title: Biosynthesis of terpenoids: 1-deoxy-D-xylulose-5-phosphate reductoisomerase from Escherichia coli is a class B dehydrogenase.
abstract: 1-Deoxy-D-xylulose-5-phosphate is converted into 2-C-methyl-D-erythritol-4-phosphate by the catalytic action of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (Dxr protein) using NADPH as cofactor. The stereochemical features of this reaction were investigated in in vitro experiments with the recombinant Dxr protein of Escherichia coli using (4R)- or (4S)-[4-(2)H(1)]NADPH as coenzyme. The enzymatically formed 2-C-methyl-D-erythritol-4-phosphate was isolated and converted into 1,2:3,4-di-O-isopropylidene-2-C-methyl-D-erythritol; NMR spectroscopic investigation of this derivative indicated that only (4S)-[4-(2)H(1)]NADPH affords 2-C-methyl-D-erythritol-4-phosphate labelled exclusively in the H(Re) position of C-1. Stereospecific transfer of H(Si) from C-4 of the cofactor identifies the Dxr protein of E. coli as a class B dehydrogenase.
authors.affiliation: Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstr. 4, D-85747, Garching, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Radykewicz
authors.forename: F
authors.initials: F
authors.lastname: Rohdich
authors.forename: J
authors.initials: J
authors.lastname: Wungsintaweekul
authors.forename: S
authors.initials: S
authors.lastname: Herz
authors.forename: K
authors.initials: K
authors.lastname: Kis
authors.forename: W
authors.initials: W
authors.lastname: Eisenreich
authors.forename: A
authors.initials: A
authors.lastname: Bacher
authors.forename: M H
authors.initials: MH
authors.lastname: Zenk
authors.forename: D
authors.initials: D
authors.lastname: Arigoni
mesh_terms: D019747:Aldose-Ketose Isomerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D009682:Magnetic Resonance Spectroscopy; D009097:Multienzyme Complexes / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D013729:Terpenes / Q000378:metabolism*
chemical_list: D009097:Multienzyme Complexes; D013729:Terpenes; D010088:Oxidoreductases; C114231:1-deoxy-D-xylulose 5-phosphate reductoisomerase; D019747:Aldose-Ketose Isomerases