Enhanced production of plasmid DNA by engineered Escherichia coli strains |
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Authors: | Pablos Tania E Soto René Mora Eugenio Meza Le Borgne Sylvie Ramírez Octavio T Gosset Guillermo Lara Alvaro R |
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Affiliation: | Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Artificios No. 40, Col. Miguel Hidalgo, Del. álvaro Obregón, México DF, CP 01120, Mexico. |
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Abstract: | 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 10 g 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 4 g/L of acetate, VH33 produced 1.4 g/L, and VH34 only 0.78 g/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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