Motivation #1: Pathways removed from natural regulation when heterologously expressed

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Presentation transcript:

Problem We Address: Generalizeable Strategies for Improving Flux of Engineered Metabolic Pathways

Motivation #1: Pathways removed from natural regulation when heterologously expressed

Motivation #2: Differing levels of activity of pathway enzymes

Motivation #3: Complex Enzyme Structures Makes Direct Enzyme Optimization Difficult and Case-dependent

Our Strategy: Employ Natural Machinery for Targeted Pathway Regulation

Enzymes Have Different Levels of Activity (Turnover Fluxes)

Synthetic Scaffolds Can Co-localize Pathway Enzymes and Reduce Intermediate Runoff

Synthetic Scaffolds Can be Designed to Optimize Flux through the Pathway via Control of Relative Enzyme Ratios

Summary: Addition of Modularity To Nonmodular Metabolic Pathway Control over Pathway flux Through: 1. Corecruitment 2. Relative enzyme ratios

Synthetic Scaffolds Have Dramatic Effect On Model System (Mevalonate Pathway)

Modular Strategies are Scalable

Synopsis Modular parts (domain/ligands, enzymes) used to construct synthetic scaffolds are well-characterized. We are interested in how these parts can be used to build sophisticated machines and probing the engineering principles by which these machines can be programmed. Can predictive models be made based on simple equilibrium theory?