Light-based control of metabolic flux through assembly of synthetic organelles

Nat Chem Biol. 2019 Jun;15(6):589-597. doi: 10.1038/s41589-019-0284-8. Epub 2019 May 13.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Indoles / metabolism
  • Light*
  • Metabolic Engineering*
  • Metabolic Networks and Pathways / radiation effects*
  • Optogenetics / methods*
  • Organelles / chemistry
  • Organelles / metabolism*
  • Organelles / radiation effects*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / radiation effects
  • Synechocystis / radiation effects
  • Synthetic Biology*

Substances

  • Indoles
  • violacein