title: Robust identification of metabolic control for microbial l-methionine production following an easy-to-use puristic approach.
abstract: The identification of promising metabolic engineering targets is a key issue in metabolic control analysis (MCA). Conventional approaches make intensive use of model-based studies, such as exploiting post-pulse metabolic dynamics after proper perturbation of the microbial system. Here, we present an easy-to-use, purely data-driven approach, defining pool efflux capacities (PEC) for identifying reactions that exert the highest flux control in linear pathways. Comparisons with linlog-based MCA and data-driven substrate elasticities (DDSE) showed that similar key control steps were identified using PEC. Using the example of l-methionine production with recombinant Escherichia coli, PEC consistently and robustly identified main flux controls using perturbation data after a non-labeled12C-l-serine stimulus. Furthermore, the application of full-labeled13C-l-serine stimuli yielded additional insights into stimulus propagation to l-methionine. PEC analysis performed on the13C data set revealed the same targets as the12C data set. Notably, the typical drawback of metabolome analysis, namely, the omnipresent leakage of metabolites, was excluded using the13C PEC approach.
authors.affiliation: Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Teleki
authors.affiliation: Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Rahnert
authors.affiliation: Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
authors.forename: O
authors.initials: O
authors.lastname: Bungart
authors.affiliation: Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Gann
authors.affiliation: Evonik Nutrition & Care GmbH, Kantstr. 2, 33790 Halle, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Ochrombel
authors.affiliation: Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. Electronic address: ralf.takors@ibvt.uni-stuttgart.de.
authors.forename: R
authors.initials: R
authors.lastname: Takors
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D055442:Metabolome / Q000235:genetics*; D008715:Methionine / Q000096:biosynthesis / Q000235:genetics; D008954:Models, Biological
chemical_list: D008715:Methionine
keywords: Metabolic control analysis; Quantitative metabolomics; Stimulus response experiment