High Kanamycin Concentration as Another Stress Factor Additional to Temperature to Increase pDNA Production in E. coli DH5α Batch and Fed-Batch Cultures.

摘要: Plasmid DNA (pDNA) vaccines require high supercoiled-pDNA doses (milligrams) to achieve an adequate immune response. Therefore, processes development obtain pDNA yields and productivity is crucial. production affected by several factors including culture type, medium composition, growth conditions. We evaluated the effect of kanamycin concentration temperature on production, overflow metabolism (organic acids) metabolic burden (neomycin phosphotransferase II) in batch fed-batch cultures Escherichia coli DH5α-pVAX1-NH36. Results indicated that increases volumetric productivity, specific when were carried out at 42 °C, reduced but increased. Micrographs taken with a scanning electron microscope (SEM) analyzed, showing important morphological changes. The (300 mg/L) was cell density (50 gDCW/L), which reached using increase controlling heating rates. yield 759 mg/L 31.19 mg/L/h, respectively, two-fold greater than control 50 mg/L. A stress-based process simultaneously caused can be successfully applied production.