Metabolic analyses during fed-batch production of aromatic amino acids with Escherichia coli

Julia Tröndle, doctoral thesis Technical University of Munich, 2019

Parallelized short-term perturbation experiments with various substrates were employed to characterize the metabolism of recombinant E. coli from fed-batch processes for the production of aromatic amino acids by analyzing intracellular fluxes and metabolites. The expression of a transporter protein enabled also the conversion of a cell-internal substrate. Metabolic control analysis allowed the identification of limiting metabolic reactions during the production of L-tryptophan from glycerol.

Publications

  • Tröndle J, Schoppel K, Bleidt A, Trachtmann N, Sprenger GA, Weuster-Botz D (2020): Metabolic control analysis of Escherichia coli producing L-tryptophan from glycerol based on data from a parallelized short-term perturbation experiment. J Biotechnol 307: 15-28.
  • Tröndle J, Trachtmann N, Sprenger GA, Weuster-Botz D (2018): Production of L-tryptophan from glycerol using recombinant Escherichia coli. Biotechnol Bioeng 115: 2881–2892.
  • Tröndle J, Albermann C, Weiner M, Sprenger GA, Weuster-Botz D (2018): Phosphoenolpyruvate transporter enables targeted perturbation during metabolic analysis of L-phenylalanine production with Escherichia coli. Biotechnol J 13: 1700611.
  • Mairinger T, Tröndle J, Hanscho M, Hann S (2017): On-line clean-up and LC-MS analysis of primary metabolites in cell culture supernatants. Anal Methods 9: 5703-5710
  • Weiner M, Tröndle J, Albermann C, Sprenger GA, Weuster-Botz D (2017): Metabolic control analysis of L-phenylalanine production from glycerol with engineered E. coli using data from short-term steady-state perturbation experiments. Biochem Eng J 126: 86–100.
  • Weiner M, Tröndle J, Albermann C, Sprenger GA, Weuster-Botz D (2016): Perturbation experiments: Approaches for metabolic pathway analysis in bioreactors. Adv Biochem Eng Biotechnol 152: 91-136.
  • Weiner M, Tröndle J, Schmideder A, Binder K, Albermann C, Sprenger GA, Weuster-Botz D (2015): Parallelized small-scale production of uniformly 13C-labeled cell-extract for quantitative metabolome analysis, Anal Biochem 478: 134-140.
  • Albermann C, Weiner M, Tröndle J, Weuster-Botz D, Sprenger GA (2014): Utilization of organophosphate:phosphate antitransporter for isotope-labeling experiments in E. coli. FEMS Microbiol Lett 361: 52-61.
  • Weiner M, Tröndle J, Albermann C, Sprenger GA, Weuster-Botz D (2014): Carbon storage in recombinant Escherichia coli during growth on glycerol and lactic acid. Biotechnol Bioeng 111: 2508-2519.
  • Weiner M, Tröndle J, Albermann C, Sprenger GA, Weuster-Botz D (2014): Improvement of constraint-based flux estimation during L-phenylalanine production with Escherichia coli using targeted knock-out mutants. Biotechnol Bioeng 111: 1406-1416.