Rewriting yeast central carbon metabolism for industrial isoprenoid production

Nature. 2016 Sep 29;537(7622):694-697. doi: 10.1038/nature19769. Epub 2016 Sep 21.

Abstract

A bio-based economy has the potential to provide sustainable substitutes for petroleum-based products and new chemical building blocks for advanced materials. We previously engineered Saccharomyces cerevisiae for industrial production of the isoprenoid artemisinic acid for use in antimalarial treatments. Adapting these strains for biosynthesis of other isoprenoids such as β-farnesene (C15H24), a plant sesquiterpene with versatile industrial applications, is straightforward. However, S. cerevisiae uses a chemically inefficient pathway for isoprenoid biosynthesis, resulting in yield and productivity limitations incompatible with commodity-scale production. Here we use four non-native metabolic reactions to rewire central carbon metabolism in S. cerevisiae, enabling biosynthesis of cytosolic acetyl coenzyme A (acetyl-CoA, the two-carbon isoprenoid precursor) with a reduced ATP requirement, reduced loss of carbon to CO2-emitting reactions, and improved pathway redox balance. We show that strains with rewired central metabolism can devote an identical quantity of sugar to farnesene production as control strains, yet produce 25% more farnesene with that sugar while requiring 75% less oxygen. These changes lower feedstock costs and dramatically increase productivity in industrial fermentations which are by necessity oxygen-constrained. Despite altering key regulatory nodes, engineered strains grow robustly under taxing industrial conditions, maintaining stable yield for two weeks in broth that reaches >15% farnesene by volume. This illustrates that rewiring yeast central metabolism is a viable strategy for cost-effective, large-scale production of acetyl-CoA-derived molecules.

MeSH terms

  • Acetyl Coenzyme A / biosynthesis
  • Acetyl Coenzyme A / metabolism
  • Adenosine Triphosphate / metabolism
  • Bioreactors*
  • Biosynthetic Pathways
  • Carbohydrate Metabolism
  • Carbon / metabolism*
  • Carbon Dioxide / metabolism
  • Cytosol / metabolism
  • Fermentation
  • Metabolic Engineering*
  • Oxidation-Reduction
  • Oxygen / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / metabolism*
  • Sesquiterpenes / metabolism
  • Terpenes / metabolism*

Substances

  • Sesquiterpenes
  • Terpenes
  • Carbon Dioxide
  • beta-farnesene
  • Acetyl Coenzyme A
  • Carbon
  • Adenosine Triphosphate
  • Oxygen