Development of versatile synthesis for fungal natural products by reconstitution of biosynthetic machinery

• Project/Area Number: 15H01835
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: Hokkaido University
• Principal Investigator: Oikawa Hideaki 北海道大学, 理学研究院, 教授 (00185175)
• Co-Investigator(Kenkyū-buntansha): 南 篤志 北海道大学, 理学研究院, 准教授 (40507191)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥40,820,000 (Direct Cost: ¥31,400,000、Indirect Cost: ¥9,420,000); Fiscal Year 2017: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2016: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2015: ¥27,040,000 (Direct Cost: ¥20,800,000、Indirect Cost: ¥6,240,000)
• Keywords: 生合成 / 天然物 / テルペン / ペプチド / 麹菌

Outline of Final Research Achievements

Using Aspergillus oryzae expression system, we have succeeded elucidation of enzymatic mechanism of several important natural products such as C20/C25-terpenes, ribosomal peptide and mushroom antibiotics. Especially, we proved the expression system is reliable for production of known and novel natural products (terpenoids) using uncharacterized biosynthetic gene clusters.

Research Products

• [Int'l Joint Research] Massey University(New Zealand)
• [Int'l Joint Research] Technical University of Denmark(Denmark)
• [Journal Article] Subcellular localization of aphidicolin biosynthetic enzymes heterologously expressed in Aspergillus oryzae (2018)
  • Author(s): A. Ban, M. Tanaka, R. Fujii, A. Minami, H. Oikawa, T. Shintani, K. Gomi
  • Journal Title: Bioscience Biotechnology and Biochemistry Volume: 82 Issue: 1 Pages: 139-147
  • DOI: 10.1080/09168451.2017.1399789
  • Peer Reviewed
• [Journal Article] Total Biosynthesis of Antiangiogenic Agent (-)-Terpestacin by Artificial Reconstitution of the Biosynthetic Machinery in Aspergillus oryzae (2018)
  • Author(s): Narita Koji、Minami Atsushi、Ozaki Taro、Liu Chengwei、Kodama Motoichiro、Oikawa Hideaki
  • Journal Title: The Journal of Organic Chemistry Volume: 83 Issue: 13 Pages: 7042-7048
  • DOI: 10.1021/acs.joc.7b03220
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Theoretical Study of Sesterfisherol Biosynthesis: Computational Prediction of Key Amino Acid Residue in Terpene Synthase (2018)
  • Author(s): Sato Hajime、Narita Koji、Minami Atsushi、Yamazaki Mami、Wang Chao、Suemune Hironori、Nagano Shingo、Tomita Takeo、Oikawa Hideaki、Uchiyama Masanobu
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 2473-2473
  • DOI: 10.1038/s41598-018-20916-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of novel sesterterpenes by genome mining of phytopathogenic fungi Phoma and Colletotrichum sp. (2018)
  • Author(s): Gao Lei、Narita Koji、Ozaki Taro、Kumakura Naoyoshi、Gan Pamela、Minami Atsushi、Liu Chengwei、Lei Xiaoguang、Shirasu Ken、Oikawa Hideaki
  • Journal Title: Tetrahedron Letters Volume: 59 Issue: 12 Pages: 1136-1139
  • DOI: 10.1016/j.tetlet.2018.02.022
  • NAID: 120006814278
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Enzymatic formation of a skipped methyl-substituted octaprenyl side chain of longestin (KS-505a): Involvement of homo-IPP as a common extender unit (2018)
  • Author(s): Ozaki Taro、Shinde Sandip S.、Gao Lei、Okuizumi Ryo、Liu Chengwei、Ogasawara Yasushi、Lei Xiaoguang、Dairi Tohru、Minami Atsushi、Oikawa Hideaki
  • Journal Title: Angewandte Chemie International Edition Volume: 57 Issue: 22 Pages: 6629-6632
  • DOI: 10.1002/anie.201802116
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Stepwise cyclopropanation on the polycyclopropanated polyketide formation in jawsamycin biosynthesis (2017)
  • Author(s): Hiratsuka, T.; Suzuki, H.; Minami, A.; Oikawa, H.
  • Journal Title: Organic and Biomolecular Chemistry Volume: 15 Issue: 5 Pages: 1076-1079
  • DOI: 10.1039/c6ob02675c
  • NAID: 120006383812
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Focused genome mining of structurally related sesterterpenes: enzymatic formation of enantiomeric and diastereomeric products (2017)
  • Author(s): Narita, K., Sato, H., Minami, A., Kudo, K., Gao, L., Liu, C., Ozaki, T., Kodama, M.., Lei, X., Taniguchi, T., Monde, K., Yamazaki, M., Uchiyama, M., Oikawa, H.
