Metabolic Control Analysis of microbial fed-batch production of L-phenylalanine

Michael Weiner, doctoral thesis Technische Universität München, 2015

Metabolic disturbances were induced in recombinant E. coli cells producing L-phenylalanine by using different substrates in parallelized stationary short-term (20 minutes) perturbation experiments in stirred-tank bioreactors. Cellular response and deflection of metabolism were monitored by analyses of fluxome and metabolome. Identification of metabolic control of L-phenylalanine production with glycerol was enabled at several points of the production process by applying kinetic network models.

Publications

• 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, DOI: 10.1002/biot.201700611.
• 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.
• Weiner M, Albermann C, Gottlieb K, Sprenger GA, Weuster-Botz D (2014): Fed-batch production of L-phenylalanine from glycerol and ammonia with recombinant Escherichia coli. Biochem Eng J 83: 62-69.