Constructing recombinant Saccharomyces cerevisiae strains for malic-to-fumaric acid conversion

FEMS Microbiol Lett. 2023 Jan 17:370:fnad003. doi: 10.1093/femsle/fnad003.

Abstract

Saccharomyces cerevisiae with its robustness and good acid tolerance, is an attractive candidate for use in various industries, including waste-based biorefineries where a high-value organic acid is produced, such as fumaric acid could be beneficial. However, this yeast is not a natural producer of dicarboxylic acids, and genetic engineering of S. cerevisiae strains is required to achieve this outcome. Disruption of the natural FUM1 gene and the recombinant expression of fumarase and malate transporter genes improved the malic acid-to-fumaric acid conversion by engineered S. cerevisiae strains. The efficacy of the strains was significantly influenced by the source of the fumarase gene (yeast versus bacterial), the presence of the XYNSEC signal secretion signal and the available oxygen in synthetic media cultivations. The ΔFUM1Ckr_fum + mae1 and ΔFUM1(ss)Ckr_fum + mae1 strains converted extracellular malic acid into 0.98 and 1.11 g/L fumaric acid under aerobic conditions.

Keywords: Saccharomyces cerevisiae; fumarase; fumaric acid; malate transporter; malic acid; organic acids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fumarate Hydratase* / genetics
  • Fumarate Hydratase* / metabolism
  • Malates / metabolism
  • Saccharomyces cerevisiae* / genetics
  • Saccharomyces cerevisiae* / metabolism

Substances

  • malic acid
  • fumaric acid
  • Fumarate Hydratase
  • Malates