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Metabolic engineering is a targeted and knowledge-based approach to improve production capabilities of microorganisms. It aims at increasing metabolic reaction rates towards product formation to obtain economic production processes. The fluxome, ie the computation of all metabolic reaction rates, is one cornerstone of metabolic engineering. For fluxomics, the study of intracellular reaction rates, several methods have been established. The most powerful one, 13C-metabolic flux analysis (13C-MFA), uses isotopically labeled substrates which are fed to cells. Emerging labeling patterns in the synthesized metabolites are measured by high-precision measurement devices like mass spectrometry. From the measured labeling pattern, the intracellular reaction rates can be estimated by mathematical modeling.

The present work faces 13C-MFA of complex systems. The non-model organism P. chrysogenum strain BCB1 is investigated in industrial environment with special focus on penicillin V production. The term “complex” is not only referring to the growth behavior of P. chrysogenum, but also includes side-product formation and compartmentalization of metabolism. This thesis aims at the transfer of 13C-MFA from a scientific application in a nearly ideal environment to an industrial standard. For this reason, the prerequisites needed and compromises made for 13C-MFA work-flow are thoroughly discussed and adaption of the industrial process is highlighted.