  • Journal Title: Organic letters Volume: 19 Issue: 24 Pages: 6696-6699
  • DOI: 10.1021/acs.orglett.7b03418
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Biosynthetic Machinery of Diterpene Pleuromutilin Isolated from Basidiomycete Fungi (2017)
  • Author(s): Yamane Momoka、Minami Atsushi、Liu Chengwei、Ozaki Taro、Takeuchi Ichiro、Tsukagoshi Tae、Tokiwano Tetsuo、Gomi Katsuya、Oikawa Hideaki
  • Journal Title: ChemBioChem Volume: 18 Issue: 23 Pages: 2317-2322
  • DOI: 10.1002/cbic.201700434
  • NAID: 120006541883
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Unveiling the Biosynthetic Pathway of the Ribosomally Synthesized and Post-translationally Modified Peptide Ustiloxin B in Filamentous Fungi. (2016)
  • Author(s): Y. Ye, A. Minami, Y. Igarashi, M. Izumikawa, M. Umemura, N. Nagano, M. Machida, T. Kawahara, K. Shin-ya, K. Gomi, H. Oikawa
  • Journal Title: Angewandte Chemie-International Edition Volume: 55 Issue: 28 Pages: 8072-8075
  • DOI: 10.1002/anie.201602611
  • NAID: 120006318732
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Multiple oxidative modifications in the ophiobolin biosynthesis: P450 oxidations found in the genome mining (2016)
  • Author(s): K. Narita, R. Chiba, A. Minami, M. Kodama, I. Fujii, K. Gomi and H. Oikawa
  • Journal Title: Org. Lett. Volume: 18 Issue: 9 Pages: 1980-1983
  • DOI: 10.1021/acs.orglett.6b00552
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Biosynthesis of shearinine: Diversification of a tandem prenyl moiety of fungal indole diterpenes (2016)
  • Author(s): Liu, C.; Minami, A.; Dairi, T.; Gomi, K.; Scott, B.; Oikawa, H.
  • Journal Title: Organic Letters Volume: 18 Issue: 19 Pages: 5026-5029
  • DOI: 10.1021/acs.orglett.6b02482
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Genome mining approach for harnessing the cryptic gene cluster in Alternaria solani: production of PKS-NRPS hybrid metabolite, didymellamide B. (2016)
  • Author(s): T. Ugai, A. Minami, K. Gomi, H. Oikawa
  • Journal Title: Tetrahedron Letters Volume: 57 Issue: 25 Pages: 2793-2796
  • DOI: 10.1016/j.tetlet.2016.05.043
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Reconstitution of biosynthetic machinery of fungal polyketides: unexpected oxidations of biosynthetic intermediates by expression host (2016)
  • Author(s): R. Fujii, T. Ugai, H. Ichinose, M. Hatakeyama, T. Kosaki, K. Gomi, I. Fujii, A. Minami, and H. Oikawa
  • Journal Title: Biosci. Biotechnol. Biochem. Volume: 80 Issue: 3 Pages: 426-431
  • DOI: 10.1080/09168451.2015.1104234
  • NAID: 120005981344
  • Peer Reviewed
• [Journal Article] Catalytic asymmetric synthesis of the common amino acid component in the biosynthesis of tetrahydroisoquinoline alkaloids. (2016)
  • Author(s): R. Tanifuji, H. Oguri, K. Koketsu, Y. Yoshinaga, A. Minami, H. Oikawa
  • Journal Title: Tetrahedron Letters Volume: 57 Issue: 5 Pages: 623-626
  • DOI: 10.1016/j.tetlet.2015.12.110
  • Peer Reviewed
• [Journal Article] Total Biosynthesis of Fungal Indole Diterpenes Using Cell Factories. (2016)
  • Author(s): A. Minami, C. W. Liu, H. Oikawa
  • Journal Title: Heterocycles Volume: 92 Issue: 3 Pages: 397-421
  • DOI: 10.3987/rev-15-830
  • NAID: 40020766375
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Recent advances of Diels-Alderases involved in natural product biosynthesis. (2016)
  • Author(s): A. Minami, H. Oikawa
  • Journal Title: Journal of Antibiotics Volume: 69 Issue: 7 Pages: 500-506
