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Metabolic proļ¬ling of recombinant Escherichia coli cultivations based on high-throughput FT-MIR spectroscopic analysis

2017Journal article Restricted access
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dc.contributor.authorSales, Kevin C.
dc.contributor.authorRosa, Filipa
dc.contributor.authorRibeiro Da Cunha, Bernardo
dc.contributor.authorSampaio, Pedro
dc.contributor.authorB. Lopes, Marta
dc.contributor.authorCalado, CecĆ­lia
dc.date.accessioned2017-11-07T10:56:04Z
dc.date.available2017-11-07T10:56:04Z
dc.date.issued2017
dc.description.abstractEscherichia coli is one of the most used host microorganism for the production of recombinant products, such as heterologous proteins and plasmids. However, genetic, physiological and environmental factors inļ¬‚uence the plasmid replication and cloned gene expression in a highly complex way. To control and optimize the recombinant expression system performance, it is very important to understand this complexity. Therefore, the development of rap-id, highly sensitive and economic analytical methodologies, which enable the simultaneous characterization of the heterologous product synthesis and physiologic cell behavior under a variety of culture conditions, is highly desirable. For that, the metabolic proļ¬le of recombinant E. coli cultures producing the pVAX-lacZ plasmid model was analyzed by rapid, economic and high-throughput Fourier Transform Mid-Infrared (FT-MIR) spectroscopy. The main goal of the present work is to show as the simultaneous multivariate data analysis by principal component analysis (PCA) and direct spectral analysis could represent a very interesting tool to monitor E. coli culture processes and acquire relevant information according to current quality regulatory guidelines. While PCA allowed capturing the energetic metabolic state of the cell, e.g. by identifying different C-sources consumption phases, direct FT-MIR spectral analysis allowed obtaining valuable biochemical and metabolic information along the cell culture, e.g. lipids, RNA, protein synthesis and turnover metabolism. The information achieved by spectral multivariate data and direct spectral analyses complement each other and may contribute to understand the complex interrelationships between the recombinant cell metabolism and the bioprocess environment towards more economic and robust processes design according to Quality by Design framework.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationSALES, Kevin C.; [at al] - Metabolic proļ¬ling of recombinant Escherichia coli cultivations based on high-throughput FT-MIR spectroscopic analysis. Biotechnology Progress. ISSN 8756-7938. Vol. 33, N.Āŗ 2 (2017), pp 285-298pt_PT
dc.identifier.doi10.1002/btpr.2378pt_PT
dc.identifier.issn8756-7938
dc.identifier.issn1520-6033
dc.identifier.urihttp://hdl.handle.net/10400.21/7460
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherAmerican Institute of Chemical Engineerspt_PT
dc.relation.publisherversionhttp://onlinelibrary.wiley.com/doi/10.1002/btpr.2378/epdfpt_PT
dc.subjectBioprocesspt_PT
dc.subjectFT-MIR spectroscopypt_PT
dc.subjectMetabolic proļ¬lingpt_PT
dc.subjectPCApt_PT
dc.titleMetabolic proļ¬ling of recombinant Escherichia coli cultivations based on high-throughput FT-MIR spectroscopic analysispt_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.endPage298pt_PT
oaire.citation.issue2pt_PT
oaire.citation.startPage285pt_PT
oaire.citation.titleBiotechnology Progresspt_PT
oaire.citation.volume33pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStreamSFRH
person.familyNameRibeiro da Cunha
person.familyNameSampaio
person.familyNameB. Lopes
person.familyNameCalado
person.givenNameBernardo
person.givenNamePedro
person.givenNameMarta
person.givenNameCecĆ­lia
person.identifier1251979
person.identifier1195979
person.identifier130332
person.identifier.ciencia-idEA1E-4BEA-A01E
person.identifier.ciencia-idAF12-6ABA-43D8
person.identifier.ciencia-idFD16-A07F-7B12
person.identifier.ciencia-id9418-E320-3177
person.identifier.orcid0000-0002-0303-9416
person.identifier.orcid0000-0003-2917-4904
person.identifier.orcid0000-0002-4135-1857
person.identifier.orcid0000-0002-5264-9755
person.identifier.ridP-6154-2017
person.identifier.ridF-5378-2011
person.identifier.ridE-2102-2014
person.identifier.scopus-author-id57211629814
person.identifier.scopus-author-id24178064100
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.nameFundaĆ§Ć£o para a CiĆŖncia e a Tecnologia
project.funder.nameFundaĆ§Ć£o para a CiĆŖncia e a Tecnologia
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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