植物由来新規ポリケタイド閉環酵素の探索と物質生産

2018 Fiscal Year Annual Research Report

• Project Area: Creation of Complex Functional Molecules by Rational Redesign of Biosynthetic Machineries
• Project/Area Number: 17H05435
• Research Institution: University of Toyama
• Principal Investigator: 森田 洋行 富山大学, 和漢医薬学総合研究所, 教授 (20416663)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Keywords: 生合成 / 酵素工学 / X線結晶構造解析

Outline of Annual Research Achievements

（1）キダチアロエ由来のⅢ型ポリケタイド合成酵素（PKS）であるオクタケタイド合成酵素（OKS）とアサ由来オリベトール酸閉環酵素（OAC）の共反応液に、ブチリルCoAとマロニルCoAを基質として作用させた場合の反応生成物について評価した。その結果、まず、OKSが1分子のブチリルCoAと7分子のマロニルCoAから新規パイロン誘導体を生産することが判明した。さらに、興味深いことに、OACがOKSの酵素反応液に共存すると、その新規化合物の生成量が約1.5倍増加することが確認された。このことから、OACは、プロピル基を有するポリケタイドCoAを基質として受け入れることが明らかとなった。これまでOACは、ペンチル基を有するポリケタイドCoA以外は基質として受け入れることができないと考えられていたが、本結果は、OACが他の長さのアシル基を有するポリケタイドCoAを基質として受け入れる可能性を示している。現在、他の長さのアシル基を有するポリケタイドCoAに対するOACの基質特異性について再評価を行っているところである。
（2）昨年度に引き続き、OACのホモログ酵素と目される植物由来AtHS1及びPOP-3のポリケタイド閉環酵素としての機能について検討したが、いずれの基質を用いても、酵素反応生成物を確認することができなかった。現在、これについては、酵素反応条件の最適化を行っているところである。
（3）領域横断型共同研究として、当該研究領域の研究者らが新たに見い出した酵素の結晶構造の取得と詳細な触媒機構の解明を試みた。その結果、アセチルCoAとマロニルCoAを基質としてオルセリン酸の生合成に関与すると想定されるエゾムラサキツツジ由来新規Ⅲ型PKSのX線結晶構造の取得に成功し，その基質特異性や生産物特異性が，活性中心キャビティーの形状と大きさに極めて依存していることを明らかにした。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Lignans with melanogenesis effects from Premna serratifolia wood (2019)
  • Author(s): S. Woo, S. Hoshino, C.P. Wong, N.N. Win, M.D. Awoufack, Prema, H. Ngwe, H. Zhang, F. Hayashi, I. Abe, H. Morita
  • Journal Title: Fitoterapia Volume: 133 Pages: 35-42
  • DOI: 10.1016/j.fitote.2018.12.008
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A new tetrahydrofuran type lignan from Premna serratifolia wood (2019)
  • Author(s): S. Woo, C.P. Wong, N.N. Win, S. Hoshino, Prema, H. Ngwe, I. Abe, H. Morita
  • Journal Title: Nat Prod Commun Volume: 14 Pages: 113-116
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Bis-iridoid and iridoid glycosides: Viral protein R inhibitors from Picrorhiza kurroa collected in Myanmar (2019)
  • Author(s): N.N. Win, T. Kodama, K.Z.W. Lae, Y.Y. Win, H. Ngwe, I Abe, H. Morita
  • Journal Title: Fitoterapia Volume: 134 Pages: 101-107
  • DOI: 10.1016/j.fitote.2019.02.016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Brominated diphenyl ethers including a new tribromoiododiphenyl ether from a Vietnamese marine sponge Arenosclera sp. and their antibacterial activities (2019)
  • Author(s): D. Ki, M.D. Awouafack, C.P. Wong, H.M. Nguyen, Q. M. Thai, L.H.T. Nu, H. Morita
  • Journal Title: Chem Biodivers Volume: 16 Pages: e1800593
  • DOI: 10.1002/cbdv.201800593
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Two new pyrrolo-2-aminoimidazoles from a Myanmarese marine sponge, Clathria prolifera (2018)
  • Author(s): S. Woo, N.N. Win, C.P. Wong, T. Ito, S. Hoshino, H. Ngwe, A.A. Aye, N.M. Han, H. Zhang, F. Hayashi, I. Abe, H. Morita
  • Journal Title: J Nat Med Volume: 72 Pages: 803-807
  • DOI: 10.1007/s11418-018-1205-y
