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Kinetic modeling of plasmid bioproduction in Escherichia coli DH5α cultures over different carbon-source compositions

2014-09-30Journal article Restricted access
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dc.contributor.authorLopes, Marta B.
dc.contributor.authorMartins, Gabriel
dc.contributor.authorCalado, Cecília
dc.date.accessioned2020-09-16T13:55:56Z
dc.date.available2020-09-16T13:55:56Z
dc.date.issued2014-09-30
dc.description.abstractThe need for the development of economic high plasmid production in Escherichia coli cultures is emerging, as a result of the latest advances in DNA vaccination and gene therapy. In order to contribute to achieve that, a model describing the kinetics involved in the bioproduction of plasmid by recombinant E. coli DH5α is presented, as an attempt to understand the complex and non-linear metabolic relationships and the plasmid production occurring in dynamic batch culture environments, run under different media compositions of glucose and glycerol, that result in distinct maximum biomass growths (between 8.2 and 12.8 g DCW/L) and specific plasmid productions (between 1.1 and 7.4 mg/g DCW). The model based on mass balance equations for biomass, glucose, glycerol, acetate and plasmid accurately described different culture behaviors, using either glucose or glycerol as carbon source, or mixtures of both. From the 17 parameters obtained after model simplification, the following 10 parameters were found to be independent of the carbon source composition: the substrate affinity constants, the inhibitory constants of biomass growth on glycerol by glucose, of biomass growth on acetate by glycerol and the global biomass growth by acetate, and the yields of biomass on acetate, acetate on glucose and glycerol, and plasmid on glucose. The parameters that depend on the culture composition, and that might explain the differences found between cultures, were: maximum specific growth rates on glucose, glycerol and acetate; biomass yield on glucose and glycerol; and plasmid yield on glycerol and acetate. Moreover, a crucial role of acetate in the plasmid production was revealed by the model, with most of plasmid production being associated to the acetate consumption. The model provides meaningful insight on the E. coli dynamic cell behavior concerning the plasmid bioproduction, which might lead to important guidelines for culture optimization and process scale-up and control.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationLOPES, Marta B.; MARTINS, Gabriel; CALADO, Cecília R. C. – Kinetic modeling of plasmid bioproduction in Escherichia coli DH5α cultures over different carbon-source compositions. Journal of Biotechnology. ISSN 0168-1656. Vol. 186 (2014), pp. 38-48pt_PT
dc.identifier.doi10.1016/j.jbiotec.2014.06.022pt_PT
dc.identifier.issn0168-1656
dc.identifier.urihttp://hdl.handle.net/10400.21/12239
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://reader.elsevier.com/reader/sd/pii/S0168165614003137?token=0D91FD807985E585F6C8FE81727EF7326945F79792CD5AB57B1E43C8AECE79B0E818CDF9E55628E6CF598C7BD1FE7F92pt_PT
dc.subjectEscherichia colipt_PT
dc.subjectKinetic modelpt_PT
dc.subjectMultiple substratept_PT
dc.subjectPlasmidpt_PT
dc.subjectAcetatept_PT
dc.subjectFermentationpt_PT
dc.titleKinetic modeling of plasmid bioproduction in Escherichia coli DH5α cultures over different carbon-source compositionspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBIO%2F69242%2F2006/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F73758%2F2010/PT
oaire.citation.endPage48pt_PT
oaire.citation.startPage38pt_PT
oaire.citation.titleJournal of Biotechnologypt_PT
oaire.citation.volume186pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStreamSFRH
person.familyNameB. Lopes
person.familyNameCalado
person.givenNameMarta
person.givenNameCecília
person.identifier1195979
person.identifier130332
person.identifier.ciencia-idFD16-A07F-7B12
person.identifier.ciencia-id9418-E320-3177
person.identifier.orcid0000-0002-4135-1857
person.identifier.orcid0000-0002-5264-9755
person.identifier.ridF-5378-2011
person.identifier.ridE-2102-2014
person.identifier.scopus-author-id55489480400
person.identifier.scopus-author-id6603163260
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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