Incumbent routes to myriad high-value chemicals require energy-intensive manufacturing practices that are hazardous to human health and the environment, and non-renewable petrochemical feedstocks. Existing plant biomass conversion technologies in biofuel and bioproduct manufacturing are largely inefficient, preventing industrial scale-up of drop-in replacements and deployment for mass consumption. Lygos, Inc. proposes to develop a microbial catalyst that converts 100% renewable plant sugars into a ubiquitous, high-value bioproduct via a highly efficient biosynthetic pathway, thereby significantly increasing the landÂ’s effective product yield. (We cannot disclose the bioproduct due to the non- confidential nature of this summary.) Our proposed pathway further comprises CO2 sequestration, which increases theoretical yield of the bioproduct to >100%. We envision a bolt-on production facility co-located with cellulosic sugars and ethanol biofuel production. We anticipate upcycling waste CO2, thereby reducing industrial greenhouse gas emissions. Based on our techno-economic calculations, because the bio-product is more valuable than bio-ethanol, commercialization of the proposed technology in an integrated biorefinery can decrease biofuel production costs to < $3/GGE, thereby improving the economics of the overall facility. In this Phase I proposal, we plan to develop a prototype production strain capable of >5% bioproduct yield from cellulosic glucose and CO2. In summary, we plan to generate strain libraries comprising enzymes that constitute the pathway, and engineer strains towards improved pathway activity and yield. With in-house capability to construct 200 unique strains per month, we are able to carry out the work plan in the Phase I time period. By leveraging previous successes in bringing bio-malonic acid to market, we believe that a 5% yield provides sufficient demonstration of technical feasibility to de-risk the pathway enzymes. We anticipate Phase I results to enable a Phase II work plan that includes manufacturing at a 100-liter scale with established partners such as the DOE-funded Advanced Biofuels Process Development Unit. If the project is successful, we anticipate bringing to market a drop-in bio- product with a green, cost-advantaged process in a global market estimated to exceed $400MM.