Reconstitution of Biosynthetic Machinery for the Synthesis of the Highly Elaborated Indole Diterpene Penitrem |
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| Authors: | Dr Chengwei Liu Koichi Tagami Dr Atsushi Minami Tomoyuki Matsumoto Jens Christian Frisvad Hideyuki Suzuki Jun Ishikawa Katsuya Gomi Prof Dr Hideaki Oikawa |
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| Institution: | 1. Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060‐0810 (Japan);2. Center for Eukaryotic Biotechnology, Departments of Systems Biology, Technical University of Denmark, Biocentrum‐DTU, Building 221, 2800 Kgs. Lyngby (Denmark);3. Kazusa DNA Research Institute, Kisarazu, Chiba 292‐0818 (Japan);4. Department of Bioactive Molecules, National Institute of Infectious Diseases, Shinjuku 162‐8640 (Japan);5. Graduate School of Agricultural Science, Tohoku University, Sendai 981‐8555 (Japan) |
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| Abstract: | Penitrem A is one of the most elaborated members of the fungal indole diterpenes. Two separate penitrem gene clusters were identified using genomic and RNA sequencing data, and 13 out of 17 transformations in the penitrem biosynthesis were elucidated by heterologous reconstitution of the relevant genes. These reactions involve 1) a prenylation‐initiated cationic cyclization to install the bicyclo3.2.0]heptane skeleton (PtmE), 2) a two‐step P450‐catalyzed oxidative processes forming the unique tricyclic penitrem skeleton (PtmK and PtmU), and 3) five sequential oxidative transformations (PtmKULNJ). Importantly, without conventional gene disruption, reconstitution of the biosynthetic machinery provided sufficient data to determine the pathway. It was thus demonstrated that the Aspergillus oryzae reconstitution system is a powerful method for studying the biosynthesis of complex natural products. |
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| Keywords: | biosynthesis heterologous expression indole diterpenes natural products penitrem |
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