Engineering and application of a biosensor with focused ligand specificity
Nature Communications, vol. 11, pp. 4851
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.
Authors 15
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Brigham Young University · Forschungszentrum Jülich
Affiliation as printed
Department of Physics & Astronomy, Brigham Young University, Provo, UT, USA
Institute of Biological Information Processing, IBI-7: Structural Biochemistry, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Protein Crystallography, Biophysics, Ruhr University Bochum, D-44780, Bochum, Germany
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Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Protein Crystallography, Biophysics, Ruhr University Bochum, D-44780, Bochum, Germany
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Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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Affiliation as printed
Department of Physics & Astronomy, Brigham Young University, Provo, UT, USA
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Forschungszentrum Jülich · Bielefeld University
Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
Multiscale Bioengineering, Bielefeld University, D-33615, Bielefeld, Germany
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Affiliation as printed
Protein Crystallography, Biophysics, Ruhr University Bochum, D-44780, Bochum, Germany
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Forschungszentrum Jülich · Heinrich Heine University Düsseldorf
Affiliation as printed
Department of Physics, Heinrich-Heine University, D-40225, Düsseldorf, Germany
Institute of Biological Information Processing, IBI-7: Structural Biochemistry, Forschungszentrum Jülich, D-52425, Jülich, Germany
Jülich Centre for Structural Biology (JuStruct), Forschungszentrum Jülich, D-52425, Jülich, Germany
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Forschungszentrum Jülich · RWTH Aachen University
Affiliation as printed
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany. j.marienhagen@fz-juelich.de
Institute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074, Aachen, Germany. j.marienhagen@fz-juelich.de
Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany
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