Strain screening for isoprenoid production in bioreactors: balancing throughput and production scale constraints

To fulfill accelerating resource demands for a growing society, it will be necessary to achieve stable, environmentally sustainable and low-cost production of hydrocarbon-based fuels and chemicals. Amyris is accomplishing this by combining the power of synthetic biology with fermentation & downstream chemistry to engineer efficient microbial factories that manufacture isoprenoid products. Over 40,000 known compounds are derived from  the isoprenoid biosynthetic pathway, including molecules that are currently or might soon be used for biofuels, flavors, fragrances, steroids, sterols, emollients, coloring agents, pharmaceuticals, nutraceuticals, and monomers for polymer production. Using S. cerevisiae as a host, Amyris’ synthetic biology platform has accelerated creation and optimization of strains that produce high levels of isoprenoids. Automated strain engineering technology at Amyris allows us to create thousands of strains per week. These strains are screened in 96-well format via various high-throughput assays and the very best are promoted to be tested in 0.5L bioreactors. In the bioreactor, the production organisms are cultivated in a fed-batch mode and are for the first time subjected to the true production environment. The requirements of the screening process are twofold: it needs to be high throughput and also as representative of the 200,000L scale as is possible. This presentation will outline some major challenges at this step in the strain screening pipeline and how they were mitigated.