Engineered biosynthesis of new natural products for innovative drug discovery

• Project Area: Creation of Complex Functional Molecules by Rational Redesign of Biosynthetic Machineries
• Project/Area Number: 16H06449
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: University of Shizuoka
• Principal Investigator: 渡辺 賢二 静岡県立大学, 薬学部, 教授 (50360938)
• Co-Investigator(Kenkyū-buntansha): 植木 尚子 岡山大学, 資源植物科学研究所, 准教授 (50622023)
• Project Period (FY): 2016-06-30 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥72,020,000 (Direct Cost: ¥55,400,000、Indirect Cost: ¥16,620,000); Fiscal Year 2020: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000); Fiscal Year 2019: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000); Fiscal Year 2018: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000); Fiscal Year 2017: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000); Fiscal Year 2016: ¥15,860,000 (Direct Cost: ¥12,200,000、Indirect Cost: ¥3,660,000)
• Keywords: 天然物 / 生合成 / リデザイン / 遺伝子クラスター / 活性物質 / 担子菌 / 生理活性物質 / 抗生物質 / 生合成リデザイン / テルペノイド / 天然物化学 / キノコ / 酵素 / 遺伝子 / 微量天然物 / 異種発現 / 生合 / 反応機構 / 糸状菌 / Aspoquinolone / ゲノム情報 / 物質生産

Outline of Annual Research Achievements

C. cinereaは、CPFを産生・分泌することで細胞外の鉄を効率的に細胞内に取り込んでいると考えられる。そして取り込まれた鉄が菌糸成長の促進・子実体形成に重要な働きを有すると本研究にて示された。CPF生合成遺伝子クラスター(CPFクラスター)はゲノムデータベース上に登録された担子菌類のうち200種類を超える担子菌種で高度に保存されており、なかには我々が普段口にするようなブナシメジやマイタケのような菌種も含まれている。実際に我々はCPFクラスターを有する担子菌種や市販の食用キノコにおいてCPFの生産を確認している。以上は、CPFが多くの担子菌種の菌糸成長・子実体形成に関与する共通した普遍的分子であることを示唆している。通常、二次代謝産物は、それぞれの生産種において独自性の高い代謝経路で生産され、自身の生育に直接的に関与しない物質として定義されている。一方で、CPF合成不全株KOCclaeAKOcpf1が致死の表現型を示さないことは、CPFが一次代謝産物とも異なることを意味する。このようにCPFは天然物のなかでも独自に位置づけされる生理機能性分子であり、担子菌が如何にしてCPF分子を生合成・利用できるように進化・適応したのか興味がもたれる。現在は、CPFを用いてマツタケなどの人工栽培が難しい担子菌種の培養を可能にするのではないかと期待し研究を行っている。また、一般的に子嚢菌由来laeAは二次代謝産物生産を正に制御するのに対し、予想外にも担子菌由来CclaeAはCPFの生産を負に制御していた。本研究はLaeAタンパク質の二次代謝制御における新たな機能を世に提示するものであり、本機能の解明により生合成遺伝子の休眠・覚醒化の理解、ひいては効率的な新規天然物獲得法の樹立へとつながると期待される。

Research Progress Status

令和2年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和2年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] UCLA(米国)
• [Int'l Joint Research] University of California Los Angeles(米国)
• [Int'l Joint Research] Leibniz Institute(ドイツ)
• [Int'l Joint Research] カリフォルニア大ロサンゼルス校(米国)
• [Int'l Joint Research] University of California, Los Angeles(USA)
• [Int'l Joint Research]
• [Journal Article] Catalytic mechanism and endo-to-exo selectivity reversion of an octalin-forming natural Diels?Alderase (2021)
  • Author(s): Sato Michio、Kishimoto Shinji、Yokoyama Mamoru、Jamieson Cooper S.、Narita Kazuto、Maeda Naoya、Hara Kodai、Hashimoto Hiroshi、Tsunematsu Yuta、Houk Kendall N.、Tang Yi、Watanabe Kenji
  • Journal Title: Nature Catalysis Volume: 4 Pages: 223-232
  • DOI: 10.1038/s41929-021-00577-2
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Concise Biosynthesis of Phenylfuropyridones in Fungi (2020)
  • Author(s): Zhang Zhuan、Qiao Tianzhang、Watanabe Kenji、Tang Yi
  • Journal Title: Angewandte Chemie International Edition Volume: 59 Pages: 19889-19893
  • DOI: 10.1002/anie.202008321
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Biosynthesis of the Immunosuppressant (?)-FR901483 (2020)
  • Author(s): Zhang Zhuan、Tamura Yui、Tang Mancheng、Qiao Tianzhang、Sato Michio、Otsu Yoshihiro、Sasamura Satoshi、Taniguchi Masatoshi、Watanabe Kenji、Tang Yi
