Spatial control over enzyme organization presents a promising posttranslational strategy for improving metabolic flux. Directly tethering enzyme polypeptides has had inconsistent success. Use of a separate scaffold molecule, built from modular proteinprotein interaction domains, provides designable control over enzyme assembly parameters, including stoichiometry, as well as providing scalability for multiple enzymes. Thus, metabolic flux can be optimized by expression of these scaffolds in vivo. It is important to note that exploration of the use of synthetic scaffolds for improving metabolic flux is in its early stages. Accordingly, in this chapter, we describe efforts to date, hypotheses for scaffold function, and parameters to consider for application to new pathways. © 2011 Elsevier Inc. All rights reserved.
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