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

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Problem We Address Generalizeable Strategies
for Improving Flux of Engineered Metabolic
Pathways
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Motivation 1Pathways removed from natural
regulation when heterologously expressed
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Motivation 2Differing levels of activity of
pathway enzymes
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Motivation 3Complex Enzyme Structures Makes
Direct Enzyme Optimization Difficult and
Case-dependent
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Our Strategy Employ Natural Machinery for
Targeted Pathway Regulation
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Enzymes Have Different Levels of Activity
(Turnover Fluxes)
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Synthetic Scaffolds Can Co-localize Pathway
Enzymes and Reduce Intermediate Runoff
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Synthetic Scaffolds Can be Designed to Optimize
Flux through the Pathway via Control of Relative
Enzyme Ratios
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Summary Addition of ModularityTo Nonmodular
Metabolic Pathway
Control over Pathway flux Through 1.
Corecruitment 2. Relative enzyme ratios
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Synthetic Scaffolds Have Dramatic Effect On Model
System (Mevalonate Pathway)
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Modular Strategies are Scalable
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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?