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

1403264/1403099 Wheeldon/Blenner该合作项目旨在开发一种先进的生物制造工艺，将废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、防腐涂料和香水的重要前体。目前，这些分子，长链二元酸，是从不可再生的石油原料中生产的，这些过程存在催化性能差和固有的安全问题。该项目致力于NSF促进国家健康、繁荣和福利的使命，通过创造一种内在安全的生化过程，该过程在低温和压力下运行，高效地将低成本原料转化为高价值产品，并创造新的可持续的生物制造技术。除了该项目的科学和工程目标外，教育推广活动还将把加利福尼亚州河滨县的研究生、本科生和社区大学生与南卡罗来纳州的学生联系起来。教育推广工作旨在增加STEM学生对先进生物和可持续制造研究的参与，从而满足南卡罗来纳州和加利福尼亚州河滨县对训练有素的STEM劳动力的迫切需求。这一过程利用某些酵母物种的天然能力，如解脂雅罗威酵母，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，有效地将自然产生的长链脂肪酸转化为二元酸。这些技术包括能够实现对基因表达的时间控制以及对生物合成途径活动的空间和时间控制的合成生物学工具。中心假设是，通过将氧化酶与脂质动员机制共同定位，可以提高氧化游离脂肪酸的生物合成途径的催化作用和产率。这一假说是基于实验和理论结果提出的，这些结果表明，通过酶共定位和细胞内途径定位，工程生物合成途径的产率提高。此外，对工程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途径的表达和途径通量的优化成为可能。开发的生化工艺专注于长链二元酸的生产，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其他生化过程，将脂肪转化为其他有用的化学物质，如高级生物燃料、短链二元酸和食品添加剂。该奖项由CBET分部生物技术、生化和生物质工程计划获得，由生物基础设施部生物研究仪器开发计划共同资助。

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.