Engineering of DABB-family enzymes to novel polyketide cyclase

• Project/Area Number: 17H02203
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: University of Toyama
• Principal Investigator: Morita Hiroyuki 富山大学, 学術研究部薬学・和漢系, 教授 (20416663)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2019: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2018: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2017: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
• Keywords: 生合成 / 酵素工学 / ポリケタイド / アルカロイド

Outline of Final Research Achievements

In order to generate new compounds, site-directed mutagenesis studies were carried out on olivetolic acid cyclase (OAC) from Cannavis sativa, boiling stable protein from Populus tremula, and At5g22580 from Arabidopsis thaliana. As a result, a substitution of valine 59 with isoleucine in OAC led to production of new olivetolic analogs by accepting polyketide-CoAs with C10 to C12 fatty acyl moieties as substrates, respectively.

Academic Significance and Societal Importance of the Research Achievements

天然化合物の生合成に関わる生合成酵素を，本来の生物とは異なった生物内において発現することで，目的化合物を作ることが可能になっている。本手法を用いれば微量な天然化合物や新規化合物を効率的に生産することも可能である。今回我々は，医薬品としても重要なテトラヒドロカンナビノイドの生合成において鍵中間体の生産に関わる酵素に変異を導入することで，この酵素に新たな化合物を生産する能力を付与することに成功した。本機能改変酵素をテトラヒドロカンナビノイドの生合成に人為的に組み込むことで，テトラヒドロカンナビノイド類似化合物の安価で容易な創出が実現することが期待される。

Research Products

• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2019)
  • Author(s): H. Morita
  • Organizer: 2019 International Conference of The Plant Resources Society of Korea
  • Int'l Joint Research / Invited
• [Presentation] Engeering of olivetolic cyclase to generate new compounds (2019)
  • Author(s): H. Morita
  • Organizer: JST-CRSC「日中分野別ハイレベル研究者交流会2019」医薬・健康編
  • Int'l Joint Research / Invited
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): H. Morita
  • Organizer: 11th International Symposium on Chromatography of Natural Products
  • Int'l Joint Research / Invited
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): H. Morita
  • Organizer: Seminar Nasional Kefarmasian Indonesian Pharmaceutical Update
  • Int'l Joint Research / Invited
• [Presentation] Combinatorial biosynthesis of aloesone analog by exploiting a type III polyketide synthase and olivetolic cyclase (2018)
  • Author(s): B. Woo., H. Morita
  • Organizer: 第５回富山・バーゼル医薬品研究開発シンポジウム
  • Int'l Joint Research
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): H. Morita
  • Organizer: Seminar on Ho Chi Minh Medicine and Pharmacy University
  • Invited
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): H. Morita
  • Organizer: Seminar on Cantho University
  • Invited
• [Presentation] Synthesis of unnatural compounds by exploiting secondary metabolite enzyme (2018)
  • Author(s): H. Morita
  • Organizer: The 60th Anniversary of School of Chinese Materia Medica Conference
  • Int'l Joint Research / Invited
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): H. Morita
  • Organizer: The 10th KSP-JSP-CSP Joint Symposium on Pharmacognosy
  • Int'l Joint Research / Invited
• [Presentation] Combinatorial biosynthesis of aloesone analog by exploiting a type III polyketide synthase and olivetolic acid cyclase (2018)
  • Author(s): B. Woo, S. Hoshino, N. N. Win, I. Abe, H. Morita
  • Organizer: The 10th KSP-JSP-CSP Joint Symposium on Pharmacognosy
  • Int'l Joint Research
• [Presentation] Characterization of two novel plant type III polyketide synthases from Evodia rutaecarpa (2017)
  • Author(s): Morita Hiroyuki
  • Organizer: 9th US-Japan Seminar on the biosynthesis of Natural Prodcuts
  • Invited
• [Presentation] Combinatorial biosynthesis of aloeson analog by exploiting a type III polykeitde synthase and olivetolic acid cyclase (2017)
  • Author(s): Ju Ye、Hoshino Shotaro、Win Nwet Nwet、Yang Xinmei、Abe Ikuro、Morita Hiroyuki
  • Organizer: The 2nd International Symposium on Toyama-Asia-Africa Pharmaceutical Network
  • Int'l Joint Research
• [Presentation] 2-Alkylquinolone alkaloid biosynthesis in the medicinal plant Evodia rutaecarpa (2017)
  • Author(s): Morita Hiroyuki
  • Organizer: International Symposium on Strategy for the future of Natural Product Resources
  • Int'l Joint Research / Invited
• [Presentation] Combinatorial biosynthesis of aloesone analog by exploiting a type III polyketide synthase and olivetolic acid cyclase (2017)
  • Author(s): Woo Bokyoung、Ju Ye、Yang Xinmei、Win Nwet Nwet、Morita Hiroyuki
  • Organizer: International Symposium on Strategy for the future of Natural Product Resources
  • Int'l Joint Research
• [Presentation] Combinatorial biosynthesis of an unnatural novel aloeson analog by exploiting type III polyketide synthase and olivetolic acid cyclase (2017)
  • Author(s): Morita Hiroyuki
  • Organizer: The 1st Japan-China Biosynthsis seminar
  • Invited
• [Presentation] Combinatorial biosynthesis of aloesone analog by exploiting a type III polyketide synthase and olivetolic acid cyclase (2017)
  • Author(s): Woo Bokyoung、Ju Ye、Yang Xinmei、Win Nwet Nwet、Morita Hiroyuki
  • Organizer: International Symposium on Scientific Research of Traditional Medicine
  • Int'l Joint Research
• [Presentation] Manipulation of plant polyketide-producing enzymes to produce new compounds (2017)
  • Author(s): Morita Hiroyuki
  • Organizer: Seminar in Jinan University
  • Invited
• [Presentation] Manipulation of plant polyketide-producing enzymes to produce new compounds (2017)
  • Author(s): Morita Hiroyuki
  • Organizer: Seminar in South China Institute of Oceanology
  • Invited
• [Remarks] 富山大学和漢医薬学総合研究所 天然物化学分野
  • URL: http://www.inm.u-toyama.ac.jp/napc/index.html