  • DOI: 10.1038/ja.2016.67
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Minimum Information About a Biosynthetic Gene Cluster (2015)
  • Author(s): M. H. Medema, H. Oikawa 他
  • Journal Title: Nat. Chem. Biol. Volume: 11 Issue: 9 Pages: 625-631
  • DOI: 10.1038/nchembio.1890
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Biosynthetic Study on Antihypercholesterolemic Agent Phomoidride: General Biogenesis of Fungal Dimeric Anhydrides (2015)
  • Author(s): R. Fujii, Y. Matsu, A. Minami, S. Nagamine, I. Takeuchi, K. Gomi, and H. Oikawa
  • Journal Title: Org. Lett. Volume: 17 Issue: 22 Pages: 5658-5661
  • DOI: 10.1021/acs.orglett.5b02934
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Genome Mining for Sesterterpenes Using Bifunctional Terpene Synthases Reveals a Unified Intermediate of Di/Sesterterpenes (2015)
  • Author(s): Y. Ye, A. Minami, A. Mandi, C. Liu, T. Taniguchi, T. Kuzuyama, K. Monde, K. Gomi, and H. Oikawa
  • Journal Title: J. Am. Chem. Soc. Volume: 137 Issue: 36 Pages: 11846-11853
  • DOI: 10.1021/jacs.5b08319
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Reconstitution of biosynthetic machinery for the synthesis of the highly elaborated indole diterpene Penitrem. (2015)
  • Author(s): Liu, C., Tagami, K., Minami, A., Matsumoto, T., Frisvad, J.C., Suzuki, H., Ishikawa, J., Gomi, K., and Oikawa, H.
  • Journal Title: Angew. Chem. Int. Ed. Volume: 54 Issue: 19 Pages: 5748-5752
  • DOI: 10.1002/anie.201501072
  • NAID: 130007492191
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Heterologous expression of highly reducing polyketide synthase involved in betaenone biosynthesis (2015)
  • Author(s): T. Ugai, A. Minami, R. Fujii, M. Tanaka, K. Gomi, H. Oikawa
  • Journal Title: Chem. Commun Volume: 51 Issue: 10 Pages: 1878-1878
  • DOI: 10.1039/c4cc09512j
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Nature’s strategy for synthesis of biologically active small molecule (2018)
  • Author(s): Hideaki Oikawa
  • Organizer: Students Seminar of NIMS Joint Graduate Program
  • Invited
• [Presentation] Biosynthetic mechanism of fungal ribosomal peptides RiPPS (2018)
  • Author(s): Taro Ozaki, Chengwei Liu, Atsushi Minami, Hideaki Oikawa
  • Organizer: 農芸化学会2018年度大会シンポジウム“多様なペプチド化合物を創出する微生物が備えた生合成戦略”
  • Invited
• [Presentation] 生物が選択した天然物合成戦略：糸状菌天然物を中心に (2017)
  • Author(s): 及川英秋
  • Organizer: 日本化学会 第97春季年会特別企画新領域研究グループシンポジウム
  • Place of Presentation: 慶應義塾大学（神奈川県・横浜市）
  • Year and Date: 2017-03-16
  • Invited
• [Presentation] ゲノム情報を用いた有用生物活性を持つ低分子化合物の汎用的酵素合成法の開発 (2017)
  • Author(s): 及川 英秋
  • Organizer: 北大－ＮＩＭＳジョイントシンポジウム
  • Invited
• [Presentation] Total biosynthetic study of fungal metabolites (2017)
  • Author(s): Hideaki Oikawa
  • Organizer: 9th US-Japan Seminar on the Biosynthesis of Natural Products
  • Int'l Joint Research / Invited
• [Presentation] 骨格構造が類似した天然物の集中的生合成法の開発 (2017)
  • Author(s): 及川 英秋
  • Organizer: 日本ケミカルバイオロジー学会第12回年会特別講演“有機合成化学を起点とするものづくり戦略”
  • Int'l Joint Research / Invited
• [Presentation] Construction of Focused Library of Structurally Related Fungal Metabolites Using Genome Mining (2017)
  • Author(s): 及川 英秋
  • Organizer: 1st China-Japan Joint Symposium on Natural Product Biosynthesis
  • Int'l Joint Research / Invited
• [Presentation] 生合成マシナリー再構築による天然物合成の新展開 (2017)
  • Author(s): 及川 英秋
  • Organizer: 東京農工大学大学院特別講演会
  • Invited