  • Journal Title: Journal of the American Chemical Society Volume: 143 Pages: 132-136
  • DOI: 10.1021/jacs.0c12352
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Specialized Flavoprotein Promotes Sulfur Migration and Spiroaminal Formation in Aspirochlorine Biosynthesis (2020)
  • Author(s): Tsunematsu Yuta、Maeda Naoya、Sato Michio、Hara Kodai、Hashimoto Hiroshi、Watanabe Kenji、Hertweck Christian
  • Journal Title: Journal of the American Chemical Society Volume: 143 Pages: 206-213
  • DOI: 10.1021/jacs.0c08879
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Uncovering hidden sesquiterpene biosynthetic pathway through expression boost area-mediated productivity enhancement in basidiomycete (2020)
  • Author(s): Asai Shihori、Tsunematsu Yuta、Masuya Takahiro、Otaka Junnosuke、Osada Hiroyuki、Watanabe Kenji
  • Journal Title: J Antibiot Volume: 73 Pages: 721-728
  • DOI: 10.1038/s41429-020-0355-9
  • Peer Reviewed
• [Journal Article] Bisindole (2020)
  • Author(s): Sato, M., Kishimoto, S., Noguchi, H., Watanabe, K.
  • Journal Title: Comprehensive Natural Products III: Chemistry and Biology, Wiley Volume: 4 Pages: 1-1
  • Peer Reviewed
• [Journal Article] Oxidative modification enzymes in polyketide biosynthetic pathways (2020)
  • Author(s): Tsunematsu, Y., Hirayama, Y., Masuya, T., Watanabe, K.
  • Journal Title: Comprehensive Natural Products III: Chemistry and Biology, Wiley Volume: 4 Pages: 30-30
  • Peer Reviewed
• [Journal Article] A new class of dimeric product isolated from the fungus Chaetomium globosum: evaluation of chemical structure and biological activity (2020)
  • Author(s): Sarmales-Murga Christopher、Akaoka Fumito、Sato Michio、Takanishi Jun、Mino Takashi、Miyoshi Noriyuki、Watanabe Kenji
  • Journal Title: The Journal of Antibiotics Volume: 73 Pages: 320-323
  • DOI: 10.1038/s41429-020-0281-x
  • Peer Reviewed
• [Journal Article] Genomic Mushroom Hunting Decrypts Coprinoferrin, A Siderophore Secondary Metabolite Vital to Fungal Cell Development (2019)
  • Author(s): Tsunematsu Yuta、Takanishi Jun、Asai Shihori、Masuya Takahiro、Nakazawa Takehito、Watanabe Kenji
  • Journal Title: Organic Letters Volume: 21 Pages: 7582-7586
  • DOI: 10.1021/acs.orglett.9b02861
  • Peer Reviewed
• [Journal Article] Functional and Structural Analyses of trans C-Methyltransferase in Fungal Polyketide Biosynthesis (2019)
  • Author(s): Kishimoto Shinji、Tsunematsu Yuta、Matsushita Takuma、Hara Kodai、Hashimoto Hiroshi、Tang Yi、Watanabe Kenji
  • Journal Title: Biochemistry, Volume: 58 Pages: 3933-3937
  • DOI: 10.1021/acs.biochem.9b00702
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Biosynthesis of lagopodins in mushroom involves a complex network of oxidation reactions (2019)
  • Author(s): Masuya Takahiro、Tsunematsu Yuta、Hirayama Yuichiro、Sato Michio、Noguchi Hiroshi、Nakazawa Takehito、Watanabe Kenji
  • Journal Title: Org Biomol Chem Volume: 17 Pages: 234-239
  • DOI: 10.1039/c8ob02814a
  • Peer Reviewed
• [Journal Article] Enzymatic one-step ring contraction for quinolone biosynthesis (2018)
  • Author(s): Kishimoto Shinji、Hara Kodai、Hashimoto Hiroshi、Hirayama Yuichiro、Champagne Pier Alexandre、Houk Kendall N.、Tang Yi、Watanabe Kenji
  • Journal Title: Nature Commun Volume: 9 Pages: 1-1
  • DOI: 10.1038/s41467-018-05221-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genome Mining and Assembly-Line Biosynthesis of the UCS1025A Pyrrolizidinone Family of Fungal Alkaloids (2018)
