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Biocatalytic Oxidative C[BOND]C Bond Formation Catalysed by the Berberine Bridge Enzyme: Optimal Reaction Conditions

Authors

  • Verena Resch,

    1. Department of Chemistry, Organic and Bioorganic Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria, Fax: (+43)-316-380-9840
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  • Joerg H. Schrittwieser,

    1. Department of Chemistry, Organic and Bioorganic Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria, Fax: (+43)-316-380-9840
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  • Silvia Wallner,

    1. Institute of Biochemistry, Graz University of Technology, Petersgasse 12, 8010 Graz, Austria
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  • Peter Macheroux,

    1. Institute of Biochemistry, Graz University of Technology, Petersgasse 12, 8010 Graz, Austria
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  • Wolfgang Kroutil

    Corresponding author
    1. Department of Chemistry, Organic and Bioorganic Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria, Fax: (+43)-316-380-9840
    • Department of Chemistry, Organic and Bioorganic Chemistry, University of Graz, Heinrichstrasse 28, 8010 Graz, Austria, Fax: (+43)-316-380-9840
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Abstract

Berberine bridge enzyme (BBE) catalyses the oxidative formation of an intramolecular C[BOND]C bond using (S)-reticuline as the natural substrate to form (S)-scoulerine as the product. To allow application of the enzyme on a preparative scale for the synthesis of novel optically pure berbine and isoquinoline derivatives, an organic solvent is required to solubilise the barely soluble substrates. It was shown that BBE tolerates a broad variety of organic co-solvents. Ideally the enzymatic enantioselective oxidative C[BOND]C bond formation can be performed in 70% v v−1 toluene concentration, which allowed a soluble substrate concentration of at least 20 g L−1. In addition, the enzyme works in a broad operational window concerning pH and temperature. High conversions can be reached between pH 8 and 11 and from 30 to 50 °C, respectively. The enantioselective oxidative C[BOND]C bond formation was demonstrated on a preparative scale (500 mg) in a kinetic resolution leading to optically pure products (>97% ee).

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