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Production of natural rubber in vitro from reconstituted-rubber synthase complex on rubber particles with Escherichia coli cell-free translation system

Research Project

Project/Area Number 18K05428
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 38030:Applied biochemistry-related
Research InstitutionSojo University (2020-2021)
Tohoku University (2018-2019)

Principal Investigator

Kojima Kouji  崇城大学, 生物生命学部, 准教授 (80457382)

Project Period (FY) 2018-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2020: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2018: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Keywords天然ゴム / 無細胞翻訳系 / パラゴムノキ / ゴム粒子 / 大腸菌 / イソプレノイド / ラテックス / 無細胞翻訳 / 酵素複合体の再構築 / イソプレン / cis-prenyltransferase / Hevea brasilliensis
Outline of Final Research Achievements

Natural rubber is a polymer of isoprene, and its biosynthesis has been thought to take place on rubber particles in the latex of Hevea brasilliensis. We have identified protein factors on the rubber particle and elucidated their roles. In the process, we have successfully reconstituted a protein complex containing cis-prenyl transferase on rubber particles using a cell-free translation system derived from wheat germ, which allowed us to produce polyisoprene in vitro with a molecular weight of 1,000,000 (Yamashita et al. al. 2016). In this study, we found that the prokaryotic cell-free translation system can be used to synthesize natural rubber synthase complexes and reconstitute them on rubber particles without relying on special factors derived from eukaryotic organisms.

Academic Significance and Societal Importance of the Research Achievements

天然ゴム生合成の機構の理解がさらに深まれば、今後高品質で高生産な天然ゴムの人工的な合成手法が確立できることが期待できる。また、合成酵素の合成や再構築には、複雑な因子の関与を必要としないことも考慮し、大腸菌などの原核生物の無細胞翻訳系を用いれば、安価な天然ゴム合成系の開発が可能になる。天然ゴム合成や複合体構築に関して、各因子の相互作用の重要性と合成活性のとの相関の解明から、今回実現が不可能であった複合体の単離や立体構造の解明に向けて一歩ずつ近づくことが可能になってきた。このように、本研究で得られる天然ゴム合成機構に関する理解は、学術(工学系、農学系)や産業へと幅広く応用されることが期待される。

Report

(5 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (17 results)

All 2022 2021 2020 2019 2018

All Journal Article (5 results) (of which Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (12 results) (of which Int'l Joint Research: 3 results,  Invited: 1 results)

  • [Journal Article] Irradiation of sound waves to Aspergillus kawachii and the characterization of enzyme activities in rice-koji.2021

    • Author(s)
      Taku Matsumoto, Kouji Kojima, Noriaki Saigusa, Yuji Teramoto
    • Journal Title

      International Journal of Biomass and Renewables

      Volume: 10 Pages: 18-24

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] High-Light-Induced Stress Activates Lipid Deacylation at the sn-2 Position in the Cyanobacterium Synechocystis Sp. PCC 68032021

    • Author(s)
      Kojima Kouji、Matsumoto Ui、Keta Sumie、Nakahigashi Kenji、Ikeda Kazutaka、Takatani Nobuyuki、Omata Tatsuo、Aichi Makiko
    • Journal Title

      Plant and Cell Physiology

      Volume: 63 Issue: 1 Pages: 82-91

    • DOI

      10.1093/pcp/pcab147

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 食品におけるイソプレノイド化合物の役割2020

    • Author(s)
      小島幸治
    • Journal Title

      BIO九州

      Volume: 228 Pages: 7-12

    • NAID

      40022349104

    • Related Report
      2020 Research-status Report
  • [Journal Article] Two nucleolar proteins, GDP1 and OLI2, function as ribosome biogenesis factors and are preferentially involved in promotion of leaf cell proliferation without strongly affecting leaf adaxial-abaxial patterning in Arabidopsis thaliana.2018

    • Author(s)
      Kojima K, Tamura J, Chiba H, Fukuda K, Tsukaya H, Horiguchi G
    • Journal Title

      Frontiers in Plant Science

      Volume: ー Pages: 2240-2240

    • DOI

      10.3389/fpls.2017.02240

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Plastid gene expression is essential for lateral root stem-cell patterning, symplastic connectivity, and root plastid distribution in Arabidopsis thaliana2018

    • Author(s)
      Nakata MT, Sato M, Wakazaki M, Sato N, Kojima K Sekine A, Nakamura S, Shikanai T, Toyooka K, Tsukaya H, Horiguchi G
    • Journal Title

      Biology Open

      Volume: 7 Pages: 028175-028175

    • DOI

      10.1242/bio.028175

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Aspergillus oryzae RIB40株で製麹した米麹のグルコアミラーゼ活性に及ぼす音波照射の影響2022

    • Author(s)
      松本 拓,小島 幸治,三枝 敬明,寺本 祐司
    • Organizer
      日本農芸化学会 2021年度大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] サポジラ(Manilkara zapota)由来 trans型プレニルトランスフェラーゼの酵素機能解析2021

