Elsevier

Metabolic Engineering

Volume 31, September 2015, Pages 181-188
Metabolic Engineering

Establishment of a yeast platform strain for production of p-coumaric acid through metabolic engineering of aromatic amino acid biosynthesis

https://doi.org/10.1016/j.ymben.2015.08.003Get rights and content
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Highlights

  • p-Coumaric acid (CA) is a precursor for biosynthesis of flavonoids, polyketides, etc.

  • We engineered a CA-overproducing Saccharomyces cerevisiae platform strain.

  • Nearly 2 g L−1 CA were produced from glucose in fed-batch fermentation.

  • Shikimate kinase is one of the flux-controlling steps in aromatic amino acid biosynthesis.

Abstract

Aromatic amino acids are precursors of numerous plant secondary metabolites with diverse biological functions. Many of these secondary metabolites are already being used as active pharmaceutical or nutraceutical ingredients, and there are numerous exploratory studies of other compounds with promising applications. p-Coumaric acid is derived from aromatic amino acids and, besides being a valuable chemical building block, it serves as precursor for biosynthesis of many secondary metabolites, such as polyphenols, flavonoids, and some polyketides.

Here we developed a p-coumaric acid-overproducing Saccharomyces cerevisiae platform strain. First, we reduced by-product formation by knocking out phenylpyruvate decarboxylase ARO10 and pyruvate decarboxylase PDC5. Second, different versions of feedback-resistant DAHP synthase and chorismate mutase were overexpressed. Finally, we identified shikimate kinase as another important flux-controlling step in the aromatic amino acid pathway by overexpressing enzymes from Escherichia coli, homologous to the pentafunctional enzyme Aro1p and to the bifunctional chorismate synthase-flavin reductase Aro2p. The highest titer of p-coumaric acid of 1.93±0.26 g L−1 was obtained, when overexpressing tyrosine ammonia-lyase TAL from Flavobacterium johnsoniaeu, DAHP synthase ARO4K229L, chorismate mutase ARO7G141S and E. coli shikimate kinase II (aroL) in Δpdc5Δaro10 strain background. To our knowledge this is the highest reported titer of an aromatic compound produced by yeast.

The developed S. cerevisiae strain represents an attractive platform host for production of p-coumaric-acid derived secondary metabolites, such as flavonoids, polyphenols, and polyketides.

Keywords

p-Coumaric acid
Saccharomyces cerevisiae
Shikimate kinase

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