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Escherichia coli engineered to synthesize isopentenyl diphosphate and dimethylallyl diphosphate from mevalonate: a novel system for the genetic analysis of the 2-C-methyl-d-erythritol 4-phosphate pathway for isoprenoid biosynthesis.
Biochem J. 2001 Jan 01; 353(Pt 1):59-67.BJ

Abstract

Isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP) constitute the basic building block of isoprenoids, a family of compounds that is extraordinarily diverse in structure and function. IPP and DMAPP can be synthesized by two independent pathways: the mevalonate pathway and the recently discovered 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway. Although the MEP pathway is essential in most eubacteria, algae and plants and has enormous biotechnological interest, only some of its steps have been determined. We devised a system suitable for the genetic analysis of the MEP pathway in Escherichia coli. A synthetic operon coding for yeast 5-diphosphomevalonate decarboxylase, human 5-phosphomevalonate kinase, yeast mevalonate kinase and E. coli isopentenyl diphosphate isomerase was incorporated in the chromosome of this bacterium. The expression of this operon allowed the synthesis of IPP and DMAPP from mevalonate added exogenously and complementation of lethal mutants of the MEP pathway. We used this system to show that the ygbP, ychB and ygbB genes are essential in E. coli and that the steps catalysed by the products of these genes belong to the trunk line of the MEP pathway.

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Authors+Show Affiliations

Campos N
Department de Bioquímica i Biologia Molecular, Facultat de Química, Universitat de Barcelona, C/Martí i Franquès 1, 08028 Barcelona, Spain. campos@sun.bq.ub.es
Rodríguez-Concepción M
No affiliation info available
Sauret-Güeto S
No affiliation info available
Gallego F
No affiliation info available
Lois LM
No affiliation info available
Boronat A
No affiliation info available

MeSH

Bacterial ProteinsCarbon-Carbon Double Bond IsomerasesCarboxy-LyasesChromosomes, BacterialDNA, RecombinantErythritolEscherichia coliEscherichia coli ProteinsGene DeletionGenes, BacterialGenes, EssentialGenes, FungalGenes, LethalGenetic EngineeringHemiterpenesHumansMevalonic AcidNucleotidyltransferasesOperonOrganophosphorus CompoundsPhenotypePhosphorus-Oxygen LyasesPhosphotransferases (Alcohol Group Acceptor)Phosphotransferases (Phosphate Group Acceptor)Polyisoprenyl PhosphatesRecombination, GeneticSugar Phosphates

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11115399

Citation

Campos, N, et al. "Escherichia Coli Engineered to Synthesize Isopentenyl Diphosphate and Dimethylallyl Diphosphate From Mevalonate: a Novel System for the Genetic Analysis of the 2-C-methyl-d-erythritol 4-phosphate Pathway for Isoprenoid Biosynthesis." The Biochemical Journal, vol. 353, no. Pt 1, 2001, pp. 59-67.
Campos N, Rodríguez-Concepción M, Sauret-Güeto S, et al. Escherichia coli engineered to synthesize isopentenyl diphosphate and dimethylallyl diphosphate from mevalonate: a novel system for the genetic analysis of the 2-C-methyl-d-erythritol 4-phosphate pathway for isoprenoid biosynthesis. Biochem J. 2001;353(Pt 1):59-67.
Campos, N., Rodríguez-Concepción, M., Sauret-Güeto, S., Gallego, F., Lois, L. M., & Boronat, A. (2001). Escherichia coli engineered to synthesize isopentenyl diphosphate and dimethylallyl diphosphate from mevalonate: a novel system for the genetic analysis of the 2-C-methyl-d-erythritol 4-phosphate pathway for isoprenoid biosynthesis. The Biochemical Journal, 353(Pt 1), 59-67.
Campos N, et al. Escherichia Coli Engineered to Synthesize Isopentenyl Diphosphate and Dimethylallyl Diphosphate From Mevalonate: a Novel System for the Genetic Analysis of the 2-C-methyl-d-erythritol 4-phosphate Pathway for Isoprenoid Biosynthesis. Biochem J. 2001 Jan 1;353(Pt 1):59-67. PubMed PMID: 11115399.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Escherichia coli engineered to synthesize isopentenyl diphosphate and dimethylallyl diphosphate from mevalonate: a novel system for the genetic analysis of the 2-C-methyl-d-erythritol 4-phosphate pathway for isoprenoid biosynthesis. AU - Campos,N, AU - Rodríguez-Concepción,M, AU - Sauret-Güeto,S, AU - Gallego,F, AU - Lois,L M, AU - Boronat,A, PY - 2000/12/15/pubmed PY - 2001/3/3/medline PY - 2000/12/15/entrez SP - 59 EP - 67 JF - The Biochemical journal JO - Biochem J VL - 353 IS - Pt 1 N2 - Isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP) constitute the basic building block of isoprenoids, a family of compounds that is extraordinarily diverse in structure and function. IPP and DMAPP can be synthesized by two independent pathways: the mevalonate pathway and the recently discovered 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway. Although the MEP pathway is essential in most eubacteria, algae and plants and has enormous biotechnological interest, only some of its steps have been determined. We devised a system suitable for the genetic analysis of the MEP pathway in Escherichia coli. A synthetic operon coding for yeast 5-diphosphomevalonate decarboxylase, human 5-phosphomevalonate kinase, yeast mevalonate kinase and E. coli isopentenyl diphosphate isomerase was incorporated in the chromosome of this bacterium. The expression of this operon allowed the synthesis of IPP and DMAPP from mevalonate added exogenously and complementation of lethal mutants of the MEP pathway. We used this system to show that the ygbP, ychB and ygbB genes are essential in E. coli and that the steps catalysed by the products of these genes belong to the trunk line of the MEP pathway. SN - 0264-6021 UR - https://www.unboundmedicine.com/medline/citation/11115399/Escherichia_coli_engineered_to_synthesize_isopentenyl_diphosphate_and_dimethylallyl_diphosphate_from_mevalonate:_a_novel_system_for_the_genetic_analysis_of_the_2_C_methyl_d_erythritol_4_phosphate_pathway_for_isoprenoid_biosynthesis_ DB - PRIME DP - Unbound Medicine ER -