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Development of Enzyme Immobilization technique to Improve Stability and Its Application for Microreactor

Research Project

Project/Area Number 20310074
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Microdevices/Nanodevices
Research InstitutionKyushu University

Principal Investigator

MIYAZAKI Masaya  Kyushu University, 生産計測技術研究センター, 主任研究員 (70344231)

Project Period (FY) 2008 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥19,370,000 (Direct Cost: ¥14,900,000、Indirect Cost: ¥4,470,000)
Fiscal Year 2010: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2009: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2008: ¥10,270,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥2,370,000)
Keywordsマイクロ・ナノ化学 / 酵素反応 / マイクロリアクター / 固定化触媒 / 安定化
Research Abstract

In this research, we developed novel cross-linking immobilization techniques by using cross-linking of complex of synthetic polymers and enzymes, based on our original immobilization method. By this method, we could immobilize enzymes onto the microchannel wall, with improved stability against denaturating conditions. We also applied this technique to develop the micro enzyme process.

Report

(4 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report
  • 2008 Annual Research Report
  • Research Products

    (51 results)

All 2011 2010 2009 2008 Other

All Journal Article (19 results) (of which Peer Reviewed: 11 results) Presentation (29 results) Book (2 results) Remarks (1 results)

  • [Journal Article] A simple and rapid analysis of disulfide bonds in proteins using thermostable protease-immobilized microfluidic reactors integrated with MS2011

    • Author(s)
      Yamaguchi, H., Miyazaki, M., Maeda, H.
    • Journal Title

      Peptide Science 2010

      Volume: (掲載確定)

    • NAID

      10028238220

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Proteolysis approach without chemical modification for a simple and rapid analysis of disulfide bonds using thermostable protease-immobilized microreactors2010

    • Author(s)
      山口浩、宮崎真佐也、前田英明
    • Journal Title

      Proteomics 10巻

      Pages: 2942-2949

    • Related Report
      2010 Final Research Report
  • [Journal Article] Multidigestion in continuous flow tandem protease-immobilized microreactors for proteomic analysis2010

    • Author(s)
      山口浩、宮崎真佐也、河済博文、前田英明
    • Journal Title

      Analytical Biochemistry 407巻

      Pages: 12-18

    • Related Report
      2010 Final Research Report
  • [Journal Article] Refolding of difficult-to-fold proteins by a gradual decrease of denaturant using microfluidic chips2010

    • Author(s)
      Yamaguchi, H., Miyazaki, M., Briones-Nagata, M.P., Maeda, H.
    • Journal Title

      J.Biochem.

      Volume: 147 Pages: 895-903

    • NAID

      10027876657

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Multidigestion in continuous flow tandem protease-immobilized mirror eactors for proteomic analysis2010

    • Author(s)
      Yamaguchi, H., Miyazaki, M., Kawazumi, H., Maeda, H.
    • Journal Title

      Anal.Biochem.

      Volume: 407 Pages: 12-18

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Proteolysis approach without chemical modification for a simple and rapid analysis of disulfide bonds using thermostable protease-immobilized microreactors.2010

    • Author(s)
      Yamaguchi, H., Miyazaki, M., Maeda, H.
    • Journal Title

      Proteomics

      Volume: 10 Pages: 2942-2949

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid post-translational modification analysis by enzyme-immobilized microreactor2010

    • Author(s)
      Hiroshi Yamaguchi, ほか
    • Journal Title

      Peptide Science 2009

      Pages: 451-454

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Refolding of Difficult-to-fold Proteins by a Gradual Decrease of Denaturant Using Microfluidic Chips2010

    • Author(s)
      Hiroshi Yamaguchi, ほか
    • Journal Title

      J. Biochem. (印刷中)

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] マイクロ空間を用いる有機合成反応2009

    • Author(s)
      宮崎真佐也、前田英明
    • Journal Title

      化学と教育 57巻

      Pages: 364-367

    • Related Report
      2010 Final Research Report
  • [Journal Article] Rapid and efficient proteolysis for proteomic analysis by protease-immobilized microreactor2009

    • Author(s)
      山口浩、宮崎真佐也, ほか
    • Journal Title

      Electrophoresis 30巻

      Pages: 3257-3264

    • Related Report
      2010 Final Research Report
  • [Journal Article] Polymer Chemistry in Microfluidic Reactor2009

    • Author(s)
      宮崎真佐也, ほか
    • Journal Title

      Micro and Nanosystem 1巻

      Pages: 193-204

    • Related Report
      2010 Final Research Report
  • [Journal Article] Rapid and efficient proteolysis for proteomic analysis by protease-immobilized microreactor2009

