Increasing the Pentose Phosphate Pathway Flux to Improve Plasmid DNA Production in Engineered E. coli

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dc.authoridLara, Alvaro/0000-0003-3535-7619|Kunert, Flavio/0009-0008-9297-1854
dc.authorwosidLara, Alvaro/A-8397-2012
dc.contributor.authorde la Cruz, Mitzi
dc.contributor.authorKunert, Flavio
dc.contributor.authorTaymaz-Nikerel, Hilal
dc.contributor.authorSigala, Juan-Carlos
dc.contributor.authorGosset, Guillermo
dc.contributor.authorBuchs, Jochen
dc.contributor.authorLara, Alvaro R.
dc.date.accessioned2024-07-18T20:50:42Z
dc.date.available2024-07-18T20:50:42Z
dc.date.issued2024
dc.departmentİstanbul Bilgi Üniversitesien_US
dc.description.abstractThe demand of plasmid DNA (pDNA) as a key element for gene therapy products, as well as mRNA and DNA vaccines, is increasing together with the need for more efficient production processes. An engineered E. coli strain lacking the phosphotransferase system and the pyruvate kinase A gene has been shown to produce more pDNA than its parental strain. With the aim of improving pDNA production in the engineered strain, several strategies to increase the flux to the pentose phosphate pathway (PPP) were evaluated. The simultaneous consumption of glucose and glycerol was a simple way to increase the growth rate, pDNA production rate, and supercoiled fraction (SCF). The overexpression of key genes from the PPP also improved pDNA production in glucose, but not in mixtures of glucose and glycerol. Particularly, the gene coding for the glucose 6-phosphate dehydrogenase (G6PDH) strongly improved the SCF, growth rate, and pDNA production rate. A linear relationship between the G6PDH activity and pDNA yield was found. A higher flux through the PPP was confirmed by flux balance analysis, which also estimates relevant differences in fluxes of the tricarboxylic acid cycle. These results are useful for developing further cell engineering strategies to increase pDNA production and quality.en_US
dc.description.sponsorshipCONAHCyT [A1-S-8646]; MDPIen_US
dc.description.sponsorshipThis work was supported by CONAHCyT grant A1-S-8646. The APC was funded by MDPI.en_US
dc.identifier.doi10.3390/microorganisms12010150
dc.identifier.issn2076-2607
dc.identifier.issue1en_US
dc.identifier.pmid38257977en_US
dc.identifier.scopus2-s2.0-85183161755en_US
dc.identifier.scopusqualityQ2en_US
dc.identifier.urihttps://doi.org/10.3390/microorganisms12010150
dc.identifier.urihttps://hdl.handle.net/11411/8181
dc.identifier.volume12en_US
dc.identifier.wosWOS:001153064000001en_US
dc.identifier.wosqualityN/Aen_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherMdpien_US
dc.relation.ispartofMicroorganismsen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectPlasmid Dna Vaccinesen_US
dc.subjectPentose Phosphate Pathwayen_US
dc.subjectGlucose-Glycerol Co-Utilizationen_US
dc.subjectRecombinant Proteinen_US
dc.subjectMulticopy Plasmiden_US
dc.subjectMetabolic Burdenen_US
dc.subjectGrowth-Rateen_US
dc.subjectRecaen_US
dc.subjectFermentationen_US
dc.subjectStrainsen_US
dc.subjectVectoren_US
dc.subjectModelsen_US
dc.subjectAtpen_US
dc.titleIncreasing the Pentose Phosphate Pathway Flux to Improve Plasmid DNA Production in Engineered E. colien_US
dc.typeArticleen_US

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