Engineering and application of a biosensor with focused ligand specificity
| Autor: | Della Corte, Dennis, van Beek, Hugo, Morris, Connor J., Grünberger, Alexander, Hofmann, Eckhard, Schröder, Gunnar F., Marienhagen, Jan, Syberg, Falk, Schallmey, Marcus, Tobola, Felix, Cormann, Kai, Schlicker, Christine, Baumann, Philipp Tobias, Krumbach, Karin, Sokolowsky, Sascha |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular Protein Conformation Science Biophysics Biosensing Techniques macromolecular substances Ligands Article Bacterial Proteins Protein Domains lcsh:Science Synthetic biology Crystallography Nanocrystallography Lysine technology industry and agriculture Microfluidic Analytical Techniques Flow Cytometry High-Throughput Screening Assays Corynebacterium glutamicum Metabolic Engineering Amino Acid Transport Systems Basic Thermodynamics lcsh:Q Molecular modelling ddc:500 |
| Zdroj: | Nature Communications, Vol 11, Iss 1, Pp 1-11 (2020) Nature Communications 11, 4851 (2020). doi:10.1038/s41467-020-18400-0 'Nature Communications ', vol: 11, pages: 4851-1-4851-11 (2020) Nature Communications Nature Communications 11(1), 4851 (2020). doi:10.1038/s41467-020-18400-0 |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/s41467-020-18400-0 |
| Popis: | Cell factories converting bio-based precursors to chemicals present an attractive avenue to a sustainable economy, yet screening of genetically diverse strain libraries to identify the best-performing whole-cell biocatalysts is a low-throughput endeavor. For this reason, transcriptional biosensors attract attention as they allow the screening of vast libraries when used in combination with fluorescence-activated cell sorting (FACS). However, broad ligand specificity of transcriptional regulators (TRs) often prohibits the development of such ultra-high-throughput screens. Here, we solve the structure of the TR LysG of Corynebacterium glutamicum, which detects all three basic amino acids. Based on this information, we follow a semi-rational engineering approach using a FACS-based screening/counterscreening strategy to generate an l-lysine insensitive LysG-based biosensor. This biosensor can be used to isolate l-histidine-producing strains by FACS, showing that TR engineering towards a more focused ligand spectrum can expand the scope of application of such metabolite sensors. Transcriptional biosensors represent powerful tools for the screening of vast strain libraries, but the broad ligand specificity of some transcriptional regulators (TRs) can prohibit such applications. Here authors present the engineering of a LysG-based biosensor with a focused ligand specificity to isolate L-histidine-producing strains. |
| Databáze: | OpenAIRE |
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