(792b) Synthetic Co-Assembly of Metabolic Enzymes On Protein Scaffolds for Improved Flux

Enzyme complex formation is extremely common in natural metabolism.  In fact, recently been found that there are many more dynamic complexes dependent on environmental conditions than previously appreciated.  These enzyme-enzyme interactions can be important for allosteric regulation of catalytic activities, increased efficiency of intermediate hand-off, and subcellular localization.  Gaining an ability to engineer similar synthetic complexes in a programmable manner would provide several advantages to metabolic engineers for increasing pathway performance with lower enzyme expression levels, minimizing intermediate metabolite accumulation and resultant toxicity, and/or reducing undesired flux through side pathways.  We are taking a synthetic biology approach in co-localizing pathway enzymes into scaffolded complexes in an effort to probe the key design principles relating flux improvements achieved from formation of these complexes.  In particular, we are interested in testing our hypothesis that formation of multimeric complexes – an assembly of multiple scaffolded complexes – is critical to increased pathway performance.  I will also discuss some of the unexpected challenges we have encountered in building these scaffolded systems will be discussed.