STTR Phase I: Overcoming Metabolic Pathway Limitations through De Novo Pathway Design for Terpenoid Biosynthesis

这个小企业创新研究（STTR）第一阶段项目的目的是开发一种新的，高通量的萜类化合物前体途径，通过规避细菌甲基胆甾醇-磷酸（MEP）途径的限制，用于单萜类化合物的可再生生产。单萜类化合物是几种消费品的天然化学前体，许多是通过高污染的化学工艺生产的。在拟议的项目中，计划是通过设计从头代谢途径来回避一些MEP途径的限制。设计/预测的酶将被单独表征并组装成一个通路。此外，将使用多变量模块化代谢工程（MMME）方法来组装上游和下游途径，以优化代谢通量，从而在商业上可行的水平下过量生产。如果成功，该项目的更广泛影响/商业潜力将是开发一个利用可再生糖生产单萜类化合物的微生物生产平台，该平台将在美国保留和发展单萜类化合物衍生产品的可持续生产。通过这种策略，可以以比天然细菌MEP途径高得多的生产率制备萜类化合物。虽然目前的重点是10亿美元以上的单萜/衍生物市场，但这种方法将使美国所有萜类化合物的制造业受益，总共有50亿美元以上的市场。总的来说，该项目将为这些天然化学品提供一条新的可持续生产路线。

This Small Business Innovation Research (STTR) Phase I project aims to develop a novel, high flux terpenoid precursor pathway by circumventing limitations of the bacterial methyl erythritol-phosphate (MEP) pathway for the renewable production of monoterpenoids. Monoterpenoids are natural chemical precursors for several consumer products, and many are produced via highly polluting chemical processes. In the proposed project the plan is to sidestep some of the MEP pathway limitations by designing de novo metabolic pathways. The designed/predicted enzymes will be characterized individually and assembled into a pathway. Further, multivariate-modular metabolic engineering (MMME) approaches will be used to assemble the upstream and downstream pathways to optimize the metabolic flux for the overproduction at commercially viable levels. The broader impact/commercial potential of this project, if successful, will be to develop a microbial monoterpenoid production platform from renewable sugars that will retain and develop sustainable manufacturing of monoterpenoid-derived products in the US. By this strategy, terpenoids can be made at much higher productivities than the native bacterial MEP pathway. While the immediate focus is on the $1B+ monoterpene/derivative market, this approach will benefit US manufacturing of all terpenoids, in total a $5B+ market. Overall, this project will provide a new sustainable production route for these natural chemicals.