title: Biosynthesis of terpenoids: YgbB protein converts 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate to 2C-methyl-D-erythritol 2,4-cyclodiphosphate.
abstract: In many microorganisms, the putative orthologs of the Escherichia coli ygbB gene are tightly linked or fused to putative orthologs of ygbP, which has been shown earlier to be involved in terpenoid biosynthesis. The ygbB gene of E. coli was expressed in a recombinant E. coli strain and was shown to direct the synthesis of a soluble, 17-kDa polypeptide. The recombinant protein was found to convert 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate into 2C-methyl-D-erythritol 2,4-cyclodiphosphate and CMP. The structure of the reaction product was established by NMR spectroscopy using (13)C-labeled substrate samples. The enzyme-catalyzed reaction requires Mn(2+) or Mg(2+) but no other cofactors. Radioactivity from [2-(14)C]2C-methyl-D-erythritol 2,4-cyclodiphosphate was diverted efficiently to carotenoids by isolated chromoplasts from Capsicum annuum and, thus, was established as an intermediate in the deoxyxylulose phosphate pathway of isoprenoid biosynthesis. YgbB protein also was found to convert 4-diphosphocytidyl-2C-methyl-D-erythritol into 2C-methyl-D-erythritol 3,4-cyclophosphate. This compound does not serve as substrate for the formation of carotenoids by isolated chromoplasts and is assumed to be an in vitro product without metabolic relevance.
authors.affiliation: Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Herz
authors.forename: J
authors.initials: J
authors.lastname: Wungsintaweekul
authors.forename: C A
authors.initials: CA
authors.lastname: Schuhr
authors.forename: S
authors.initials: S
authors.lastname: Hecht
authors.forename: H
authors.initials: H
authors.lastname: Luttgen
authors.forename: S
authors.initials: S
authors.lastname: Sagner
authors.forename: M
authors.initials: M
authors.lastname: Fellermeier
authors.forename: W
authors.initials: W
authors.lastname: Eisenreich
authors.forename: M H
authors.initials: MH
authors.lastname: Zenk
authors.forename: A
authors.initials: A
authors.lastname: Bacher
authors.forename: F
authors.initials: F
authors.lastname: Rohdich
mesh_terms: D001426:Bacterial Proteins / Q000502:physiology*; D004896:Erythritol / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010710:Phosphates / Q000378:metabolism; D019761:Phosphorus-Oxygen Lyases; D018087:Plastids / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013403:Sugar Phosphates / Q000378:metabolism*
chemical_list: C406803:4-diphosphocytidyl-2C methylerythritol 2-phosphate; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D010710:Phosphates; D011994:Recombinant Proteins; D013403:Sugar Phosphates; C073443:2-methyl-butan-1,2,3,4-tetraol-2,4-cyclopyrophosphate; D019761:Phosphorus-Oxygen Lyases; C400181:ISPF protein, E coli; D004896:Erythritol