In 2006, the FDA approved a plant cell culture process for production of the anticancer drug paclitaxel; however, suspension cultures have low product yields that are highly variable across cell lines and over time within a single cell line, leaving considerable room for process improvements. Superior paclitaxel-accumulating Taxus lines may be generated through stable over-expression of rate-influencing pathway genes or redirection of carbon flux away from competing metabolic pathways. We have established a reliable Agrobacterium-mediated stable transformation protocol for Taxus suspension cultures. The following parameters were found to be key for the consistent recovery of stable transformants: modification of antioxidants used during Agrobacterium infection, optimization of hygromycin concentrations in the selection medium and delayed transfer of infected cells to selection medium until there are signs of new callus growth. Utilizing this technology, we are implementing overexpression and silencing strategies to overcome bottlenecks to paclitaxel production by promoting carbon flux through the paclitaxel biosynthetic pathway.

Metabolic engineering of <i>Taxus</i> suspension cultures for enhanced production of paclitaxel

243rd National Meeting (2012)

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Metabolic engineering of Taxus suspension cultures for enhanced production of paclitaxel