  • Author(s): Li Li、Tang Man-Cheng、Tang Shoubin、Gao Shushan、Soliman Sameh、Hang Leibniz、Xu Wei、Ye Tao、Watanabe Kenji、Tang Yi
  • Journal Title: Journal of the American Chemical Society Volume: 140 Pages: 2067-2071
  • DOI: 10.1021/jacs.8b00056
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Enzymatic Amide Tailoring Promotes Retro-Aldol Amino Acid Conversion To Form the Antifungal Agent Aspirochlorine (2018)
  • Author(s): Tsunematsu Yuta、Maeda Naoya、Yokoyama Mamoru、Chankhamjon Pranatchareeya、Watanabe Kenji、Scherlach Kirstin、Hertweck Christian
  • Journal Title: Angewandte Chemie International Edition Volume: 57 Pages: 14051-14054
  • DOI: 10.1002/anie.201806740
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Polyketide synthase-nonribosomal peptide synthetase hybrid enzymes of fungi (2018)
  • Author(s): Kishimoto, S., Hirayama, Y., Watanabe, K.
  • Journal Title: The Mycota XV: Physiology and Genetics Volume: 368 Pages: 1-1
  • Peer Reviewed
• [Journal Article] Integration of Chemical, Genetic, and Bioinformatic Approaches Delineates Fungal Polyketide-Peptide Hybrid Biosynthesis (2017)
  • Author(s): Yokoyama M, Hirayama Y, Yamamoto T, Kishimoto S, Tsunematsu Y, Watanabe K
  • Journal Title: Org Lett Volume: 19 Pages: 2002-2005
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] SAM-dependent enzyme-catalysed pericyclic reactions in natural product biosynthesis (2017)
  • Author(s): 1.Ohashi, M., Liu, F., Hai, Y., Chen, M., Tang, M.-C., Yang, Z., Sato, M., Watanabe, K., Houk, K. N., Tang, Y.
  • Journal Title: Nature Volume: 549 Pages: 502-506
  • DOI: 10.1038/nature23882
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Oxidative Cyclization in Natural Product Biosynthesis (2017)
  • Author(s): Tang Man-Cheng、Zou Yi、Watanabe Kenji、Walsh Christopher T.、Tang Yi
  • Journal Title: Chemical Reviews Volume: 117 Pages: 5226-5333
  • DOI: 10.1021/acs.chemrev.6b00478
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Elucidation of biosynthetic pathways of natural products. (2017)
  • Author(s): Kishimoto, S., Tsunematsu, Y., Sato, M., Watanabe, K.
  • Journal Title: The Chemical Record Volume: 印刷中 Pages: 1095-1108
  • DOI: 10.1002/tcr.201700015
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Integration of chemical, genetic and bioinformatic approaches delineates fungal polyketide-peptide hybrid biosynthesis. (2017)
  • Author(s): Yokoyama, M., Hirayama, Y., Yamamoto, T., Kishimoto, S., Tsunematsu, Y., Watanabe, K.
  • Journal Title: Org. Lett. Volume: 印刷中
  • DOI: 10.1021/acs.orglett.7b00559
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Biochemical characterization of an eukaryotic decalin-forming Diels-Alderase. (2016)
  • Author(s): Li, L., Yu, P., Tang, M. C., Zou, Y., Gao, S.-S., Hung, Y.-S., Zhao, M., Watanabe, K., Houk, K., Tang, Y.
  • Journal Title: J. Am. Chem. Soc. Volume: 138 Pages: 15837-15840
  • DOI: 10.1021/jacs.6b10452
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Regioselective dichlorination of a non-activated aliphatic carbon and phenol bismethylation by a multifunctional fungal flavoenzyme. (2016)
  • Author(s): Chankhamjon, P., Tsunematsu, Y., Ishida-Ito, M., Sasa, Y., Meyer, F., Boettger-Schmidt, D., Urbansky, B., Menzel, K. D., Scherlach, K., Watanabe, K., Hertweck, C.
