Cell-Free Production of Terpenoid Chemicals and fules

A number of terpene-based chemicals could become renewable replacements for petroleum-based fuels. Yet so far, microbial conversion of biomass sugar to terpenes has not achieved the yields, titers and productivities needed to become competitive on price. Invizyne Technologies has developed cell-free systems that have the potential to achieve the necessary production parameters, but major advances will be required in nearly all aspects of cell-free processing to reach viable cost targets for low value chemicals like fuels. Techno-economic analysis identifies cofactors (NAD(P)+, NAD(P)H, ATP, Coenzyme A) as a major cost drivers for cell-free bioproduction. Indeed, the use of purified cofactors can contribute up to 85% of the total OPEX of a 5 MT/yr plant producing terpenoid biofuel from biomass derived sugars. In this Phase I application, we will develop a new process to isolate a cheaper crude cofactor mix from inexpensive waste yeast that can be employed for the low cost production of terpene-based chemicals. We expect to develop a process that can reduce cofactor costs by at least 10-fold, which in turn can decrease terpene production costs by greater than 75%. We will test a variety of lysis and cofactor isolation approaches on a small scale to identify the most effective methods that can be scaled safely and economically. We will also learn how to optimally incorporate the cofactor mixes into a cell free bioreactor system.Cell free bioprocessing has the potential to transform the production of renewable chemicals by lowering costs. The cell free approach could make rare, expensive chemicals more accessible for diverse applications. There could be great environmental benefits if cell free methods can make renewable high volume commodity chemicals and biofuels cost competitive with petroleum products.