Collaborative Research: Intracellular localization of biosynthetic pathways for conversion of lipids to dicarboxylic acids in oleaginous yeast

1403264/1403099Wheeldon/Blenner 该合作项目旨在开发一种先进的生物制造工艺，将废弃甘油或生物质衍生糖等低成本原料转化为特种聚合物、粘合剂、防腐涂料和香料的重要前体。目前，这些分子，长链二羧酸，是由不可再生的石油原料在遭受不良催化和固有安全问题的过程中生产的。该项目通过创建一种在低温和低压下运行的内在安全的生化过程，有助于实现NSF推进国家健康，繁荣和福利的使命，有效地将低成本原料转化为高价值产品，并创造新的可持续生物制造技术。除了这个项目的科学和工程目标，教育推广活动将连接研究生，本科生和社区学院的学生在滨江县，加利福尼亚州与学生在南卡罗来纳州。教育推广工作旨在增加STEM学生参与先进的生物和可持续制造研究，从而满足南卡罗来纳州和滨江县对训练有素的STEM劳动力的迫切需求。该过程利用某些酵母物种的天然能力，如酵母解脂耶氏酵母，代谢甘油和糖，并产生高产量的长链脂肪酸。该项目开发的创新技术将允许在活性酵母细胞内发生协调反应，以有效地将天然产生的长链脂肪酸转化为二羧酸。这些技术包括合成生物学工具，能够对基因表达进行时间控制，并对生物合成途径活性进行空间和时间控制。中心假设是，用于游离脂肪酸氧化的生物合成途径的催化和产率可以通过将氧化酶与脂质动员机制共定位来增强。该假设是根据实验和理论结果制定的，这些结果表明，通过酶共定位和通过细胞内途径定位，工程生物合成途径的产量增加。此外，对工程化途径的脂肪酸底物敏感的基因调控元件的开发和实施将使得能够实现可调节的途径表达和途径通量的优化。已开发的生物化学工艺专注于生产长链二羧酸，但预计成功创建该工艺所需的创新合成生物学工具将广泛适用于将脂质转化为其他有用化学品（如先进生物燃料、短链二羧酸和食品添加剂）的其他生物化学工艺。CBET部门的生物质工程项目由生物基础设施部门的生物研究项目仪器开发共同资助。

1403264/1403099Wheeldon/Blenner This collaborative project aims to develop an advanced bio-manufacturing process to convert low cost feedstocks such as waste glycerol or biomass-derived sugars into important precursors for specialty polymers, adhesives, anti-corrosive coatings, and fragrances. Currently these molecules, long chain dicarboxylic acids, are produced from non-renewable, petroleum feedstocks in processes that suffer from poor catalysis and inherent safety concerns. This project contributes towards NSF's mission of advancing national heath, prosperity, and welfare by creating an inherently safe biochemical process that operates at low temperature and pressures, is efficient in the conversion of low cost feedstocks into high value products, and creates new sustainable bio-manufacturing technologies. In addition to the scientific and engineering goals of this project, educational outreach activities will connect graduate, undergraduate, and community college students in Riverside County, CA with students in South Carolina. The educational outreach efforts aim to increase participation of STEM students in advanced bio- and sustainable-manufacturing research, thus addressing critical needs in both South Carolina and Riverside County, CA for well-trained STEM workforces.The process exploits the natural abilities of certain yeast species, such as the yeast Yarrowia lipolytica, to metabolize glycerol and sugars, and produce high yields of long chain fatty acids. The innovative technologies developed in this project will allow for coordinated reactions to occur inside active yeast cells to efficiently convert the naturally produced long chain fatty acids into dicarboxylic acids. These technologies include synthetic biology tools that enable the temporal control of gene expression and the spatial and temporal control of biosynthetic pathway activity. The central hypothesis is that the catalysis and yield of biosynthetic pathways for the oxidation of free fatty acids can be enhanced by co-localizing oxidative enzymes with the lipid mobilization machinery. The hypothesis was formulated based on experimental and theoretical results that demonstrate enhanced yields of engineered biosynthetic pathways via enzyme co-localization and via intracellular pathway localization. Moreover, the development and implementation of gene regulatory elements sensitive to the fatty acid substrates of the engineered pathway will enable tunable pathway expression and optimization of pathway flux. The developed biochemical process focuses on the production of long chain dicarboxylic acids, but it is anticipated that the innovative synthetic biology tools needed to successfully create the process will be broadly applicable to other biochemical processes for the conversion of lipids into other useful chemicals such as advanced biofuels, short chain dicarboxlyic acids, and food additives.This award by the Biotechnology, Biochemical, and Biomass Engineering Program of the CBET Division is co-funded by the Instrument Development for Biological Research Program of the Division of Biological Infrastructure.