Dynamic Metabolic Control for the Production of Limonene

Biorefining is the sustainable processing of biomass into a spectrum of marketable products such as fuels, power, heat, food, feed and value-added chemicals. By producing multiple products, a bio-refinery takes advantage of the various components in biomass and their intermediates therefore maximizing the value derived from the biomass feedstock. A bio-refinery could, for example, produce one or several low-volume, but high-value, chemical ornutraceuticalproducts and a low-value, but high-volume liquid transportation fuel such asbiodieselor bioethanol. At the same time generating electricity and process heat, throughcombined heat and power(CHP) technology, for its own use and perhaps enough for sale of electricity to the local utility. The high-value products increase profitability, the high-volume fuel helps meet energy needs, and the power production helps to lower energy costs and reduce greenhouse gasemissions from traditional power plant facilities. Although some facilities exist that can be called bio-refineries, the complete bio-refinery concept has yet to be fully realized.There are a number of significant barriers to the realization of self-sufficient bio-refineries. In addition to the technological challenges, factors such as political and market resistance, lack of supporting infrastructure (including feedstock logistics), and market development all must be addressed. One of the most challenging technological barriers to date for biochemical conversion technologies (such as those that use engineered microbes and fermentation) is that each bio-process that has been developed is unique and requires a customized process and related process equipment. In addition, due to the complex responses of microbial systems to environmental variables, routine predictable scale-up from small-scale screening data has remained elusive to the field. As result, the Federal Government's initiatives to develop a multi-product bio-refinery utilizing agricultural carbohydrate feedstock have yet to be realized.Our Dynamic Metabolic Control platform provides an innovative and disruptiveapproach to rapidly commercialize bio-based products. This technology provides for the generation of robust microbial hosts with unparalleled reproducibility across scale, from micro-well to commercial production. Importantly, we use a single, standardized 2-stage bioprocess for the production of broad diversity of products. These features directly contribute to overcoming one of the key technology barriers in the realization of the bio-refinery concept.The proposed program directly supports the USDA strategic goal of promoting agriculturally-related manufacturing technology in the area of alternative and renewable energy. Support of this proposed program by USDA, along with success in related in-progress programs would significantly advance progress toward realization of the first integrated bio-refinery.