    • Author(s)
      三輪 幸祐,廣森 美樹,青木 裕一,和氣 駿之,小島 幸治,山下 哲,山口 晴彦,宮城 ゆき乃,戸澤 譲,中山 亨,高橋 征司
    • Organizer
      第31回イソプレノイド研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Aspergillus属の分生子に対する音波照射と米麹の酵素活性への影響2021

    • Author(s)
      松本 拓,小島 幸治,三枝 敬明,寺本 祐司
    • Organizer
      日本農芸化学会 2021年度西日本・中四国・関西支部 合同大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] サポジラ(Manilkara zapota)由来trans型ポリイソプレノイド 生合成酵素のクローニングと機能解析2020

    • Author(s)
      三輪幸祐,廣森美樹,青木裕一,和氣駿之,小島幸治,山下哲,山口晴彦,宮城ゆき乃,戸澤譲,中山亨,高橋征司
    • Organizer
      第30回 イソプレノイド研究会
    • Related Report
      2020 Research-status Report
  • [Presentation] パラゴムノキの天然ゴム生合成関連タンパク質の機能解析2019

    • Author(s)
      小島幸治、山口真琴、廣森美樹、和氣駿之、山下哲、戸澤譲、山口晴彦、宮城ゆき乃、伏原和久、中山亨、高橋征司
    • Organizer
      第2回天然ゴム研究会(理研、鶴見)
    • Related Report
      2019 Research-status Report
  • [Presentation] Interaction and its functional correlation of factors constituting the biosynthetic machinery of natural rubber from Hevea brasiliensis2019

    • Author(s)
      Kouji Kojima, Makoto Yamaguchi, Tomoki Ishii, Miki Suenaga-Hiromori, Toshiyuki Waki, Satoshi Yamashita, Yuzuru Tozawa, Haruhiko Yamaguchi, Yukino Miyagi-Inoue, Kazuhisa Fushihara, Toru Nakayama, Seiji Takahashi
    • Organizer
      Japan-Taiwan Plant Biology 2019 (JTPB2019)、Nagoya Univ. Nagoya, Japan
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] In vitro natural rubber biosynthesis by prenyltransferases introduced on rubber particles from Hevea brasiliensis2018

    • Author(s)
      Tomoki Ishii, Miki Suenaga-Hiromiri, Satoshi Yamashita, Fumihiro Yanbe, Toshiyuki Waki, Kouji Kojima, Haruhiko Yamaguchi, Yukino Miyagi-Inoue, Kazuhisa Fushihara
    • Organizer
      23th International Symposium on Plant Lipids (ISPL2018)、Osanbashi-Hall, Yokohama, Japan
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Production of natural rubber in vitro from reconstituted-rubber synthase complex on rubber particles with Escherichia coli cell-free translation system2018

    • Author(s)
      Kouji Kojima,Miki Suenaga-Hiromori, Satoshi Yamashita,Yuzuru Tozawa, Haruhiko Yamaguchi,Yukino Miyagi-Inoue,Kazuhisa Fushihara,Toru Nakayama, Seiji Takahashi
    • Organizer
      23th International Symposium on Plant Lipids (ISPL2018)、Osanbashi-Hall, Yokohama, Japan
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] 天然ゴム生合成機構におけるゴム粒子の重要性2018

    • Author(s)
      石井 智樹,末永 美樹,山下 哲,山家 史大,和氣 駿之,小島 幸治, 山口 晴彦,井之上 ゆき乃,伏原 和久,中山 亨,高橋 征司
    • Organizer
      農芸化学会2018年度名古屋大会(名城大学、名古屋)
    • Related Report
      2018 Research-status Report
  • [Presentation] 大腸菌無細胞翻訳系を用いた天然ゴム合成酵素の再構成2018

    • Author(s)
      小島 幸治、山下 哲、戸澤 譲、山口 晴彦、井之上 ゆき乃、伏原 和久、 中山 亨、高橋 征司
    • Organizer
      第59回日本植物生理学会年会(札幌コンベンションセンター、札幌)
    • Related Report
      2018 Research-status Report
  • [Presentation] パラゴムノキの天然ゴム合成反応におけるゴム粒子の役割2018

    • Author(s)
      小島 幸治、廣森 美樹、山家 史大、石井 智樹、和氣 駿之、山下 哲、 戸澤 譲、山口 晴彦、井之上 ゆき乃、伏原 和久、中山 亨、高橋 征司
    • Organizer
      日本農芸化学会 東北・北海道支部合同大会(東北支部第153回大会、東北大学、仙台)
    • Related Report
      2018 Research-status Report
  • [Presentation] パラゴムノキの天然ゴム生合成マシナリを構成する因子の相互作用と機能相関2018

    • Author(s)
      小島 幸治、山口 真琴、石井 智樹、廣森 美樹、和氣 駿之、山下 哲、戸澤 譲、 山口 春彦、井之上 ゆき乃、伏原 和久、中山 亨、高橋 征司
    • Organizer
      第60回日本植物生理学会年会(名古屋大学、名古屋)
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2023-01-30  

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