Tools for discovery of inhibitors of the 1-deoxy-D-xylulose 5-phosphate (DXP) synthase and DXP reductoisomerase: an approach with enzymes from the pathogenic bacterium Pseudomonas aeruginosa

Two Pseudomonas aeruginosa genes encoding the enzymes 1-deoxy-D-xylulose 5- phosphate (DXP) synthase and DXP reductoisomerase, both involved in the mev alonate-independent biosynthesis of isoprenoids, have been expressed as rec ombinant enzymes in Escherichia coli. The purified P. aeruginosa DXP reduct oisomerase was inhibited by submicromolar concentrations of the antibiotics fosmidomycin and FR-900098 in a well established method. A novel and conve nient spectrophotometric assay was developed to determine activity and inhi bition of P. aeruginosa DXP synthase. Fluoropyruvate is described as a firs t inhibitor of DXP synthase. (C) 2000 Federation of European Microbiologica l Societies. Published by Elsevier Science B.V. All rights reserved.