Enhanced production of plasmid DNA by engineered Escherichia coli strains
2012
Pablos, Tania E. | Soto, René | Mora, Eugenio Meza | Le Borgne, Sylvie | Ramírez, Octavio T. | Gosset, Guillermo | Lara, Alvaro R.
Escherichia coli strains VH33 (PTS⁻ GalP+ strain displaying a strongly reduced overflow metabolism) and VH34 (additionally lacking the pyruvate kinase A) were evaluated for the production of a plasmid DNA (pDNA) vaccine. The parent (W3110) and mutant strains were cultured using 10g of glucose/L. While the specific growth rates of the three strains were similar, they presented differences in the accumulation of acetate. W3110 accumulated up to 4g/L of acetate, VH33 produced 1.4g/L, and VH34 only 0.78g/L. VH33 and VH34 produced 76% and 300% more pDNA than W3110. Moreover, VH34 demanded 33% less oxygen than VH33 and W3110, which can be advantageous for large-scale applications.
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