Light-based control of metabolic flux through assembly of synthetic organelles. Author Evan Zhao, Nathan Suek, Maxwell Wilson, Elliot Dine, Nicole Pannucci, Zemer Gitai, José Avalos, Jared Toettcher Publication Year 2019 Type Journal Article Abstract To maximize a desired product, metabolic engineers typically express enzymes to high, constant levels. Yet, permanent pathway activation can have undesirable consequences including competition with essential pathways and accumulation of toxic intermediates. Faced with similar challenges, natural metabolic systems compartmentalize enzymes into organelles or post-translationally induce activity under certain conditions. Here we report that optogenetic control can be used to extend compartmentalization and dynamic control to engineered metabolisms in yeast. We describe a suite of optogenetic tools to trigger assembly and disassembly of metabolically active enzyme clusters. Using the deoxyviolacein biosynthesis pathway as a model system, we find that light-switchable clustering can enhance product formation six-fold and product specificity 18-fold by decreasing the concentration of intermediate metabolites and reducing flux through competing pathways. Inducible compartmentalization of enzymes into synthetic organelles can thus be used to control engineered metabolic pathways, limit intermediates and favor the formation of desired products. Keywords Light, Saccharomyces cerevisiae, Optogenetics, Metabolic Networks and Pathways, Organelles, Metabolic Engineering, Synechocystis, Indoles, Synthetic Biology Journal Nat Chem Biol Volume 15 Issue 6 Pages 589-597 Date Published 2019 Jun ISSN Number 1552-4469 DOI 10.1038/s41589-019-0284-8 Alternate Journal Nat Chem Biol PMCID PMC6755918 PMID 31086330 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML