Synthetic protein scaffolds provide modular control over metabolic flux Export

@article{citeulike:5389413, abstract = {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. 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. Scaffolds bearing interaction domains from metazoan signaling proteins specifically accrue pathway enzymes tagged with their cognate peptide ligands. 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. 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. These strategies should prove generalizeable to other metabolic pathways and programmable for fine-tuning pathway flux.}, author = {Dueber, John E. and Wu, Gabriel C. and Malmirchegini, G. Reza and Moon, Tae S. and Petzold, Christopher J. and Ullal, Adeeti V. and Prather, Kristala L. J. and Keasling, Jay D.}, citeulike-article-id = {5389413}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nbt.1557}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nbt.1557}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/19648908}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=19648908}, day = {02}, doi = {10.1038/nbt.1557}, issn = {1087-0156}, journal = {Nature Biotechnology}, keywords = {igem, metabolic\_channeling, protein\_scaffolds}, month = {August}, number = {8}, pages = {753--759}, posted-at = {2009-08-16 03:39:37}, priority = {2}, publisher = {Nature Publishing Group}, title = {Synthetic protein scaffolds provide modular control over metabolic flux}, url = {http://dx.doi.org/10.1038/nbt.1557}, volume = {27}, year = {2009} }

TY - JOUR ID - citeulike:5389413 L3 - citeulike-article-id:5389413 N2 - 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. 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. Scaffolds bearing interaction domains from metazoan signaling proteins specifically accrue pathway enzymes tagged with their cognate peptide ligands. 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. 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. These strategies should prove generalizeable to other metabolic pathways and programmable for fine-tuning pathway flux. IS - 8 JF - Nature Biotechnology SN - 1087-0156 EP - 759 TI - Synthetic protein scaffolds provide modular control over metabolic flux PB - Nature Publishing Group VL - 27 SP - 753 KW - igem KW - metabolic_channeling KW - protein_scaffolds AU - Dueber, John AU - Wu, Gabriel AU - Malmirchegini, Reza AU - Moon, Tae AU - Petzold, Christopher AU - Ullal, Adeeti AU - Prather, Kristala AU - Keasling, Jay PY - 2009/08/02/ UR - http://dx.doi.org/10.1038/nbt.1557 ER -