Integrated analysis of gene expression and metabolic fluxes in PHA-producing Pseudomonas putida grown on glycerol

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dc.contributor.authorBeckers, V
dc.contributor.authorPoblete-Castro, I
dc.contributor.authorTomasch, J
dc.contributor.authorWittmann, C
dc.date.accessioned2016-06-21T19:32:32Z
dc.date.available2016-06-21T19:32:32Z
dc.date.issued2016
dc.descriptionIndexación: Web of Sciencees
dc.description.abstractBackground: Given its high surplus and low cost, glycerol has emerged as interesting carbon substrate for the synthesis of value-added chemicals. The soil bacterium Pseudomonas putida KT2440 can use glycerol to synthesize medium-chain-length poly(3-hydroxyalkanoates) (mcl-PHA), a class of biopolymers of industrial interest. Here, glycerol metabolism in P. putida KT2440 was studied on the level of gene expression (transcriptome) and metabolic fluxes (fluxome), using precisely adjusted chemostat cultures, growth kinetics and stoichiometry, to gain a systematic understanding of the underlying metabolic and regulatory network. Results: Glycerol-grown P. putida KT2440 has a maintenance energy requirement [0.039 (mmolglycerol (gCDW h)-1)] that is about sixteen times lower than that of other bacteria, such as Escherichia coli, which provides a great advantage to use this substrate commercially. The shift from carbon (glycerol) to nitrogen (ammonium) limitation drives the modulation of specific genes involved in glycerol metabolism, transport electron chain, sensors to assess the energy level of the cell, and PHA synthesis, as well as changes in flux distribution to increase the precursor availability for PHA synthesis (Entner-Doudoroff pathway and pyruvate metabolism) and to reduce respiration (glyoxylate shunt). Under PHA-producing conditions (N-limitation), a higher PHA yield was achieved at low dilution rate (29.7 wt% of CDW) as compared to a high rate (12.8 wt% of CDW). By-product formation (succinate, malate) was specifically modulated under these regimes. On top of experimental data, elementary flux mode analysis revealed the metabolic potential of P. putida KT2440 to synthesize PHA and identified metabolic engineering targets towards improved production performance on glycerol. Conclusion: This study revealed the complex interplay of gene expression levels and metabolic fluxes under PHA-and and non-PHA producing conditions using the attractive raw material glycerol as carbon substrate. This knowledge will form the basis for the development of future metabolically engineered hyper-PHA-producing strains derived from the versatile bacterium P. putida KT2440.es
dc.description.urihttp://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-016-0470-2
dc.identifier.citationMicrobial Cell Factories 2016 15:73es
dc.identifier.issn1475-2859
dc.identifier.otherDOI: 10.1186/s12934-016-0470-2
dc.identifier.urihttp://repositorio.unab.cl/xmlui/handle/ria/832
dc.language.isoenes
dc.publisherBIOMED CENTRALes
dc.subjectPseudomonas putida KT2440es
dc.subjectGlycerol metabolismes
dc.subjectTranscriptomees
dc.subjectMetabolic flux analysises
dc.subjectFlux balance analysises
dc.subjectElementary flux modeses
dc.subjectPolyhydroxyalkanoateses
dc.subjectNitrogen and carbon limitationes
dc.subjectCHAIN-LENGTH POLYHYDROXYALKANOATEes
dc.subjectESCHERICHIA-COLIes
dc.subjectCONTINUOUS CULTURESes
dc.subjectBACILLUS-SUBTILISes
dc.subjectBACTERIAL POLYHYDROXYALKANOATESes
dc.subjectCORYNEBACTERIUM-GLUTAMICUMes
dc.subjectIN-VIVOes
dc.subjectKT2440es
dc.subjectBIOSYNTHESISes
dc.subjectPATHWAYes
dc.titleIntegrated analysis of gene expression and metabolic fluxes in PHA-producing Pseudomonas putida grown on glyceroles
dc.typeArtículoes
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