> Details

Della Corte, Dennis; van Beek, Hugo L.; Syberg, Falk; Schallmey, Marcus; Tobola, Felix; Cormann, Kai U.; Schlicker, Christine; Baumann, Philipp T.; Krumbach, Karin; Sokolowsky, Sascha; Morris, Connor J.; Grünberger, Alexander; Hofmann, Eckhard; Schröder, Gunnar F.; Marienhagen, Jan

Engineering and application of a biosensor with focused ligand specificity

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  • Media type: E-Article
  • Title: Engineering and application of a biosensor with focused ligand specificity
  • Contributor: Della Corte, Dennis; van Beek, Hugo L.; Syberg, Falk; Schallmey, Marcus; Tobola, Felix; Cormann, Kai U.; Schlicker, Christine; Baumann, Philipp T.; Krumbach, Karin; Sokolowsky, Sascha; Morris, Connor J.; Grünberger, Alexander; Hofmann, Eckhard; Schröder, Gunnar F.; Marienhagen, Jan
  • imprint: Springer Science and Business Media LLC, 2020
  • Published in: Nature Communications
  • Language: English
  • DOI: 10.1038/s41467-020-18400-0
  • ISSN: 2041-1723
  • Origination:
  • Footnote:
  • Description: <jats:title>Abstract</jats:title><jats:p>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<jats:italic>Corynebacterium glutamicum</jats:italic>, 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<jats:sc>l</jats:sc>-lysine insensitive LysG-based biosensor. This biosensor can be used to isolate<jats:sc>l</jats:sc>-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.</jats:p>
  • Access State: Open Access