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Investigation of the methylerythritol 4-phosphate pathway for microbial terpenoid production through metabolic control analysis

dc.contributor.authorVolke, Daniel C.en_ZA
dc.contributor.authorRohwer, Johannen_ZA
dc.contributor.authorFischer, Raineren_ZA
dc.contributor.authorJennewein, Stefanen_ZA
dc.date.accessioned2019-11-11T06:31:51Z
dc.date.available2019-11-11T06:31:51Z
dc.date.issued2019-11-05
dc.identifier.citationVolke, D. C., et al. 2019. Investigation of the methylerythritol 4-phosphate pathway for microbial terpenoid production through metabolic control analysis. Microbial Cell Factories, 18:192, doi:10.1186/s12934-019-1235-5
dc.identifier.issn1475-2859 (online)
dc.identifier.otherdoi:10.1186/s12934-019-1235-5
dc.identifier.urihttp://hdl.handle.net/10019.1/106819
dc.descriptionCITATION: Volke, D. C., et al. 2019. Investigation of the methylerythritol 4-phosphate pathway for microbial terpenoid production through metabolic control analysis. Microbial Cell Factories, 18:192, doi:10.1186/s12934-019-1235-5.
dc.descriptionThe original publication is available at https://microbialcellfactories.biomedcentral.com
dc.description.abstractBackground: Terpenoids are of high interest as chemical building blocks and pharmaceuticals. In microbes, terpenoids can be synthesized via the methylerythritol phosphate (MEP) or mevalonate (MVA) pathways. Although the MEP pathway has a higher theoretical yield, metabolic engineering has met with little success because the regulation of the pathway is poorly understood. Results: We applied metabolic control analysis to the MEP pathway in Escherichia coli expressing a heterologous isoprene synthase gene (ispS). The expression of ispS led to the accumulation of isopentenyl pyrophosphate (IPP)/ dimethylallyl pyrophosphate (DMAPP) and severely impaired bacterial growth, but the coexpression of ispS and isopentenyl diphosphate isomerase (idi) restored normal growth and wild-type IPP/DMAPP levels. Targeted proteomics and metabolomics analysis provided a quantitative description of the pathway, which was perturbed by randomizing the ribosome binding site in the gene encoding 1-deoxyxylulose 5-phosphate synthase (Dxs). Dxs has a flux control coefficient of 0.35 (i.e., a 1% increase in Dxs activity resulted in a 0.35% increase in pathway flux) in the isoprene-producing strain and therefore exerted significant control over the flux though the MEP pathway. At higher dxs expression levels, the intracellular concentration of 2-C-methyl-d-erythritol-2,4-cyclopyrophosphate (MEcPP) increased substantially in contrast to the other MEP pathway intermediates, which were linearly dependent on the abundance of Dxs. This indicates that 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase (IspG), which consumes MEcPP, became saturated and therefore limited the flux towards isoprene. The higher intracellular concentrations of MEcPP led to the efflux of this intermediate into the growth medium. Discussion: These findings show the importance of Dxs, Idi and IspG and metabolite export for metabolic engineering of the MEP pathway and will facilitate further approaches for the microbial production of valuable isoprenoids.
dc.description.urihttps://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-019-1235-5
dc.format.extent15 pagesen_ZA
dc.language.isoen_ZAen_ZA
dc.publisherBMC (part of Springer Nature)en_ZA
dc.subjectMetabolic control analysisen_ZA
dc.subjectTerpenoidsen_ZA
dc.subjectMicrobiologyen_ZA
dc.subjectIsopreneen_ZA
dc.titleInvestigation of the methylerythritol 4-phosphate pathway for microbial terpenoid production through metabolic control analysisen_ZA
dc.typeArticleen_ZA
dc.date.updated2019-11-10T04:20:16Z
dc.description.versionPublisher's version
dc.rights.holderAuthors retain copyrighten_ZA


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