Increasing the Pentose Phosphate Pathway Flux to Improve Plasmid DNA Production in Engineered E. coli
Küçük Resim Yok
Tarih
2024
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Mdpi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
The 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.
Açıklama
Anahtar Kelimeler
Plasmid Dna Vaccines, Pentose Phosphate Pathway, Glucose-Glycerol Co-Utilization, Recombinant Protein, Multicopy Plasmid, Metabolic Burden, Growth-Rate, Reca, Fermentation, Strains, Vector, Models, Atp
Kaynak
Microorganisms
WoS Q Değeri
N/A
Scopus Q Değeri
Q2
Cilt
12
Sayı
1