  • Journal Title: Angew. Chem. Int. Ed. Volume: 55 Pages: 11955-11959
  • DOI: 10.1002/anie.201604516
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Oxidative trans to cis Isomerization of Olefins in Polyketide Biosynthesis (2016)
  • Author(s): Tsuyoshi Yamamoto, Yuta Tsunematsu, Kodai Hara, Tomohiro Suzuki, Shinji Kishimoto, Hirokazu Kawaguchi, Hiroshi Noguchi, Hiroshi Hashimoto, Yi Tang, Kinya Hotta, & Kenji Watanabe
  • Journal Title: Angewandte Chemie International Edition Volume: 印刷中 Pages: 6207-6210
  • DOI: 10.1002/anie.201600940
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Diels-Alderaseのジアステレオ選択性および生成物阻害回避メカニズムの解明 (2020)
  • Author(s): 渡辺 賢二
  • 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、第8回公開シンポジウム
  • Int'l Joint Research / Invited
• [Presentation] Diastereoselectivity and product inhibition evasion by Diels-Alderase (2020)
  • Author(s): 渡辺 賢二
  • Organizer: 1st International Conference on Natural Product Chemistry in Taiwan
  • Int'l Joint Research / Invited
• [Presentation] Diastereoselectivity of an octalin-forming Diels-Alderase (2020)
  • Author(s): 渡辺 賢二
  • Organizer: The 10th Korea-Japan Chemical Biology Symposium and 30th Meeting for New Drug Discovery
  • Int'l Joint Research / Invited
• [Presentation] Crystallographic characterization of Diels-Alderase binding site and mutational alteration of stereoselectivity (2020)
  • Author(s): 渡辺 賢二
  • Organizer: Let’s leap! Challenges in Organic Chemistry
  • Int'l Joint Research / Invited
• [Presentation] Crystallographic characterization of a [4+2]-cyclase binding site and mutational alteration of stereoselectivity (2020)
  • Author(s): 渡辺 賢二
  • Organizer: The 2nd International Symposium on Organic Reaction Mechanism
  • Int'l Joint Research / Invited
• [Presentation] Simple Screening for High-Colibactin Producers from Clinical Samples Using Activity- Based Fluorescent Probe (2018)
  • Author(s): Kenji Watanabe
  • Organizer: U.S.-Japan Cooperative Medical Sciences Program (USJCMSP)
  • Int'l Joint Research / Invited
• [Presentation] ENZYMATIC ONE-STEP RING CONTRACTION FOR QUINOLONE BIOSYNTHESIS (2018)
  • Author(s): Kenji Watanabe
  • Organizer: 2nd China-Japan Joint Symposium on Natural Product Biosynthesis
  • Int'l Joint Research / Invited
• [Presentation] Simple Screening for High-Colibactin Producers from Clinical Samples Using Activity- Based Fluorescent Probe (2018)
  • Author(s): Kenji Watanabe
  • Organizer: 30th International Symposium on the Chemistry of Natural Products
  • Int'l Joint Research / Invited
• [Presentation] Simple Screening for High-Colibactin Producers from Clinical Samples Using Activity- Based Fluorescent Probe (2018)
  • Author(s): Kenji Watanabe
  • Organizer: 6th Biennial International Conference on New Developments in Drug Discovery from Natural Products and Traditional Medicines
  • Int'l Joint Research / Invited
• [Presentation] Crystallographic characterization of a [4+2]-cyclase binding site and mutational alteration of stereoselectivity (2018)
  • Author(s): Kenji Watanabe
  • Organizer: The 2nd International Symposium on Organic Reaction Mechanism
  • Int'l Joint Research / Invited
• [Presentation] Genetic indoctrination of Escherichia coli for finding genotoxins “colibactin” facilitating inflammation-induced colorectal cancer (2017)
  • Author(s): Kenji Watanabe
  • Organizer: US-Japan Cooperative Medical Sciences Program, 19th International Conference on Emerging Infectious Diseases in the Pacific Rim
