Engineering and application of a biosensor with focused ligand specificity

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, Alexander Grünberger, Eckhard Hofmann, Gunnar F. Schröder and Jan Marienhagen ()
Additional contact information
Dennis Della Corte: Institute of Biological Information Processing, IBI-7: Structural Biochemistry, Forschungszentrum Jülich
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
Eckhard Hofmann: Protein Crystallography, Biophysics, Ruhr University Bochum
Gunnar F. Schröder: Institute of Biological Information Processing, IBI-7: Structural Biochemistry, Forschungszentrum Jülich
Jan Marienhagen: Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich

Nature Communications, 2020, vol. 11, issue 1, 1-11

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-18400-0

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