Auxin-mediated protein depletion for metabolic engineering in terpene-producing yeast

Nat Commun. 2021 Feb 16;12(1):1051. doi: 10.1038/s41467-021-21313-1.

Abstract

In metabolic engineering, loss-of-function experiments are used to understand and optimise metabolism. A conditional gene inactivation tool is required when gene deletion is lethal or detrimental to growth. Here, we exploit auxin-inducible protein degradation as a metabolic engineering approach in yeast. We demonstrate its effectiveness using terpenoid production. First, we target an essential prenyl-pyrophosphate metabolism protein, farnesyl pyrophosphate synthase (Erg20p). Degradation successfully redirects metabolic flux toward monoterpene (C10) production. Second, depleting hexokinase-2, a key protein in glucose signalling transduction, lifts glucose repression and boosts production of sesquiterpene (C15) nerolidol to 3.5 g L-1 in flask cultivation. Third, depleting acetyl-CoA carboxylase (Acc1p), another essential protein, delivers growth arrest without diminishing production capacity in nerolidol-producing yeast, providing a strategy to decouple growth and production. These studies demonstrate auxin-mediated protein degradation as an advanced tool for metabolic engineering. It also has potential for broader metabolic perturbation studies to better understand metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / metabolism
  • Cell Cycle Checkpoints / drug effects
  • Coenzyme A Ligases / metabolism
  • Glucose / metabolism
  • Hexokinase / metabolism
  • Indoleacetic Acids / pharmacology*
  • Limonene / metabolism
  • Metabolic Engineering*
  • Metabolic Flux Analysis
  • Polyisoprenyl Phosphates / metabolism
  • Proteolysis / drug effects
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Sesquiterpenes / metabolism
  • Terpenes / metabolism*

Substances

  • Bacterial Proteins
  • Indoleacetic Acids
  • Polyisoprenyl Phosphates
  • Saccharomyces cerevisiae Proteins
  • Sesquiterpenes
  • Terpenes
  • farnesyl pyrophosphate
  • Limonene
  • Hexokinase
  • Coenzyme A Ligases
  • malonyl-CoA synthetase
  • Glucose
  • nerolidol