    • Author(s)
      Hiroshi Yamaguchi, ほか
    • Journal Title

      Electrophoresis 30

      Pages: 3257-3264

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] マイクロ空間を用いる有機合成反応2009

    • Author(s)
      宮崎真佐也、前田英明
    • Journal Title

      化学と教育 57

      Pages: 364-367

    • Related Report
      2009 Annual Research Report
  • [Journal Article] Polymer Chemistry in Microfluidic Reactor2009

    • Author(s)
      Masaya Miyazaki, ほか
    • Journal Title

      Micro and Nanosystem 1

      Pages: 193-204

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid Protein Digestion by Protease-immobilized Microreactor2009

    • Author(s)
      Hiroshi Yamaguchi, 他
    • Journal Title

      Peptide Science 2008

      Pages: 519-520

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Enzymatic processing in microfluidic reactors2008

    • Author(s)
      宮崎真佐也, ほか
    • Journal Title

      Enzymatic processing in microfluidic reactors 25巻

      Pages: 405-428

    • Related Report
      2010 Final Research Report
  • [Journal Article] Biomolecular Synthesis in Microfluid2008

    • Author(s)
      宮崎真佐也、前田英
    • Journal Title

      Encyclopedia of Micro-and Nanofluidics

      Pages: 76-84

    • Related Report
      2010 Final Research Report
  • [Journal Article] Enzymatic processing in microfluidic reactors.2008

    • Author(s)
      Masaya Miyazaki, 他
    • Journal Title

      Biotechnology and Genetic Engineering Reviews 25

      Pages: 405-428

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Biomolecular Synthesis in Microfluid2008

    • Author(s)
      Masaya Miyazaki, Hideaki Maeda
    • Journal Title

      Encyclopedia of Micro- and Nanofluidics

      Pages: 76-84

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Presentation] 合成プローブを用いた光架橋とマイクロリアクタを用いた配列解析による分子間相互作用解析技術の開発2011

    • Author(s)
      宮崎真佐也、山口浩、犬養吉成、長田聡史、兒玉浩明
    • Organizer
      第13回佐賀大学連携大学院産学官交流セミナー
    • Place of Presentation
      佐賀市
    • Year and Date
      2011-01-27
    • Related Report
      2010 Annual Research Report
  • [Presentation] Simple and rapid proteolysis by protease-immobilized microfluidic reactor for proteomic analysis2010

    • Author(s)
      宮崎真佐也、山口浩、前田英明
    • Organizer
      2010 Interantional Chemical Congress of Pacific Basin Societies(Pacifichem 2010)
    • Place of Presentation
      Honolulu, HI, USA
    • Year and Date
      2010-12-16
    • Related Report
      2010 Annual Research Report
  • [Presentation] Proteolysis approach without chemical modification for a simple and rapid analysis of disulfide bonds using thermostable protease-immobilized microreactors.2010

    • Author(s)
      宮崎真佐也、山口浩、前田英明
    • Organizer
      第33回日本分子生物学会年会・第83回日本生化学会大会合同大会(BMB2010)
    • Place of Presentation
      神戸市
    • Year and Date
      2010-12-08
    • Related Report
      2010 Annual Research Report
  • [Presentation] A simple and rapid analysis of disulfide bonds in proteins using ther mostable protease-immobilized microfluidic reactors integrated with MS2010

    • Author(s)
      宮崎真佐也、山口浩、前田英明
    • Organizer
      The 5^<th> International Peptide Symposium
    • Place of Presentation
      京都市
    • Year and Date
      2010-12-08
    • Related Report
      2010 Annual Research Report
  • [Presentation] マイクロ流体デバイスを用いたプロテオミクス解析2010

    • Author(s)
      宮崎真佐也
    • Organizer
      九大先導研ナノバイオ・JST先端計測合同シンポジウム
    • Place of Presentation
      福岡市(招待講演)
    • Year and Date
      2010-10-25
    • Related Report
      2010 Annual Research Report
  • [Presentation] 酵素固定化マイクロリアクターを用いたタンパク質翻訳後修飾の迅速解析2010

    • Author(s)
      吉塚紗央里、山口浩、宮崎真佐也、前田英明
    • Organizer
      第47回化学関連支部合同九州大会
    • Place of Presentation
      北九州市
    • Year and Date
      2010-07-10
    • Related Report
      2010 Annual Research Report
  • [Presentation] プロテアーゼ固定化マイクロリアクターによるジスルフィド結合の迅速解析2010

