Media development and process parameter optimization using statistical experimental designs for the production of nonribosomal peptides in Escherichia coli |
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| Institution: | 1. Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, Germany;2. Faculty of Biology and Chemistry, Justus-Liebig University of Giessen, 35390 Giessen, Germany;3. Fraunhofer Institute for Molecular Biology and Applied Ecology, Project Group Bioresources, 35390 Giessen, Germany |
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| Abstract: | BackgroundNonribosomal peptide synthases (NRPS) can synthesize functionally diverse bioactive peptides by incorporating nonproteinogenic amino acids, offering a rich source of new drug leads. The bacterium Escherichia coli is a well-characterized production host and a promising candidate for the synthesis of nonribosomal peptides, but only limited bioprocess engineering has been reported for such molecules. We therefore developed a medium and optimized process parameters using the design of experiments (DoE) approach.ResultsWe found that glycerol is not suitable as a carbon source for rhabdopeptide production, at least for the NRPS used for this study. Alternative carbon sources from the tricarboxylic acid cycle achieved much higher yields. DoE was used to optimize the pH and temperature in a stirred-tank reactor, revealing that optimal growth and optimal production required substantially different conditions.ConclusionsWe developed a chemically defined adapted M9 medium matching the performance of complex medium (lysogeny broth) in terms of product concentration. The maximum yield in the reactor under optimized conditions was 126 mg L-1, representing a 31-fold increase compared to the first shaking-flask experiments with M9 medium and glycerol as the carbon source. Conditions that promoted cell growth tended to inhibit NRPS productivity. The challenge was therefore to find a compromise between these factors as the basis for further process development.How to cite: Oestreich AM, Suli LI, Gerlach D. et al. Media development and process parameter optimization using statistical experimental designs for the production of nonribosomal peptides in Escherichia coli. Electron J Biotechnol 2021;52. https://doi.org/10.1016/j.ejbt.2021.05.001 |
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| Keywords: | Alternative carbon source Bioprocess engineering Design of experiments Media development Microbial Biotechnology Nonribosomal peptide synthases pH optimization Rhabdopeptide Statistical experimental designs Temperature optimization |
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