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Trypsin-immobilized nanosheets for increased proteolytic stability

Research Project

Project/Area Number 25410230
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Polymer/Textile materials
Research InstitutionKumamoto University

Principal Investigator

Sakata Masayo  熊本大学, 自然科学研究科, 准教授 (60187391)

Co-Investigator(Renkei-kenkyūsha) TANIGUCHI TAKAAKI  熊本大学, 大学院自然科学研究科, 助教 (50583415)
Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2013: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords酸化グラフェン / ナノシート / トリプシン / 固定化酵素 / 酵素活性 / 自己消化 / 酵素安定性 / 酸化グラフェンナノシート / プロテアーゼ / ナノ材料 / 酵素担体 / 磁性粒子
Outline of Final Research Achievements

In this paper, we report a simple and natural method for the preparation of enzyme-immobilized grapheme oxide nanosheets (GO sheets) by the diimide-activated amidation of GO-bound carboxylic acids. Trypsin was used as a model enzyme for the investigation. In order to study the actual stabilization of immobilized trypsin, we also compared the thermal and chemical stabilities of the immobilized trypsin with the free-trypsin stabilities under various conditions. The trypsin-immobilized GO sheets (1.9 mg per mg-GO sheets) showed significantly higher thermal and chemical stabilities than the free form. After heat treatment at 50 °C for 6 h, the trypsin-GO sheets retained 80% of its initial activity, but free trypsin only retained 4% of its initial activity.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (21 results)

All 2015 2014 2013 Other

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Open Access: 2 results) Presentation (12 results) (of which Int'l Joint Research: 3 results,  Invited: 1 results) Book (1 results) Remarks (2 results)

  • [Journal Article] γ-Cyclodextrin/polyurethane Copolymer Adsorbent for Selective Removal of Endotoxin from DNA2015

    • Author(s)
      Masayo Sakata, Kasane Kimura, Koji Uezono, Masami Todokoro
    • Journal Title

      Journal of the Society of Japanese Women Scientists

      Volume: 15 Pages: 28-32

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Preparation of crooss-linked γ-cyclodextrin particles for selective removal endotoxin2015

    • Author(s)
      Masayo Sakata
    • Journal Title

      Cellulose Communications

      Volume: 22 Pages: 191-195

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] γ-Cyclodextrin/polyurethane Copolymer Adsorbent for Selective Removal of Endotoxin from DNA Solution2015

    • Author(s)
      Masayo Sakata, Kasane Kimura, Koji Uezono, Masami Todokoro
    • Journal Title

      Journal of the Society of Japanese Women Scientists

      Volume: 15 Pages: 28-32

    • NAID

      130005063871

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Polymeric γ-Cyclodextrin Adsorbent for Selective Removal of Endotoxin from DNA Solution2014

    • Author(s)
      坂田眞砂代, 戸所正美
    • Journal Title

      KOBUNSHI RONBUNSHU

      Volume: 71 Issue: 7 Pages: 283-292

    • DOI

      10.1295/koron.71.283

    • NAID

      130004793164

    • ISSN
      0386-2186, 1881-5685
    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] γ-Cyclodextrin-polyurethane copolymer adsorbent for selective removal of endotoxin from DNA solution2013

    • Author(s)
      Masayo Sakata, Koji Uezono, Kasane Kimura, Masami Todokoro
    • Journal Title

      Analytical Biochemistry

      Volume: 443 Issue: 1 Pages: 41-45

    • DOI

      10.1016/j.ab.2013.08.010

    • NAID

      130005063871

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Surface initiated ATRP: synthesis and characterization of functional polymers grafted on modified cellulose beads2013

    • Author(s)
      Md. Ashaduzzaman, Kei Ishikura, Masayo Sakata, Masashi Kunitake
    • Journal Title

      International Letters of Chemistry, Physics and Astronomy

      Volume: 8 Pages: 243-248

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Presentation] Masayo Sakata, Kasane Kimura, Taku Matsuo, Masami Todokoro2015

    • Author(s)
      Removal of endotoxin from DNA solution by cyclodextrin/urethane copolymer
    • Organizer
      The 2015 International Chemical Congress of Pacific Basin Societies
    • Place of Presentation
      Honolulu, Hawaii, U.S.A
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Immobolization of glucoamylase on cellulose supports by covalent binding for increased saccharification stability2015

    • Author(s)
      Towako Sakamoto, Ryosuke Harada, Masayo Sakata
    • Organizer
      The 2015 International Chemical Congress of Pacific Basin Societies
    • Place of Presentation
      Honolulu, Hawaii, U.S.A
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Aminated cellulose nano-fibers for removal of endotoxin from protein solution2015