• [Presentation] Construction of focused library of structurally related fungal metabolites (2017)
  • Author(s): Atsushi Minami, Chengwei Liu, Taro Ozaki, Hideaki Oikawa
  • Organizer: The Second A3 Roundtable Meeting on Chemical Probe Research Hub
  • Int'l Joint Research / Invited
• [Presentation] 天然物生合成マシナリーから生合成リデザインへ：麹菌による驚異的異種発現 (2016)
  • Author(s): 及川英秋
  • Organizer: 生物合成系の再設計による複雑骨格機能分子の革新的創成科学 新学術領域「生合成リデザイン」キックオフシンポジウム
  • Place of Presentation: 東京大学薬学系総合研究棟(東京都・文京区）
  • Year and Date: 2016-09-10
  • Invited
• [Presentation] Recent Topics for Total Biosynthesis of Fungal Natural Products (2016)
  • Author(s): Hideaki Oikawa
  • Organizer: Peking University & Hokkaido University Joint Seminar on Organic Chemistry and Chemical Biology
  • Place of Presentation: Peking University, Beijing, China
  • Year and Date: 2016-05-26
  • Int'l Joint Research / Invited
• [Presentation] Total Biosynthesis of Microbial Natural Products (2016)
  • Author(s): H. Oikawa
  • Organizer: The 8th Takeda Science Foundation Symposium
  • Place of Presentation: Center for Learning and Innovation (Suita, Osaka)
  • Year and Date: 2016-01-21
  • Int'l Joint Research / Invited
• [Presentation] Novel enzymatic reaction mechanism found in biosynthesis of natural products (2015)
  • Author(s): H. Oikawa
  • Organizer: Bioorganic Reaction Mechanisms, Pacifichem2015
  • Place of Presentation: Honolulu (USA)
  • Year and Date: 2015-12-19
  • Int'l Joint Research / Invited
• [Presentation] Total biosynthetic study of bioactive fungal natural products: Pathway elucidation and genome mining (2015)
  • Author(s): H. Oikawa
  • Organizer: Biosynthesis of Natural Products, Pacifichem2015
  • Place of Presentation: Honolulu (USA)
  • Year and Date: 2015-12-18
  • Int'l Joint Research / Invited
• [Presentation] Study on the enzymatic construction of polycyclopropane skeleton in jawsamycin biosynthesis (2015)
  • Author(s): Atsushi Minami , Tomoshige Hiratsuka , Hideaki Oikawa
  • Organizer: Pacifichem2015
  • Place of Presentation: Honolulu (USA)
  • Year and Date: 2015-12-18
  • Int'l Joint Research / Invited
• [Presentation] Reconstitution of biosynthetic machinery for highly elaborated indole diterpene penitrem in Aspergillus oryzea (2015)
  • Author(s): Liu Chengwei , Koichi Tagami , Atsushi Minami , Tomoyuki Matsumoto , Hideaki Oikawa
  • Organizer: Pacifichem2015
  • Place of Presentation: Honolulu (USA)
  • Year and Date: 2015-12-18
  • Int'l Joint Research
• [Presentation] Genome mining of sesterterpenes by bifunctional terpene synthase: Unified biogenesis of sesterterpenes (2015)
  • Author(s): Ying Ye, Atsushi Minami, Katsuya Gomi, Hideaki Oikawa
  • Organizer: 第25回イソプレノイド研究会
  • Place of Presentation: 東北大学片平さくらホール（宮城県・仙台市）
  • Year and Date: 2015-09-14
• [Presentation] 麹菌異種発現系を利用したペニトレム生合成マシナリーの解明 (2015)
  • Author(s): 南 篤志、劉 成偉、田上 紘一、松本 知之、Jens Christian Frisvad, 鈴木 秀幸、石川 淳、五味 勝也、及川 英秋
  • Organizer: 第57回天然有機化合物討論会
  • Place of Presentation: 神奈川県民ホール（神奈川県・横浜市）
  • Year and Date: 2015-09-11
• [Presentation] 生合成マシナリー再構築法による 糸状菌天然物の全生合成 (2015)
  • Author(s): 及川 英秋
  • Organizer: 天然有機化合物の生合成研究―新潟ミニシンポジウム―
  • Place of Presentation: トキメイト（新潟県、新潟市）
  • Year and Date: 2015-05-22
  • Invited
• [Book] 酵母菌・麹菌・乳酸菌の産業応用展開 (2018)
  • Author(s): 及川英秋,南篤志,劉誠偉,尾﨑太郎,他69名 監修：五味勝也,阿部敬悦
  • Total Pages: 264
  • Publisher: シーエムシー出版
  • ISBN: 9784781313177
• [Remarks] 有機反応論研究室
  • URL: https://wwwchem.sci.hokudai.ac.jp/~yuhan/index.html