Production of itaconic acid from alkali pretreated lignin by dynamic two stage bioconversion

Nat Commun. 2021 Apr 15;12(1):2261. doi: 10.1038/s41467-021-22556-8.

Abstract

Expanding the portfolio of products that can be made from lignin will be critical to enabling a viable bio-based economy. Here, we engineer Pseudomonas putida for high-yield production of the tricarboxylic acid cycle-derived building block chemical, itaconic acid, from model aromatic compounds and aromatics derived from lignin. We develop a nitrogen starvation-detecting biosensor for dynamic two-stage bioproduction in which itaconic acid is produced during a non-growth associated production phase. Through the use of two distinct itaconic acid production pathways, the tuning of TCA cycle gene expression, deletion of competing pathways, and dynamic regulation, we achieve an overall maximum yield of 56% (mol/mol) and titer of 1.3 g/L from p-coumarate, and 1.4 g/L titer from monomeric aromatic compounds produced from alkali-treated lignin. This work illustrates a proof-of-principle that using dynamic metabolic control to reroute carbon after it enters central metabolism enables production of valuable chemicals from lignin at high yields by relieving the burden of constitutively expressing toxic heterologous pathways.

Publication types

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

MeSH terms

  • Alkalies / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Basidiomycota / enzymology
  • Basidiomycota / genetics
  • Biosensing Techniques
  • Burkholderia / enzymology
  • Burkholderia / genetics
  • Carbon / metabolism
  • Citric Acid Cycle / genetics
  • Coumaric Acids / metabolism
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Industrial Microbiology / methods
  • Lignin / chemistry
  • Lignin / metabolism*
  • Metabolic Engineering / methods*
  • Proof of Concept Study
  • Pseudomonas putida / genetics
  • Pseudomonas putida / metabolism*
  • Succinates / metabolism*

Substances

  • Alkalies
  • Bacterial Proteins
  • Coumaric Acids
  • Fungal Proteins
  • Succinates
  • Carbon
  • Lignin
  • p-coumaric acid
  • itaconic acid

Supplementary concepts

  • Ustilago maydis