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Cell-free approach to design cellulose nanofibers

Research Project

Project/Area Number 18K06142
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43030:Functional biochemistry-related
Research InstitutionOkinawa Institute of Science and Technology Graduate University

Principal Investigator

Nakashima Keisuke  沖縄科学技術大学院大学, マリンゲノミックスユニット, スタッフサイエンティスト (10422924)

Co-Investigator(Kenkyū-buntansha) 今井 友也  京都大学, 生存圏研究所, 教授 (90509142)
Project Period (FY) 2018-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywords無細胞タンパク質合成系 / 膜タンパク質 / セルロース合成酵素 / セルロース / インビトロ合成 / ホヤ / カタユウレイボヤ / 無細胞発現 / 被嚢動物 / 無細胞タンパク発現 / 結晶繊維 / 結晶多形
Outline of Final Research Achievements

This study aimed to produce cellulose crystal fibers with desired properties (on-demand artificial synthesis). First, a cell-free reaction system was established to synthesize ascidian cellulose synthase with a molecular weight of over 200 kDa using insect cell extracts. The synthesized enzyme, which is a membrane protein, aggregates and becomes inactive upon synthesis if there is no suitable hydrophobic environment in the vicinity. Therefore, we incorporated various artificial hydrophobic domains into the reaction system and found conditions under which 80% of the cell-free synthesis products are incorporated into the phospholipid environment. We set up a system for sensitive, rapid, and quantitative evaluation of substrate uptake activity using radioisotopes, and investigated optimal conditions for synthesis. The reaction products were analyzed by electron microscopy, infrared spectroscopy, and electron diffraction, and the synthesis of cellulose crystal fibers was confirmed.

Academic Significance and Societal Importance of the Research Achievements

従来、植物由来のセルロースが木材・パルプ繊維として利用されることが主であったが、近年はそれを結晶繊維のレベルで新たな機能を発現させた高付加価値の新資源として利用する試みがなされている。天然物であるセルロースの結晶繊維は、物性を左右する諸性質(太さ・長さ・単位胞組成など)が不均一であり、それが生合成の過程でどのように制御されているかは不明である。そのしくみを解明し、活用することで、新たなセルロース利用が可能になる。本研究は、自然界で唯一単位胞レベルで均質な結晶繊維をつくる生物であるホヤのセルロース合成酵素に着目し、望みの性状の結晶繊維を創ること(セルロースのオンデマンド人工合成)を目指した。

Report

(5 results)
  • 2022 Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (5 results)

All 2020 2019 2018

All Journal Article (4 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 3 results,  Open Access: 3 results) Presentation (1 results)

  • [Journal Article] Phylogenetic Analyses of Glycosyl Hydrolase Family 6 Genes in Tunicates: Possible Horizontal Transfer2020

    • Author(s)
      Kun-Lung Li, Keisuke Nakashima, Jun Inoue, and Noriyuki Satoh
    • Journal Title

      Genes

      Volume: 11 Issue: 8 Pages: 937-937

    • DOI

      10.3390/genes11080937

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] ORTHOSCOPE Analysis Reveals the Presence of the Cellulose Synthase Gene in All Tunicate Genomes but Not in Other Animal Genomes2019

    • Author(s)
      Jun Inoue, Keisuke Nakashima, and Noriyuki Satoh
    • Journal Title

      Genes

      Volume: 10 Issue: 4 Pages: 1-9

    • DOI

      10.3390/genes10040294

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] 腸内細菌による粘液層への定着は動物進化上の新しい特徴である2019

    • Author(s)
      中島啓介、菊池さくら
    • Journal Title

      実験医学

      Volume: 37 Pages: 69-72

    • Related Report
      2018 Research-status Report
  • [Journal Article] A comparative approach to decipher intestinal animal-microbe associations2018

    • Author(s)
      Keisuke Nakashima
    • Journal Title

      Microbial Cell

      Volume: 5 Issue: 11 Pages: 522-524

    • DOI

      10.15698/mic2018.11.658

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] 脊椎動物の消化管におけるキチン製バリア免疫機構の発見とその喪失が哺乳類の腸内細菌に及ぼした影響2018

    • Author(s)
      中島啓介、菊池さくら、佐藤矩行、小川悠、游智凱、相馬智史、渡邊壮一、金子豊二、木村聡
    • Organizer
      セルロース学会
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2024-01-30  

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