Modification of polyketide synthase and elucidation of enzyme reaction which is difficult in chemical synthesis, and creation of novel natural products

• Project/Area Number: 16H04905
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Takahashi Shunji 国立研究開発法人理化学研究所, 環境資源科学研究センター, ユニットリーダー (30311608)
• Co-Investigator(Kenkyū-buntansha): 奥村 英夫 公益財団法人高輝度光科学研究センター, タンパク質結晶解析推進室, 研究員 (90377903)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2018: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2017: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2016: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
• Keywords: リベロマイシン / ポリケチド合成酵素 / 異種発現 / 放線菌 / 機能ドメイン改変 / ポリケチド / ドメイン改変 / 二次代謝産物 / 生合成 / 微生物 / BACベクター

Outline of Final Research Achievements

Natural products are important sources for drug discovery because of their diverse structures and strong biological activities. Understanding and utilization of unique enzymes involved in biosynthetic reactions of natural compounds is an important issue. In this study, we focused on enzymes involved in reveromycin A (RM-A) biosynthesis. Esterification of the tertiary hydroxyl group at the final stage of RM-A biosynthesis is a very difficult reaction in organic synthesis. With this research grant, we succeeded in crystal structure analysis of three enzymes responsible for the reaction.

Academic Significance and Societal Importance of the Research Achievements

ポリケチド化合物群は貴重な創薬探索源であり、ポリケチド合成酵素が有する機能ドメインとモジュール構造を改変することは、重要な学術課題である。また、生合成遺伝子クラスター全長を取得し、遺伝子改変を行う意義は極めて大きい。さらに、これらの化合物の構造修飾機構を明らかにすることも重要である。特にRM-A生合成過程には、有機合成的には極めて困難な3級水酸基のエステル化が存在する。これら生合成酵素群の反応機構解明は、難溶性化合物の水溶性向上及びプロドラッグ化手法の開発につながる。新しい学術知見をもとに、天然化合物の創出基盤を構築することによって、社会に貢献する創薬シード導出が期待される。

