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Implications for heterologous protein yield. Heat-shock and stringent responses have overlapping protease activity in Escherichia coli. Metabolic engineering for drug discovery and development. Metabolic engineering for the microbial production of 1,3-propanediol. Challenges in engineering microbes for biofuels production. These strategies should prove generalizeable to other metabolic pathways and programmable for fine-tuning pathway flux. One of the same scaffolds was used to triple the yield of glucaric acid, despite high titers (0.5 g/l) without the synthetic complex. The natural modularity of these domains enabled us to optimize the stoichiometry of three mevalonate biosynthetic enzymes recruited to a synthetic complex and thereby achieve 77-fold improvement in product titer with low enzyme expression and reduced metabolic load. Scaffolds bearing interaction domains from metazoan signaling proteins specifically accrue pathway enzymes tagged with their cognate peptide ligands. In an attempt to increase the effective concentration of each component of a pathway of interest, we built synthetic protein scaffolds that spatially recruit metabolic enzymes in a designable manner. Engineered metabolic pathways constructed from enzymes heterologous to the production host often suffer from flux imbalances, as they typically lack the regulatory mechanisms characteristic of natural metabolism.
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