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Characterisation of a Recombinant Patchoulol Synthase Variant for Biocatalytic Production of Terpenes

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Abstract

The patchoulol synthase (PTS) is a multi-product sesquiterpene synthases which is the central enzyme for biosynthesis of patchouli essential oil in the patchouli plant. Sesquiterpene synthases catalyse the formation of various complex carbon backbones difficult to approach by organic synthesis. Here, we report the characterisation of a recombinant patchoulol synthase complementary DNA (cDNA) variant (PTS var. 1), exhibiting significant amino acid exchanges compared to the native PTS. The product spectrum using the natural substrate E,E-farnesyl diphosphate (FDP) as well as terpenoid products resulting from conversions employing alternative substrates was analysed by GC-MS. In respect to a potential use as a biocatalyst, important enzymatic parameters such as the optimal reaction conditions, kinetic behaviour and the product selectivity were studied as well. Adjusting the reaction conditions, an increased patchoulol ratio in the recombinant essential oil was achieved. Nevertheless, the ratio remained lower than in plant-derived patchouli oil. As alternative substrates, several prenyl diposphates were accepted and converted in numerous compounds by the PTS var. 1, revealing its great biocatalytic potential.

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Acknowledgments

We would like to thank Prof. Ralf G. Berger and Dr. Ulrich Krings (both Institute of Food Chemistry, Leibniz University Hannover, Germany) for the performance of GC-MS measurements. Additionally, we would like to thank Dr. Torsten Kulke and Dr. Johannes Panten (both) for the supply with chemicals.

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Funding

This study was funded by the European Regional Development Fund (EFRE): Innovation Network “Refinement of plant resources” (ZW-8-80130940).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Hannover, Germany

    Thore Frister, Steffen Hartwig, Semra Alemdar, Katharina Schnatz, Laura Thöns, Thomas Scheper & Sascha Beutel

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  1. Thore Frister
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  2. Steffen Hartwig
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Correspondence to Sascha Beutel.

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Frister, T., Hartwig, S., Alemdar, S. et al. Characterisation of a Recombinant Patchoulol Synthase Variant for Biocatalytic Production of Terpenes. Appl Biochem Biotechnol 176, 2185–2201 (2015). https://doi.org/10.1007/s12010-015-1707-y

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