Synthetic Transcription Factors: Detecting and Increasing the Production of Next Generation Bio-Based Chemicals and Fuels

Statement of Problem: In order to be able to develop in a robust manner, high-yield bioprocesses, especially for molecules that dont exist in nature, on a faster timescale and significant lower expenses then the current industry standard a multiplex, high-throughput screening platform for prospecting large pathway libraries is required. Technical Approach: Synthetic transcription factors, a technology developed at JBEI, has the potential to provide a general approach to this limitation and thereby enable the access of a wide variety of next generation biofuels. To demonstrate the general applicability of this technology to bio-based chemicals and fuels not found in nature and its integration into Arzedas core computational pathway and enzyme design technology stack, we will develop a high-throughput screen for levulinic acid, a promising novel building block for biorefineries, based on synthetic transcription factors. Phase I Work: We first establish the licensed technology from JBEI at Arzeda. We use this framework to construct synthetic transcription factors for levulinic acid. These synthetic transcription factors are then used to develop a multiplex, high-throughput pathway library platform that enables to rapid optimization of a high-yield levulinic acid producing bioprocess. Commercial Applications and Other Benefits: Levulinic acid is regarded to be one of the most attractive C5 building block as it can be converted to fuels, fuel additives, polymers and solvents. Arzedas high-yield fermentation route can produce levulinic acid at $0.75/lb and down to $0.5/lb, depending on the plant scale, enabling many of the above markets. On a societal level, this innovation will lead to a faster access to next generation biofuels and thereby help the nation reduce its dependence on foreign oil. Reports from the U.S. Office of Industrial Technologies estimate that levulinic acid at 200400M lb/yr scale could save 75.6 trillion Btu per year of energy by 2020. Finally, by providing a platform building block that can be produced economically with a fermentation process extremely similar to that of ethanol, the innovation could dramatically improve the economics of US-based biorefineries. Arzedas fermentation process to produce LA at low cost, high yield and large scale could ultimately help to protect US jobs, to decrease reliance on foreign oil and to secure the US lead in biotechnology Key Words: Synthetic biology, designer cell factory, next generation biofuel, levulinic acid, synthetic transcription factor, multiplex high-throughput screening of metabolic pathway libraries, computational pathway design, computational enzyme design Summary for Members of Congress: Currently, the development of high-yield bioprocesses for fuels is a time- and cost-intensive endeavor. Combining JBEIs and Arzedas technology we can enable the rapid optimization of economical, high-yield fermentation routes to next generation biofuels.