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

该合作项目旨在开发一种先进的生物制造工艺，将低成本原料（如废甘油或生物质衍生糖）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，即长链二羧酸，是由不可再生的石油原料生产的，其工艺催化能力差，存在固有的安全问题。该项目通过创造一种内在安全的生物化学过程，在低温和低压下运行，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，为NSF推进国家健康、繁荣和福利的使命做出了贡献。除了这个项目的科学和工程目标之外，教育推广活动将把加州河滨县的研究生、本科生和社区学院的学生与南卡罗来纳州的学生联系起来。教育推广工作旨在提高STEM学生对先进生物和可持续制造研究的参与，从而解决南卡罗来纳州和加利福尼亚州里弗赛德县对训练有素的STEM劳动力的关键需求。这一过程利用了某些酵母菌的天然能力，如脂解耶氏酵母，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，有效地将自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，使基因表达的时间控制和生物合成途径活性的时空控制成为可能。核心假设是，通过将氧化酶与脂质动员机制共定位，可以提高游离脂肪酸氧化的生物合成途径的催化作用和产量。该假设是基于实验和理论结果制定的，这些结果表明通过酶共定位和细胞内途径定位可以提高工程生物合成途径的产量。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通路表达可调和通路通量优化成为可能。开发的生化工艺侧重于长链二羧酸的生产，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工艺，将脂质转化为其他有用的化学品，如先进的生物燃料、短链二羧酸和食品添加剂。该奖项由CBET部门的生物技术、生化和生物质工程项目颁发，由生物基础设施部门的生物研究仪器开发项目共同资助。

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.