Research Products

• [Int'l Joint Research] University of Kentucky(米国)
• [Journal Article] β-carboline biomediators induce reveromycin production in Streptomyces sp. SN-593. (2019)
  • Author(s): Panthee S., Takahashi S.*, Hayashi T., Shimizu T, and Osada H.
  • Journal Title: Sci. Rep. Volume: 9 Issue: 1 Pages: 5802-5802
  • DOI: 10.1038/s41598-019-42268-w
  • Peer Reviewed / Open Access
• [Journal Article] Pentaminomycins A and B, hydroxyarginine-containing cyclic pentapeptides from Streptomyces sp. RK88-1441. (2018)
  • Author(s): Jang J.-P., Hwang G.J., Kwon M.C., Ryoo I.-J., Jang M., Takahashi S., Ko S.K., Osada H., Jang J.H., and Ahn J.S.
  • Journal Title: J. Nat. Prod. Volume: 81 Issue: 4 Pages: 806-810
  • DOI: 10.1021/acs.jnatprod.7b00882
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Aturanosides A and B, glycosylated anthraquinones with antiangiogenic activity from a soil-derived Streptomyces species. (2018)
  • Author(s): Jang J.-P., Hwang G.J., Jang M., Takahashi S., Ko S.K., Osada H., Jang J.H., and Ahn J.S.
  • Journal Title: J. Nat. Prod. Volume: 81 Issue: 9 Pages: 2004-2009
  • DOI: 10.1021/acs.jnatprod.8b00307
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Control of the stereochemical course of [4+2] cycloaddition during trans-decalin formation by Fsa2-family enzymes (2018)
  • Author(s): Kato N, Nogawa T, Takita R, Kinugasa K, Kanai M, Uchiyama M, Osada H, Takahashi S
  • Journal Title: Angew Chem Int Ed Engl Volume: 57 Issue: 31 Pages: 9754-9758
  • DOI: 10.1002/anie.201805050
  • Peer Reviewed
• [Journal Article] Catenulisporolides, glycosylated triene macrolides from the chemically underexploited actinomycete Catenulispora species. (2018)
  • Author(s): Son S., Hong Y.S., Futamura Y., Jang M., Lee J.K., Heo K.T., Ko S.K., Lee J.S., Takahashi S., Osada H., Jang J.H., Ahn J.S.
  • Journal Title: Org. Lett. Volume: 20 Issue: 22 Pages: 7234-7238
  • DOI: 10.1021/acs.orglett.8b03160
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Biosynthesis of Quinolidomicin, the Largest Known Macrolide of Terrestrial Origin: Identification and Heterologous Expression of a Biosynthetic Gene Cluster over 200 kb (2018)
  • Author(s): Hashimoto Takuya、Hashimoto Junko、Kozone Ikuko、Amagai Keita、Kawahara Teppei、Takahashi Shunji、Ikeda Haruo、Shin-ya Kazuo
  • Journal Title: Organic Letters Volume: 20 Issue: 24 Pages: 7996-7999
  • DOI: 10.1021/acs.orglett.8b03570
  • Peer Reviewed / Open Access
• [Journal Article] Wakodecalines A and B, new decaline metabolites isolated from a fungus Pyrenochaetopsis sp. RK10-F058 (2018)
  • Author(s): Nogawa T, Kato N, Shimizu T, Okano A, Futamura Y, Takahahi S, Osada H
  • Journal Title: The Journal of Antibiotica Volume: 71 Issue: 1 Pages: 123-128
  • DOI: 10.1038/ja.2017.103
  • Peer Reviewed
• [Journal Article] Kinanthraquinone, a new anthraquinone carboxamide isolated from Streptomyces reveromyceticus SN-593-44 (2018)
  • Author(s): Takagi H, Nogawa T, Futamura Y, Takahashi S, Osada H
  • Journal Title: The Journal of Antibiotics Volume: 71 Issue: 4 Pages: 480-482
  • DOI: 10.1038/s41429-017-0020-0
  • Peer Reviewed
• [Journal Article] Biosynthesis and structure activity relationship studies of okaramines that target insect glutamate-gated chloride channels (2018)
  • Author(s): Kato Naoki、Furutani Shogo、Otaka Junnosuke、Noguchi Akira、Kinugasa Kiyomi、Kai Kenji、Hayashi Hideo、Ihara Makoto、Takahashi Shunji、Matsuda Kazuhiko、Osada Hiroyuki
  • Journal Title: ACS Chemical Biology Volume: 13 Issue: 3 Pages: 561-566
  • DOI: 10.1021/acschembio.7b00878
  • Peer Reviewed
• [Journal Article] Identification of a gene cluster for telomestatin biosynthesis and heterologous expression using a specific promoter in a clean host. (2017)
  • Author(s): 4.Amagai, K., Ikeda, H., Hashimoto, J., Kozone, I., Izumikawa, M., Kudo, F., Eguchi, T., Nakamura, T., Osada, H., Takahashi, S., Shin-ya
  • Journal Title: Sci. Rep. Volume: 7 Issue: 1 Pages: 3382-3382
  • DOI: 10.1038/s41598-017-03308-5
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of giant modular PKSs provides insight into genetic mechanism for structural diversification of aminopolyol polyketides (2017)
  • Author(s): Zhang, L., Hashimoto, T., Qin, B., Hashimoto, J., Kozone, I., Kawahara, T., Okada, M., Awakawa, T., Ito, T., Asakawa, Y., Ueki, M., Takahashi, S., Osada, H., Wakimoto, T., Ikeda, H., Shin-ya, K., Abe, I.
