Intracellular Flux Prediction of Recombinant Escherichia coli Producing Gamma-Aminobutyric Acid

J Microbiol Biotechnol. 2024 Apr 28;34(4):978-984. doi: 10.4014/jmb.2312.12022. Epub 2024 Jan 30.

Abstract

Genome-scale metabolic model (GEM) can be used to simulate cellular metabolic phenotypes under various environmental or genetic conditions. This study utilized the GEM to observe the internal metabolic fluxes of recombinant Escherichia coli producing gamma-aminobutyric acid (GABA). Recombinant E. coli was cultivated in a fermenter under three conditions: pH 7, pH 5, and additional succinic acids. External fluxes were calculated from cultivation results, and internal fluxes were calculated through flux optimization. Based on the internal flux analysis, glycolysis and pentose phosphate pathways were repressed under cultivation at pH 5, even though glutamate dehydrogenase increased GABA production. Notably, this repression was halted by adding succinic acid. Furthermore, proper sucA repression is a promising target for developing strains more capable of producing GABA.

Keywords: Escherichia coli; Genome-scale metabolic model; fermentation; gamma-aminobutyric acid.

MeSH terms

  • Bioreactors / microbiology
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Fermentation
  • Glutamate Dehydrogenase / genetics
  • Glutamate Dehydrogenase / metabolism
  • Glycolysis
  • Hydrogen-Ion Concentration
  • Metabolic Engineering / methods
  • Metabolic Flux Analysis
  • Models, Biological
  • Pentose Phosphate Pathway
  • Succinic Acid / metabolism
  • gamma-Aminobutyric Acid* / biosynthesis
  • gamma-Aminobutyric Acid* / metabolism

Substances

  • gamma-Aminobutyric Acid
  • Succinic Acid
  • Glutamate Dehydrogenase