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Engineering and application of a biosensor with focused ligand specificity

Author

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  • Dennis Della Corte

    (Institute of Biological Information Processing, IBI-7: Structural Biochemistry, Forschungszentrum Jülich
    Brigham Young University)

  • Hugo L. Beek

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich)

  • Falk Syberg

    (Protein Crystallography, Biophysics, Ruhr University Bochum)

  • Marcus Schallmey

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich)

  • Felix Tobola

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich)

  • Kai U. Cormann

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich)

  • Christine Schlicker

    (Protein Crystallography, Biophysics, Ruhr University Bochum)

  • Philipp T. Baumann

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich)

  • Karin Krumbach

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich)

  • Sascha Sokolowsky

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich)

  • Connor J. Morris

    (Brigham Young University)

  • Alexander Grünberger

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich
    Multiscale Bioengineering, Bielefeld University)

  • Eckhard Hofmann

    (Protein Crystallography, Biophysics, Ruhr University Bochum)

  • Gunnar F. Schröder

    (Institute of Biological Information Processing, IBI-7: Structural Biochemistry, Forschungszentrum Jülich
    Heinrich-Heine University
    Jülich Centre for Structural Biology (JuStruct), Forschungszentrum Jülich)

  • Jan Marienhagen

    (Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich
    Institute of Biotechnology, RWTH Aachen University)

Abstract

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.

Suggested Citation

  • Dennis Della Corte & Hugo L. Beek & Falk Syberg & Marcus Schallmey & Felix Tobola & Kai U. Cormann & Christine Schlicker & Philipp T. Baumann & Karin Krumbach & Sascha Sokolowsky & Connor J. Morris & , 2020. "Engineering and application of a biosensor with focused ligand specificity," Nature Communications, Nature, vol. 11(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-18400-0
    DOI: 10.1038/s41467-020-18400-0
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    Cited by:

    1. Kumar, Premashis & Banerjee, Kinshuk & Gangopadhyay, Gautam, 2022. "Interplay of energy, dissipation, and error in kinetic proofreading: Control via concentration and binding energy," Physica A: Statistical Mechanics and its Applications, Elsevier, vol. 603(C).

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