(176v) Resource Mapping and Model-Informed Strain Design to Enhance pDNA Productivity in Bacterial Systems

The rise in demand for plasmid DNA (pDNA) and anticipated increases in market size prompt examination of how this material, important to both fundamental research and therapeutic applications is manufactured. Traditionally, common Escherichia coli strains have served in this capacity with media optimization key to most optimizations. Spaces that may be explored remain and include strategic alterations to the host cell that increase productivity but not at the cost of fidelity, and choosing a strain other than E. coli to complement medium design. This poster will describe our first foray into the field of pDNA manufacture, a combination of mathematical and experimental characterization of E. coli, Vibrio natriegens, and several lactic acid bacteria. Constraint-based flux analysis incorporating pDNA composition, size, and central metabolism was used to characterize growth and plasmid yield, and with this information in hand changes to strain(s) have been examined. Results indicate that order of magnitude changes in yield are feasible and with such an increase may compete favorably against traditional strains like E. coli DH5 alpha.