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Mirilactams C-E, novel polycyclic macrolactams isolated from combined-culture of Actinosynnema mirum NBRC 14064 and mycolic acid-containing bacterium (2018)
  • Author(s): S. Hoshino, M. Ozeki, C.P. Wong, H. Zhang, F. Hayashi, T. Awakawa, H. Morita, H. Onaka, I. Abe
  • Journal Title: Chem Pharm Bull Volume: 66 Pages: 660-667
  • DOI: 10.1248/cpb.c18-00143
  • Peer Reviewed
• [Journal Article] A new coumaronochromone and a new alkanoyl-dihydrofuranoflavone glycoside from Eriosema robustum (Fabaceae) (2018)
  • Author(s): M.D. Awouafack, C.P. Wong, P. Tane, H. Morita
  • Journal Title: Phytochem Lett Volume: 27 Pages: 20-24
  • DOI: 10.1016/j.phytol.2018.06.024
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Huperphlegmines A and B, two novel Lycopodium alkaloids with an unprecedented skeleton from Huperzia phlegmaria, and their acetylcholinesterase inhibitory activities (2018)
  • Author(s): H.T. Nguyen, H.T. Doan, D.V. Ho, K. T. Pham, A. Raal, H. Morita
  • Journal Title: Fitoterapia Volume: 129 Pages: 267-271
  • DOI: 10.1016/j.fitote.2018.07.016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Pogostemins A-C, three new cytotoxic meroterpenoids from Pogostemon auricularius (2018)
  • Author(s): H.T. Nguyen, L.T.T. Tran, D.V. Ho, D.V. Le, A. Raal, H. Morita
  • Journal Title: Fitoterapia Volume: 130 Pages: 100-104
  • DOI: 10.1016/j.fitote.2018.08.015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Catenulobactins A and B, heterocyclic peptides from culturing Catenuloplanes sp. with mycolic acid-containing bacterium (2018)
  • Author(s): S. Hoshino, M. Ozeki, T. Awakawa, H. Morita, H. Onaka, I. Abe
  • Journal Title: J Nat Prod Volume: 81 Pages: 2106-2110
  • DOI: 10.1021/acs.jnatprod.8b00261
  • Peer Reviewed
• [Journal Article] A new Limonoid from Chisocheton paniculatus Fruit collected in Vietnam and its NO production inhibitory activity (2018)
  • Author(s): H.T. Nguyen, H.V. Duc, A. Raal, H. Morita
  • Journal Title: Nat Prod Commn Volume: 13 Pages: 1255-1257
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Characterization of two novel plant type III polyketide synthase from Evodia rutaecarpa. (2019)
  • Author(s): Morita H.
  • Organizer: 2nd China-Japan Joint Symposium on Natural Product Biosynthesis
  • Int'l Joint Research
• [Presentation] X-ray crystal structure analysis of agarwood type III PKS PECPS (2019)
  • Author(s): Wong C. P., Liu Q., Shi S., Morita H.
  • Organizer: 日本薬学会第139年会
• [Presentation] 植物由来新規ポリケタイド閉環酵素の探索と物質生産 (2018)
  • Author(s): 森田洋行
  • 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「生合成リデザイン」 第４回公開シンポジウム
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): Morita H.
  • 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): Morita H.
  • Organizer: Seminar Nasional Kefarmasian Indonesian Pharmaceutical Update
  • Int'l Joint Research / Invited
• [Presentation] Discovery of new lignans from wood of Premna integrifolia collected in Myanmar (2018)
  • Author(s): Woo S., Win N. N., Morita H.
