in the non-mevalonate pathway to terpenoids in bacteria, and it is the substrate for the enzyme 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXP-R). In order to study the mechanism of DXP-R, we required a flexible synthesis of the substrate which would allow the incorporation of isotopic labels, and the variation of the two stereocentres. Thus 1,4-dihydroxypent-2-yne was selectively reduced to give the E-olefin, and selective phosphorylation of the primary alcohol followed by oxidation of the secondary alcohol gave a substrate suitable for dihydroxylation. Dihydroxylation using stoichiometric OsO4 in the presence of chiral ligands gave protected DXP in high ee. Final hydrogenolysis gave DXP in quantitative yield and high purity. DXP-R was produced by rapid cloning of the dxr gene from Escherichia coli through controlled expression and ion exchange chromatography. The synthetic DXP was fully active in enzyme assays catalysed by recombinant DXP-R.

Graphical abstract: Rapid and flexible synthesis of 1-deoxy-d-xylulose-5-phosphate, the substrate for 1-deoxy-d-xylulose-5-phosphate reductoisomerase
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