Rapid Media Transition for Metabolic Control Analysis of Fed-Batch Fermentation Processes

Hannes Link, doctoral thesis Technische Universität München, 2009

Identification of limiting capacities in metabolism of a producer strain is currently not possible in biotechnological production processes. In this work, a new experimental approach was developed to separate micro-organisms during a bioprocess and analyse them in a bioreactor operated in parallel. In short-term scale experiments of several minutes metabolic disturbances are induced in the perturbation reactor and the biological response is captured quantitatively by metabolic flux and metablome analyses. Using kinetic modeling of intracellular metabolism and sensitivity analysis, the approach revealed for the first time the control structure of central metabolism of Escherichia coli under conditions of a fed-batch process.


  • Link H, Anselment B, Weuster-Botz D (2010): Rapid media transition: An experimental approach for steady state analysis of metabolic pathways. Biotechnol Prog 26: 1-10.
  • Link H, Anselment B, Weuster-Botz D (2008): Leakage of adenylates during cold methanol/glycerol quenching of Escherichia coli. Metabolomics 4: 240-247.
  • Link H, Vera J, Weuster-Botz D, Torres Darias N, Franco-Lara E (2008): Multi-objective steady state optimization of biochemical reaction networks using a constrained genetic algorithm. Comp Chem Eng 32: 1707-1713.
  • Link H, Weuster-Botz D (2007): Steady-state analysis of metabolic pathways: Comparing the double modulation method and the lin-log approach. Metab Eng 9: 433-441.

Further Publications

  • Link H, Weuster-Botz D (2011): Upstream Processing: Medium Formulation and Development. In Murray Moo-Young (ed.): Comprehensive Biotechnology, Second Edition, Vol. 2. Elsevier: 119-134.
  • Braun M, Link H, Liu L, Schmid RD, Weuster-Botz D (2011): Biocatalytic process optimization based on mechanistic modeling of cholic acid oxidation with cofactor regeneration. Biotechnol Bioeng 108: 1307-1317.
  • Tiemeyer A, Link H, Weuster-Botz D (2007): Kinetic studies on autohydrogenotrophic growth of Ralstonia eutropha with nitrate as terminal electron acceptor. Appl Microbiol Biotechnol 76: 75-81.
  • Link H, Weuster-Botz D (2006): Genetic algorithm for multi-objective experimental optimization. Bioprocess Biosyst Eng 29: 385-390.
  • Franco-Lara E, Link H, Weuster-Botz D (2006): Evaluation of artificial neural networks for modelling and optimization of medium composition with a genetic algorithm. Proc Biochem 41: 2200-2206.
  • Havel J, Link H, Hofinger M, Franco-Lara E, Weuster-Botz D (2006): Comparison of genetic algorithms for experimental multi-objective optimization on the example of medium design for cyanobacteria. Biotechnol J 1: 549-555.
  • Amidjojo A, Franco-Lara E, Nowak A, Link H, Weuster-Botz D (2005): Asymmetric synthesis of tert-butyl (3R, 5S) 6-chlorodihydroxyhexanoate with Lactobacillus kefir. Appl Microbiol Biotechnol 69: 9-15.