Utilisation of phosphorus nutrient content in industrial scale plasmid DNA production: A waste minimisation study

Human gene therapy is currently seeing an increase in the use of plasmid (pDNA)-based vectors as a preferred choice of vehicle for delivery of the therapeutic gene into the body. In this regard, the environmental impacts of the waste streams from the possible industrial scale manufacture of pDNA require more detailed assessment. In this study, an initial assessment was made of the nutrient phosphorus (P) inputs to four fermentation processes recommended for the industrial scale production of pDNA for the purposes of gene therapy. Phosphorus inputs to each of the four selected fermentation processes ranged from approximately 60 mg l^-1 up to 3000 mg l^-1 in the fresh media. However, the spent media waste from each of the processes exhibited only a minor reduction in the phosphorus concentrations indicating minimal uptake of P by the microorganisms. This unutilised excess level of phosphorus nutrient within the waste streams poses a strong potential for environmental impact. Waste minimisation studies were undertaken on one model fermentation process with the aim of reducing unnecessary phosphorus input. An optimised media containing a 98% reduction in added P to the media was developed. This phosphorus-minimised media had little quantitative effect upon cell biomass produced and no effect upon the quantity or quality of pDNA produced, relative to the control media. The reduction in P requirement results in an overall cost savings of 12% per fermentation batch, would simplify subsequent wastewater treatment and would contribute to slowing the depletion rate of a valuable, finite natural resource.