    • Author(s)
      47.Risa Harada, Towako Sakamot1, Megumi Shimizu, Daisuke Nakamura, Yuki Maed, Masayo Sakata
    • Organizer
      2015 Pusan-Gyeongnam/Kyushu-Seibu Joint Symposium on High Polymer (17th) and Fiber (15th)
    • Place of Presentation
      Dong-A University (Bumin Campus), Busan, Korea
    • Year and Date
      2015-11-12
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Cyclodextrin polymer adsorbents for removal of endotoxin from bio-products2014

    • Author(s)
      M. Sakata, K. Kimura, T. Matsuo, M. Todokoro
    • Organizer
      The 10th SPSJ International Polymer Coference
    • Place of Presentation
      つくば国際会議場
    • Year and Date
      2014-12-02 – 2014-12-05
    • Related Report
      2014 Research-status Report
  • [Presentation] Chromatographic separation of DNA from bio-product solution by cellulose beads grafted with cationic linear polymer2014

    • Author(s)
      E. Ikegami, Y. Goto, M. Sakata
    • Organizer
      The 10th SPSJ International Polymer Coference
    • Place of Presentation
      つくば国際会議場
    • Year and Date
      2014-12-02 – 2014-12-05
    • Related Report
      2014 Research-status Report
  • [Presentation] トリプシン固定化酸化グラフェンナノシートの開発-トリプシン化学修飾法2014

    • Author(s)
      原田諒祐、園田将平、坂田眞砂代
    • Organizer
      第63回高分子討論会
    • Place of Presentation
      長崎大学
    • Year and Date
      2014-09-24 – 2014-09-26
    • Related Report
      2014 Research-status Report
  • [Presentation] 包接能を利用するエンドトキシン選択分離剤の開発 (ウレタン架橋シクロデキストリン微粒子)2014

    • Author(s)
      坂田眞砂代 、木村かさね、松尾拓、戸所正美
    • Organizer
      第63回高分子討論会
    • Place of Presentation
      長崎大学
    • Year and Date
      2014-09-24 – 2014-09-26
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] LPS選択除去のための酸化グラフェン固定化セルロースビーズ/γ-シクロデキストリン  -ハイブリッド吸着剤の開発2014

    • Author(s)
      松尾拓、木村かさね、坂田眞砂代
    • Organizer
      第63回高分子討論会
    • Place of Presentation
      長崎大学
    • Year and Date
      2014-09-24 – 2014-09-26
    • Related Report
      2014 Research-status Report
  • [Presentation] Preparation of trypsin-immobilized graphene oxide nanosheets and their application for increased proteolytic stability

    • Author(s)
      Masayo Sakata, Shohei Sonoda, Asami Funatsu, Takaaki Taniguchi, Yasumichi Matsumoto
    • Organizer
      5th International Conference on Recent Progress in Graphene Research
    • Place of Presentation
      Tokyo Institute of Technology, Oookayama,Tokyo
    • Related Report
      2013 Research-status Report
  • [Presentation] Preparation of magnetic graphene oxide nanosheets for enzyme immobilization support

    • Author(s)
      Shohei Sonoda, Aki Kuwahara, Asami Funatsu, Takaaki Taniguchi, Yasumichi Matsumoto, Masayo Sakata
    • Organizer
      5th International Conference on Recent Progress in Graphene Research
    • Place of Presentation
      Tokyo Institute of Technology, Oookayama,Tokyo
    • Related Report
      2013 Research-status Report
  • [Presentation] Preparation of cellulose beads grafted with cationic polymer chains through ATRT for separation of DNA

    • Author(s)
      Masayo Sakata, Eri Ikegami , Yusuke Goto
    • Organizer
      40th International Symposium on High Performance Liquid Phase Separations and Related Techniques
    • Place of Presentation
      The Hotel Grand Chancellor Hotel, Hobart, Tasmania, Australia
    • Related Report
      2013 Research-status Report
  • [Presentation] Preparation of magnetic graphene oxide nanosheets for enzyme immobilization support

    • Author(s)
      Masayo Sakata, Shohei Sonoda, Asami Funatsu, Ryosuke Harada
    • Organizer
      Ist International Sympsium on Graphene Oxide
    • Place of Presentation
      Kumamoto Univ., Kumamoto, Japan
    • Related Report
      2013 Research-status Report
  • [Book] 吸着・分離材料の設計、性能評価と新しい応用2015

    • Author(s)
      坂田眞砂代
    • Total Pages
      8
    • Publisher
      技術情報協会
    • Related Report
      2015 Annual Research Report
  • [Remarks] 熊本大学工学部國武研究室

    • URL

      http://chem.chem.kumamoto-u.ac.jp/~polymers/

    • Related Report
      2015 Annual Research Report 2014 Research-status Report
  • [Remarks] 熊本大学大学院自然科学研究科國武研究室ホームページ

    • URL

      http://chem.chem.kumamoto-u.ac.jp/~polymers/index.html

    • Related Report
      2013 Research-status Report

URL: 

Published: 2014-07-25   Modified: 2019-07-29  

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