Combining metabolic engineering and synthetic biology to engineer natural product synthesis in oleaginous yeast

植物产生具有药用特性的化学物质。由于浓度低，从天然来源中分离植物天然产物（PNP）是困难的。由于PNP的结构复杂性，化学合成通常不经济。该项目将修改酵母生产具有抗癌特性的PNP。该分子代表了一类具有治疗价值的PNP。如果酵母可以产生高浓度的PNP，这些药物可能会变得更便宜。还计划开展外联活动。研究机会将提供给少数民族本科生。教学示范和科学博览会介绍将在巴尔的摩和华盛顿地区。这些努力的目标是招募女高中生和代表性不足的少数民族从事STEM职业。为了提高PNP生产的成本竞争力，该项目将解决辅因子平衡（ATP，NADPH，CoASH等），并最大限度地提高碳流和电子还原效率。这种跨学科的方法结合了基因工程，基因调控，适应性实验室进化，耐受性表型，代谢谱和细胞区室化工程。生物化学和遗传策略将消除HMG-CoA途径负反馈控制，改善代谢物运输，并过表达植物P450酶。 这些努力应提高途径效率，并使PNPs的化学结构多样化。我们的战略将针对途径产量和总制造成本。我们的目标是建立一个具有成本竞争力的环状三萜类生物制造平台，大大降低制造成本。该项目将为工程Y.该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Plants produce chemicals with medicinal properties. Isolating plant natural products (PNPs) from native sources is difficult due to low concentrations. Chemical synthesis is often not economical because of the structural complexity of PNPs. This project will modify yeast to produce a PNP with anti-cancer properties. This molecule represents a class of PNPs with therapeutic value. If yeast can produce high concentrations of PNPs, these drugs could become more affordable. Outreach activities are also planned. Research opportunities will be offered to minority undergraduates. Teaching demonstrations and science fair presentations will be made in Baltimore and the DC area. The goal of these efforts will be to recruit female high school students and underrepresented minorities to pursue STEM careers. To improve the cost-competitiveness of PNP production, the project will address cofactor balance (ATP, NADPH, CoASH et al) and maximize the carbon flow and electron reduction efficiency towards the target pathway. This interdisciplinary approach combines genetic engineering, gene regulation, adaptive lab evolution, tolerance phenotyping, metabolic profiling, and cellular compartmentalization engineering. Biochemical and genetic strategies will abolish HMG-CoA pathway negative feedback control, improve metabolite trafficking, and overexpress plant P450 enzymes. These efforts should improve pathway efficiency and diversify the chemical structure of PNPs. Our strategy will target pathway yield and the total cost of manufacturing. The goal is to build a cost-competitive cyclic triterpenoids biomanufacturing platform with significantly reduced manufacturing cost. This project will provide a testbed for engineering Y. lipolytica and expanding nature's capacity to produce complex PNPs with pharmaceutical values for clinical trials and treating emerging diseases.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.