Study for glycyrrhizin production in Glycyrrhiza plant.

• Project/Area Number: 16K08301
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Natural medicines
• Research Institution: Health Sciences University of Hokkaido
• Principal Investigator: Kojomo Mareshige 北海道医療大学, 薬学部, 准教授 (80342781)
• Co-Investigator(Kenkyū-buntansha): 関 光 大阪大学, 工学研究科, 准教授 (30392004) ; 金 尚永 北海道医療大学, 薬学部, 講師 (70624287)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: カンゾウ / Glycyrrhiza / グリチルリチン酸 / トリテルペノイド / 生合成 / 植物 / 薬学

Outline of Final Research Achievements

Glycyrrhiza uralensis (Licorice) is used as a natural herbal drug. Glycyrrhizin, a triterpenoid saponin derived from the underground parts, is a major bioactive compound that has several pharmacological activities. Licorice also produces other triterpenoids, including soyasaponins. Recent studies have revealed various oxidosqualene cyclases and cytochrome P450 monooxygenases required for the biosynthesis of triterpenoids in licorice.
[1] CYP88D6 has a key role in the glycyrrhizin biosynthetic pathway. We found that over-expression of the CYP88D6 gene can cause a marked accumulation of glycyrrhizin and 11-oxo--amyrin in the transformed hairy roots. [2] CYP716A179 expressed predominantly in tissue-cultured stolons of licorice, functions as a triterpene C-28 oxidase in the biosynthesis of oleanolic acid and betulinic acid. [3] The basic helix-loop-helix transcription factor GubHLH3 positively regulates soyasaponin biosynthetic genes in licorice.

Academic Significance and Societal Importance of the Research Achievements

医薬品原料として高品質なカンゾウを社会に供するためには、グリチルリチン酸生成に関するトリテルペノイド生合成機構を解明することが必要である。本研究から、トリテルペノイド化合物の生合成酵素遺伝子の解析、発現制御を明らかにした。このような基礎データのさらなる蓄積により、薬用成分含有量の高いカンゾウを品種改良して社会に供給することが可能となる。また培養細胞による医薬品、機能性成分の生産も可能となる。

Research Products

• [Journal Article] The Basic Helix-Loop-Helix Transcription Factor GubHLH3 Positively Regulates Soyasaponin Biosynthetic Genes in Glycyrrhiza uralensis (2018)
  • Author(s): Tamura Keita、Yoshida Koki、Hiraoka Yasuko、Sakaguchi Daiki、Chikugo Ayaka、Mochida Keiichi、Kojoma Mareshige、Mitsuda Nobutaka、Saito Kazuki、Muranaka Toshiya、Seki Hikaru
  • Journal Title: Plant and Cell Physiology Volume: 59 Issue: 4 Pages: 778-791
  • DOI: 10.1093/pcp/pcy046
  • Peer Reviewed
• [Journal Article] CYP716A179 functions as a triterpene C-28 oxidase in tissue-cultured stolons of Glycyrrhiza uralensis. (2017)
  • Author(s): Tamura, K., Seki, H., Suzuki, H., Kojoma, M., Saito, K. and Muranaka, T.: CYP716A179 functions as a triterpene C-28 oxidase in tissue-cultured stolons of Glycyrrhiza uralensis.
  • Journal Title: Plant Cell Rep. Volume: 36 Issue: 3 Pages: 437-445
  • DOI: 10.1007/s00299-016-2092-x
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Glycyrrhizin production in hairy root cultures of Glycyrrhiza uralensis induced triterpenoid biosynthetic gene (2016)
  • Author(s): M Kojoma, H Seki, K Ohyama, SY Kim, T Muranaka
  • Journal Title: Planta Medica Volume: 82 Issue: S 01 Pages: S1-S381
  • DOI: 10.1055/s-0036-1596798
  • Peer Reviewed / Open Access
• [Presentation] カンゾウと微生物の共生に関する研究 (2017)
  • Author(s): 中尾隆寛、千々岩諒太、石丸幹二、有馬進、高上馬希重
  • Organizer: 第8回甘草に関するシンポジウム
• [Presentation] Identification of beta-amyrin 28-oxidase in Glycyrrhiza uralensis (2016)
  • Author(s): Keita Tamura, Hikaru Seki, Hideyuki Suzuki, Mareshige Kojoma, Toshiya Muranaka
  • Organizer: 日本生物工学会年次大会
  • Place of Presentation: 富山県・富山市
  • Year and Date: 2016-09-28
• [Presentation] 薬用植物カンゾウのトリテルペノイド生合成制御に関わる転写因子の探索 (2016)
  • Author(s): 田村啓太、關光、吉田光輝、平岡靖子、持田恵一、鈴木秀幸、高上馬希重、光田展隆、高木優、斉藤和季、村中俊哉
  • Organizer: 第34回日本植物細胞分子生物学会（上田）大会
  • Place of Presentation: 長野県・上田市
  • Year and Date: 2016-09-01
• [Presentation] Identification of beta-amyrin 28-oxidase in Glycyrrhiza uralensis (2016)
  • Author(s): Keita Tamura, Hikaru Seki, Hideyuki Suzuki, Mareshige Kojoma, Toshiya Muranaka
  • Organizer: 55th Annual Meeting of the Phytochemical Society of North America
  • Place of Presentation: University of California, Davis, USA
  • Year and Date: 2016-08-06
  • Int'l Joint Research
• [Presentation] Glycyrrhizin production in hairy root cultures of Glycyrrhiza uralensis induced triterpenoid biosynthetic gene. (2016)
  • Author(s): Mareshige Kojoma, Hikaru Seki, Kiyoshi Ohyama, Sang-Yong Kim, Toshiya Muranaka
  • Organizer: 9th Joint Natural Products Conference 2016
  • Place of Presentation: Copenhagen, Denmark
  • Year and Date: 2016-07-24
  • Int'l Joint Research