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dc.contributor.authorLien, Stina Katrine
dc.contributor.authorNiedenführ, Sebastian
dc.contributor.authorSletta, Håvard
dc.contributor.authorNöh, Katharina
dc.contributor.authorBruheim, Per
dc.date.accessioned2015-05-07T06:53:51Z
dc.date.accessioned2015-05-21T08:48:21Z
dc.date.available2015-05-07T06:53:51Z
dc.date.available2015-05-21T08:48:21Z
dc.date.issued2015
dc.identifier.citationBMC Systems Biology 2015, 9(6)nb_NO
dc.identifier.issn1752-0509
dc.identifier.urihttp://hdl.handle.net/11250/283846
dc.description.abstractBackground: The bacterium Pseudomonas fluorescens switches to an alginate-producing phenotype when the pleiotropic anti-sigma factor MucA is inactivated. The inactivation is accompanied by an increased biomass yield on carbon sources when grown under nitrogen-limited chemostat conditions. A previous metabolome study showed significant changes in the intracellular metabolite concentrations, especially of the nucleotides, in mucA deletion mutants compared to the wild-type. In this study, the P. fluorescens SBW25 wild-type and an alginate non-producing mucA- ΔalgC double-knockout mutant are investigated through model-based 13C-metabolic flux analysis (13C-MFA) to explore the physiological consequences of MucA inactivation at the metabolic flux level. Intracellular metabolite extracts from three carbon labelling experiments using fructose as the sole carbon source are analysed for 13C-label incorporation in primary metabolites by gas and liquid chromatography tandem mass spectrometry. R es u l t s : From mass isotopomer distribution datasets, absolute intracellular metabolic reaction rates for the wild type and the mutant are determined, revealing extensive reorganisation of carbon flux through central metabolic pathways in response to MucA inactivation. The carbon flux through the Entner-Doudoroff pathway was reduced in the mucA- ΔalgC mutant, while flux through the pentose phosphate pathway was increased. Our findings also indicated flexibility of the anaplerotic reactions through down-regulation of the pyruvate shunt in the mucA- ΔalgC mutant and up-regulation of the glyoxylate shunt. Conclusions: Absolute metabolic fluxes and metabolite levels give detailed, integrated insight into the physiology of this industrially, medically and agriculturally important bacterial species and suggest that the most efficient way of using a mucA- mutant as a cell factory for alginate production would be to use non-growing conditions and nitrogen deprivation. Keywords: Pseudomonas fluorescens, Anti-sigma factor MucA, Fluxome and fluxomics, Carbon labelling experiments, GC-MS/MS, LC-MS/MS, 13C-metabolic flux analysisnb_NO
dc.language.isoengnb_NO
dc.publisherBioMed Centralnb_NO
dc.titleFluxome study of Pseudomonas fluorescens reveals major reorganisation of carbon flux through central metabolic pathways in response to inactivation of the anti-sigma factor MucAnb_NO
dc.typeJournal articlenb_NO
dc.typePeer revieweden_GB
dc.date.updated2015-05-07T06:53:51Z
dc.source.volume9nb_NO
dc.source.journalBMC Systems Biologynb_NO
dc.source.issue6nb_NO
dc.identifier.doi10.1186/s12918-015-0148-0
dc.identifier.cristin1234847
dc.description.localcode© 2015 Lien et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article unless otherwise stated.nb_NO


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