  • Place of Presentation: Seoul, South Korea
  • Year and Date: 2017-02-07
  • Int'l Joint Research / Invited
• [Presentation] Disfavored exo-adduct formation by a modified octalin-forming Diels-Alderase (2017)
  • Author(s): 渡辺賢二
  • Organizer: China/Japan Symposium on Natural Product Biosynthesis
  • Int'l Joint Research / Invited
• [Presentation] Disfavored exo-adduct generation by a modified octalin-forming Diels-Alderase (2017)
  • Author(s): 渡辺賢二
  • Organizer: Society for Industrial Microbiology
  • Int'l Joint Research / Invited
• [Presentation] Pseurotin biosynthesis in fungus (2017)
  • Author(s): 渡辺賢二
  • Organizer: 9th US-Japan Seminar: The international Conference of Natural Product Biosynthesis
  • Int'l Joint Research / Invited
• [Presentation] Novel Genotoxins “Colibactin” Produced by E.coli and colon carcinogenesis (In Vitro, In Vivo Experiments) (2017)
  • Author(s): 渡辺賢二
  • Organizer: The 12th international conference and 5th Asian congress on environmental mutagens with the 33rd annual meeting of KSOT/KEMS
  • Int'l Joint Research / Invited
• [Presentation] Genetic indoctrination of Escherichia coli for finding genotoxins colibactin facilitating inflammation-induced colorectal cancer (2017)
  • Author(s): 渡辺賢二
  • Organizer: US-Japan Cooperative Medical Sciences Program Cancer Panel Meeting
  • Int'l Joint Research / Invited
• [Presentation] 実用的物質生産系構築にむけたゲノム情報に基づく 新規生合成システムのリデザイ (2017)
  • Author(s): 渡辺賢二
  • 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化学（新学術領域）第一回公開シンポジウム
  • Place of Presentation: 東京大学, 東京
  • Int'l Joint Research / Invited
• [Presentation] Oxidative trans-to-cis isomerization of olefin in polyketide biosynthesis (2016)
  • Author(s): Kenji Watanabe
  • Organizer: The 13th International Symposium on the Genetics of Industrial Microorganisms
  • Place of Presentation: Wuhan, People’s Republic of China
  • Year and Date: 2016-10-16
  • Int'l Joint Research / Invited
• [Book] 遺伝子・細胞から見た 応用微生物学 (2020)
  • Author(s): 阪井 康能、竹川 薫、橋本 渉、片山高嶺
  • Total Pages: 200
  • Publisher: 朝倉書店
  • ISBN: 9784254431247
• [Book] 天然物の化学Ⅱ（科学のとびら64) (2018)
  • Author(s): 上村大輔, 渡辺賢二
  • Total Pages: 192
  • Publisher: 東京化学同人
  • ISBN: 9784807915057
• [Book] 天然物生合成におけるS-アデノシルメチオニン依存性酵素により触媒されるペリ環状反応 (2018)
  • Author(s): 大橋雅生, 渡辺賢二
  • Total Pages: 6
  • Publisher: 現代化学
• [Book] Polyketide synthase and nonribosomal peptide synthetase hybrid enzymes of fungi. The Mycota XV: Physiology and Genetics (2018)
  • Author(s): Kishimoto, S., Hirayama, Y., Watanabe, K.
  • Total Pages: 7
  • Publisher: Springer
• [Book] 現代化学 (2017)
  • Author(s): 大橋雅生, 渡辺賢二
  • Total Pages: 6
  • Publisher: 東京化学同人
• [Book] BIO Clinica (2017)
  • Author(s): 渡辺賢二
  • Total Pages: 4
  • Publisher: 北隆館／ニュー・サイエンス社
• [Remarks] https://sweb.u-shizuoka-ken.ac.jp/~kenji55-lab/
• [Remarks] 渡辺賢二 研究室
  • URL: http://sweb.u-shizuoka-ken.ac.jp/~kenji55-lab/
• [Remarks] 研究内容
  • URL: http://sweb.u-shizuoka-ken.ac.jp/~kenji55-lab/
• [Remarks] WATANABE RESEARCH GROUP
  • URL: http://sweb.u-shizuoka-ken.ac.jp/~kenji55-lab/
• [Patent(Industrial Property Rights)] コリバクチン産生菌検出用プライマーセット及び検出用キット (2019)
  • Inventor(s): 渡辺賢二
  • Industrial Property Rights Holder: 渡辺賢二
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-027695
  • Filing Date: 2019
• [Patent(Industrial Property Rights)] コリバクチンおよびコリバクチン産生菌の検出方法および検出プローブ (2018)
  • Inventor(s): 渡辺賢二
  • Industrial Property Rights Holder: 渡辺賢二
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018
  • Overseas