  • Journal Title: Angew. Chem. Int. Ed. Volume: 56 Issue: 7 Pages: 1740-1745
  • DOI: 10.1002/anie.201611371
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Octaminomycins A and B, Cyclic Octadepsipeptides Active against Plasmodium falciparum. (2017)
  • Author(s): Jang JP, Nogawa T, Futamura Y, Shimizu T, Hashizume D, Takahashi S, Jang JH, Ahn JS, Osada H
  • Journal Title: J Nat Prod Volume: 80 Issue: 1 Pages: 134-140
  • DOI: 10.1021/acs.jnatprod.6b00758
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Development of a terpenoid-production platform in Streptomyces reveromyceticus SN-593 (2017)
  • Author(s): Khalid A、Takagi H、Panthee S、Muroi M、Chappell J、Osada H、Takahashi S
  • Journal Title: ACS Synthetic Biology Volume: 6 Issue: 12 Pages: 2339-2349
  • DOI: 10.1021/acssynbio.7b00249
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Identification of a novel sesquiterpene biosynthetic machinery involved in astellolide biosynthesis. (2016)
  • Author(s): Shinohara Y, Takahashi S, Osada H, Koyama Y
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 32865-32865
  • DOI: 10.1038/srep32865
  • Peer Reviewed / Open Access
• [Journal Article] A crotonyl-CoA reductase-carboxylase independent pathway for assembly of unusual alkylmalonyl-CoA polyketide synthase extender units. (2016)
  • Author(s): Ray L., Valentic T., Miyazawa T., Withall D.M., Song L., Milligan J.C., Osada H., Takahashi S., Tsai S.-C., and Challis G.L.
  • Journal Title: Nat. Commun Volume: 7 Issue: 1 Pages: 13609-13609
  • DOI: 10.1038/ncomms13609
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] リベロマイシン生合成に関わるRevKの結晶構造解析 (2019)
  • Author(s): 奥村英夫、鬼頭奈央子、佐藤裕美、熊坂崇、長田裕之、高橋俊二
  • Organizer: 農芸化学会
• [Presentation] リベロマイシン生合成に関わるサクシニル化酵素群の相互作用解析 (2019)
  • Author(s): 鬼頭奈央子、佐藤裕美、奥村英夫、熊坂崇、長田裕之、高橋俊二
  • Organizer: 農芸化学会
• [Presentation] Development of a terpenoid-production platform in Streptomyces reveromyceticus SN-593 (2019)
  • Author(s): Shunji Takahashi
  • Organizer: 2nd China-Japan Joint Symposium on Natural Product Biosynthesis
  • Int'l Joint Research
• [Presentation] 鬼頭奈央子、佐藤裕美、奥村英夫、熊坂崇、長田裕之、高橋俊二 (2018)
  • Author(s): リベロマイシン生合成に関わるサクシニル化酵素群の機能解析
  • Organizer: 放線菌学会
• [Presentation] Kinanthraquinone生合成遺伝子クラスターの解析 (2018)
  • Author(s): 高尾 理沙、鬼頭 奈央子、高木 海、野川 俊彦、長田 裕之、高橋 俊二
  • Organizer: 放線菌学会
• [Presentation] 立体選択的デカリン形成を担うFsa2ファミリーDiels-Alderaseの機能解析 (2018)
  • Author(s): 加藤直樹、野川俊彦、衣笠清美、高橋俊二、長田裕之
  • Organizer: 農芸化学会
• [Presentation] リベロマイシン生合成に関わるサクシニル化酵素群の機能解析 (2018)
  • Author(s): 鬼頭奈央子、高橋俊二、佐藤裕美、奥村英夫、熊坂崇、長田裕之
  • Organizer: 農芸化学会
• [Presentation] 糸状菌Pyrenochaetopsis sp. RK10-F058より単離した新規デカリン骨格含有二次代謝産物wakodecaline Cの構造 (2018)
  • Author(s): 野川俊彦、加藤直樹、清水猛、岡野亜紀子、二村友史、高橋俊二、長田裕之
  • Organizer: 農芸化学会
• [Presentation] 新規アントラキノンkinanthraquinoneの同定と生物活性評価 (2018)
  • Author(s): 髙木 海、野川俊彦、二村友史、長田裕之、高橋俊二
  • Organizer: 農芸化学会
• [Presentation] 麹菌Aspergillus oryzaeが有する新奇セスキテルペン生合成経路の解明 (2017)
  • Author(s): 篠原靖智，高橋俊二，長田裕之，小山泰二
  • Organizer: 農芸化学会
  • Place of Presentation: 京都（京都女子大）
  • Year and Date: 2017-03-17
• [Presentation] リベロマイシン生合成に関わる酸化還元酵素RevMの機能解析 (2017)
  • Author(s): 鬼頭奈央子、高橋俊二、宮澤岳、前島希、田中美帆、室井誠、奥村英夫、熊坂崇、長田裕之
  • Organizer: 農芸化学会
  • Place of Presentation: 京都（京都女子大）
  • Year and Date: 2017-03-17
• [Presentation] Development of terpenoid production platform using Streptomyces reveromyceticus (2017)
  • Author(s): Ammara Khalid, Hiroshi Takagi, Suresh Panthee, Hiroyuki Osada, and Shunji Takahashi.
  • Organizer: 農芸化学会
  • Place of Presentation: 京都（京都女子大）
  • Year and Date: 2017-03-17
• [Presentation] リベロマイシン生合成に関わるRevMの機能解析 (2017)
  • Author(s): 鬼頭奈央子、高橋俊二、佐藤裕美、奥村英夫、熊坂崇、長田裕之
  • Organizer: 放線菌学会
• [Presentation] Regulation of Reveromycin Biosynthetic Gene Cluster by Small Molecule (2017)
  • Author(s): Shunji Takahashi
  • Organizer: 18th International Symposium on the Biology of Actinomycetes
  • Int'l Joint Research
• [Presentation] アミノ酸置換による新規テロメスタチン類縁体の創製 (2016)
  • Author(s): 天貝 啓太, 池田 治生, 新家 一男, 高橋 俊二
  • Organizer: 放線菌学会
  • Place of Presentation: 東京大学
  • Year and Date: 2016-09-08
• [Remarks] 天然物生合成研究ユニット
  • URL: http://www.csrs.riken.jp/jp/labs/npbu/index.html
• [Remarks] 天然物生合成研究ユニット
  • URL: http://www.npbrt.riken.jp/