Synthetic biology for synthetic chemistry

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The progress in biotechnological disciplines such as metabolic engineering or synthetic biology increased the interest of chemical and pharmaceutical industries to implement microbial processes for chemical synthesis. However, most microorganisms, e. g., Escherichia coli or Saccharomyces cerevisiae, used in biotechnological applications are not evolved by nature for the production of industrially relevant compounds, which are often hydrophobic, non-charged, volatile, or toxic to the microbial organisms. Bioprocess design relies on an integrated approach addressing pathway, cellular, reaction, and process engineering to combine the results of natural evolution with the demands of industrial applicability. In this thesis, the microbial de novo production and selective oxyfunctionalization of the highly volatile isoprenoid limonene has been investigated as a model system featuring reactants with challenging physicochemical characteristics. Key constraints that limit limonene biosynthesis and its oxyfunctionalization in recombinant E. coli, related to genetics, physiology, and reaction engineering, were identified and relieved.