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Identification, Characterization, and Application of Three Enoate Reductases from Pseudomonas putida in In Vitro Enzyme Cascade Reactions

Authors

  • Christin Peters,

    1. Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, 17487 Greifswald (Germany), Fax: (+49) 3834-86-794367
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    • These authors contributed equally to this work.

  • Regina Kölzsch,

    1. Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, 17487 Greifswald (Germany), Fax: (+49) 3834-86-794367
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    • These authors contributed equally to this work.

  • Dr. Maria Kadow,

    1. Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, 17487 Greifswald (Germany), Fax: (+49) 3834-86-794367
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  • Lilly Skalden,

    1. Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, 17487 Greifswald (Germany), Fax: (+49) 3834-86-794367
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  • Dr. Florian Rudroff,

    1. Institute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-OC, 1060 Vienna (Austria)
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  • Prof. Dr. Marko D. Mihovilovic,

    1. Institute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-OC, 1060 Vienna (Austria)
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  • Prof. Dr. Uwe T. Bornscheuer

    Corresponding author
    1. Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, 17487 Greifswald (Germany), Fax: (+49) 3834-86-794367
    • Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, 17487 Greifswald (Germany), Fax: (+49) 3834-86-794367

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Abstract

Enoate reductases are versatile enzymes for the enantio- and regioselective addition of hydrogen to double bonds. We identified three EREDs (XenA, XenB, NemA) from Pseudomonas putida ATCC 17453 through a sequence motif search. In addition to cloning, functional expression, and biochemical characterization of these enzymes, the enoate reductases were also applied in enzyme cascade reactions in combination with a Baeyer–Villiger monooxygenase and an alcohol dehydrogenase to produce lactones.

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