Muconic Acid Production via Alternative Pathways and a Synthetic "metabolic Funnel"

Brian Thompson, Shawn Pugh, Michael Machas, David Nielsen

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Muconic acid is a promising platform biochemical and precursor to adipic acid, which can be used to synthesize various plastics and polymers. In this study, the systematic construction and comparative evaluation of a modular network of non-natural pathways for muconic acid biosynthesis was investigated in Escherichia coli, including via three distinct and novel pathways proceeding via phenol as a common intermediate. However, poor recombinant activity and high promiscuity of phenol hydroxylase ultimately limited "phenol-dependent" muconic acid production. A fourth pathway proceeding via p-hydroxybenzoate, protocatechuate, and catechol was accordingly developed, though with muconic acid titers by this route reaching just 819 mg/L, its performance lagged behind that of the established, "3-dehydroshikimiate-derived" route. Finally, these two most promising pathways were coexpressed in parallel to create a synthetic "metabolic funnel" that, by enabling maximal net precursor assimilation and flux while preserving native chorismate biosynthesis, nearly doubled muconic acid production to up to >3.1 g/L at a glucose yield of 158 mg/g while introducing only a single auxotrophy. This generalizable, "funneling" strategy is expected to have broad applications in metabolic engineering for further enhancing production of muconic acid, as well as other important bioproducts of interest.

Original languageEnglish (US)
Pages (from-to)565-575
Number of pages11
JournalACS Synthetic Biology
Volume7
Issue number2
DOIs
StatePublished - Feb 16 2018

Fingerprint

Acids
Phenols
Biosynthesis
phenol 2-monooxygenase
Phenol
Metabolic engineering
Metabolic Engineering
Escherichia coli
Plastics
muconic acid
Polymers
Glucose
Fluxes

Keywords

  • catechol
  • metabolic funnel
  • muconic acid
  • phenol

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Muconic Acid Production via Alternative Pathways and a Synthetic "metabolic Funnel". / Thompson, Brian; Pugh, Shawn; Machas, Michael; Nielsen, David.

In: ACS Synthetic Biology, Vol. 7, No. 2, 16.02.2018, p. 565-575.

Research output: Contribution to journalArticle

Thompson, Brian ; Pugh, Shawn ; Machas, Michael ; Nielsen, David. / Muconic Acid Production via Alternative Pathways and a Synthetic "metabolic Funnel". In: ACS Synthetic Biology. 2018 ; Vol. 7, No. 2. pp. 565-575.
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