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

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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.