This proposal describes the use of bacterial microcompartments (MCPs) to create a privileged pool of redox cofactors necessary for the high-yield production of biofuels and biochemicals from renewable carbon feedstocks. This unique approach addresses a key issue in biofuel production, namely, that biofuels are reduced molecules but intracellular redox pools are not favorable for reducing reactions, and necessary cellular processes compete for these limited pools. We propose to create custom MCPs with a private cofactor pool and recycling enzymes. This method permits segregation of redox cofactors from native cellular reactions, allowing for efficient reduction of encapsulated biofuels and biochemicals. Control over redox usage is a native feature of MCPs such as the 1,2-propanediol utilization and ethanolamine utilization MCPs, found in enteric Escherichia coli and Salmonella typhimurium strains. In these native systems, redox factor recycling is thought to be strictly coupled to the metabolic pathways that are also encapsulated. Our design will mimic this natural setup but with custom sets of enzymes that can be tailored for production of a variety of biofuels and biochemicals. An important element of our proposal is the creation of a kinetic metabolic model to explore the effect of compartmentalization on both the engineered pathway and native metabolism, which will be supported by metabolomics and transcriptomics data from our experimental work. The model will guide experimental design which in turn will guide the model, permitting us to study native microbial physiology for the organisms that employ these systems while simultaneously using the information to pioneer a new strategy for metabolic engineering and biofuel production. Our specific objectives are to: 1) Engineer and evaluate a 1,3-propanediol production MCP with private NADH pool 2) Engineer and evaluate C6 sugar catabolism MCP with private NADH pool 3) Establish kinetic metabolic models to explore link between compartmentalized reactions and cellular metabolism
|Effective start/end date||9/15/18 → 9/14/22|
- Department of Energy (DE-SC0019337 0002)
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