  • Organizer: 第５回富山・バーゼル医薬品研究開発シンポジウム
• [Presentation] Combinatorial biosynthesis of aloesone analog by exploiting a type III polyketide synthase and olivetolic cyclase (2018)
  • Author(s): Woo B., Morita H.
  • Organizer: 第５回富山・バーゼル医薬品研究開発シンポジウム
• [Presentation] Potent HIV-1 Vpr inhibitors from Kaempferia pulchra Rhizomes Collected in Myanmar (2018)
  • Author(s): Win N. N., Morita H.
  • Organizer: 第５回富山・バーゼル医薬品研究開発シンポジウム
• [Presentation] New sesquiterpene quinones from a Vietnamese marine sponge Spongia sp.and their biological activities (2018)
  • Author(s): Nguyen H.M., Morita H.
  • Organizer: 第５回富山・バーゼル医薬品研究開発シンポジウム
• [Presentation] Combinatorial biosynthesis of an unnatural novel aloeson analog by exploiting type III polyketide synthase and olivetolic acid cyclase (2018)
  • Author(s): Morita H.
  • Organizer: The Japan-German Biosynthesis Symposium
  • Int'l Joint Research
• [Presentation] Crystal structure analysis of a new type III polyketide synthase that catalyzes one pot formation of the C6-C-5-C6 diarlypentanoid (2018)
  • Author(s): Liu Q., Wong C. P., Zhao Y., Shi S., Morita H.
  • Organizer: The 3rd International Symposium on Toyama-Asia-Africa Pharmaceutical Network
• [Presentation] Two new quassinoids isolated from a medicinal plant Picrasma javanica of Myanmar (2018)
  • Author(s): Prema, Wong C. P., Awouafack M. D., Win N. N., Ngwe H., Morita H.
  • Organizer: The 3rd International Symposium on Toyama-Asia-Africa Pharmaceutical Network
• [Presentation] New sesquiterpene aminoquinones from a Vietnamese Spongia sp. and their biological activities (2018)
  • Author(s): Nguyen H. M., Ito T., Morita H.
  • Organizer: 第65回日本生薬学会年会
• [Presentation] Melanogenesis is inhibitor tetrahydrofuran lignans from Premna integrifolia collected in Myanmar (2018)
  • Author(s): Win N. N., Woo S., Prema, Wong C. P., Ito T., Ngwe H., Morita H.
  • Organizer: 第65回日本生薬学会年会
• [Presentation] Two distinct substrate binding modes for the normal and reverse prenylation by AmbP3 (2018)
  • Author(s): Wong C. P., Awakawa T, Nakashima Y., Mori T., Liu X., Morita H., Abe I.
  • Organizer: 第65回日本生薬学会年会
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): Morita H.
  • Organizer: Seminar on Ho Chi Minh Medicine and Pharmacy University
  • Invited
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): Morita H.
  • Organizer: Seminar on Cantho University
  • Invited
• [Presentation] Synthesis of unnatural compounds by exploiting secondary metabolite enzyme (2018)
  • Author(s): Morita H.
  • Organizer: The 60th Anniversary of School of Chinese Materia Medica Conference
  • Invited
• [Presentation] Synthesis of unnatural compounds by enzyme engineering (2018)
  • Author(s): Morita H.
  • 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): Woo B., Hoshino S., Win N. N., Abe I., Morita H.
  • Organizer: The 10th KSP-JSP-CSP Joint Symposium on Pharmacognosy
  • Int'l Joint Research
• [Presentation] Anti-melanin deposition activity of a new tetrahydrofuran lignin from Premna serratifolia wood (2018)
  • Author(s): Woo S., Wong C. P., Win N. N., Hoshino S., Prema, Ngwe H., Abe I., Morita H.
  • Organizer: The 10th KSP-JSP-CSP Joint Symposium on Pharmacognosy
  • Int'l Joint Research
• [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