A Combination of In Vivo Selection and Cell Sorting for the Identification of Enantioselective Biocatalysts

Authors

  • Dr. Elena Fernández-Álvaro,

    1. Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
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  • Dr. Radka Snajdrova,

    1. Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
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  • Dr. Helge Jochens,

    1. Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
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  • Dipl.-Biol. Timo Davids,

    1. Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
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  • Dr. Dominique Böttcher,

    Corresponding author
    1. Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
    • Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
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  • Prof. Dr. Uwe T. Bornscheuer

    Corresponding author
    1. Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
    • Institute of Biochemistry, Dept. of Biotechnology & Enzyme Catalysis, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany)
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  • We thank the Deutsche Bundesstiftung Umwelt (grant AZ 13198) and the DFG (grant Bo1862/4-1) for financial support.

Abstract

original image

The “carrot and stick” principle could be combined with cell sorting to enable the selection of enantioselective esterase variants from a mutant library. Hence, the enormous diversity generated in directed evolution experiments is now easily accessible by this high-throughput system. In line with the principle, the hydrolysis of 1 glycerin supports cell growth, whereas the hydrolysis of 2 leads to cell death.

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