    • Author(s)
      宮崎真佐也、山口浩、前田英明
    • Organizer
      平成22年度日本生化学会九州支部例会
    • Place of Presentation
      鹿児島市
    • Year and Date
      2010-05-23
    • Related Report
      2010 Annual Research Report
  • [Presentation] Enzymatic Conversion in Enzyme-Immobilized Microchannel Reactor with Cross-Linked Enzyme Membrane2010

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      11th International Conference on Microreaclion Technology(IMRET11)
    • Place of Presentation
      京都市
    • Year and Date
      2010-03-08
    • Related Report
      2009 Annual Research Report
  • [Presentation] プロテオミクス解析用マイクロリアクターの開発2010

    • Author(s)
      山口浩, ほか
    • Organizer
      第12回連携大学院産学官交流セミナー
    • Place of Presentation
      鳥栖市
    • Year and Date
      2010-01-18
    • Related Report
      2009 Annual Research Report
  • [Presentation] マイクロ空間化学のバイオテクノロジーへの応用2010

    • Author(s)
      宮崎真佐也
    • Organizer
      第4回ナノバイオデバイス・ワークショップ
    • Place of Presentation
      つくば市
    • Year and Date
      2010-01-15
    • Related Report
      2009 Annual Research Report
  • [Presentation] マイクロ空間化学のバイオテクノロジーへの応用2010

    • Author(s)
      宮崎真佐也
    • Organizer
      第4回ナノバイオデバイスワークショップ
    • Place of Presentation
      つくば市
    • Year and Date
      2010-01-11
    • Related Report
      2010 Final Research Report
  • [Presentation] Simple and rapid proteolysis by protease-immobilized microfluidic reactor for proteomic analysis2010

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      2010 International Chemical Congress of Pacific Basin Societies (Pacifichem2010)
    • Place of Presentation
      Honolulu, HI, U.S.A.
    • Related Report
      2010 Final Research Report
  • [Presentation] A simple and rapid analysis of disulfide bonds in proteins using thermostable proteaseimmobilized microfluidic reactors integrated with MS2010

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      The 5th International Peptide Symposium
    • Place of Presentation
      京都市
    • Related Report
      2010 Final Research Report
  • [Presentation] マイクロ流体デバイスを用いたプロテオミクス解析2010

    • Author(s)
      宮崎真佐也
    • Organizer
      九州大学先導物質化学研究所・JST 先端計測合同シンポジウム
    • Place of Presentation
      福岡市
    • Related Report
      2010 Final Research Report
  • [Presentation] Enzymatic Conversion in Enzyme-Immobilized Microchannel Reactor with Cross-Linked Enzyme Membrane2010

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      11th International Conference on Microreaction Technology (IMRET11)
    • Place of Presentation
      京都市
    • Related Report
      2010 Final Research Report
  • [Presentation] Rapid Post-Translational Modification Analysis by Enzyme- Immobilized Microreactor2009

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      International Symposium on Microchemistry and Microsystems(ISMM2009)
    • Place of Presentation
      金沢市
    • Year and Date
      2009-11-07
    • Related Report
      2009 Annual Research Report
  • [Presentation] 酵素固定化マイクロリアクターによるタンパク質翻訳後修飾の迅速分析2009

    • Author(s)
      山口浩, ほか
    • Organizer
      第46回ペプチド討論会
    • Place of Presentation
      北九州市
    • Year and Date
      2009-11-05
    • Related Report
      2009 Annual Research Report
  • [Presentation] 酵素固定化マイクロ流体デバイスによるタンパク質リン酸化の迅速解析2009

    • Author(s)
      山口浩, ほか
    • Organizer
      第82回日本生化学会大会
    • Place of Presentation
      神戸市
    • Year and Date
      2009-10-02
    • Related Report
      2009 Annual Research Report
  • [Presentation] PEG化ポリリジンによる酵素の固定化と活性への効果2009

    • Author(s)
      山口浩, ほか
    • Organizer
      第46回化学関連支部合同九州大会
    • Place of Presentation
      北九州市
    • Year and Date
      2009-07-11
    • Related Report
      2009 Annual Research Report
  • [Presentation] 酵素固定化マイクロリアクターを用いたタンパク質の迅速加水分解2009

    • Author(s)
      山口浩, ほか
    • Organizer
      第46回化学関連支部合同九州大会
    • Place of Presentation
      北九州市
    • Year and Date
      2009-07-11
    • Related Report
      2009 Annual Research Report
  • [Presentation] タンパク質の迅速加水分解反応を行うマイクロリアクターの開発2009

    • Author(s)
      山口浩, ほか
    • Organizer
      第19回化学とマイクロ・ナノシステム研究会
    • Place of Presentation
      東広島市
    • Year and Date
      2009-05-24
    • Related Report
      2009 Annual Research Report
  • [Presentation] プロテアーゼ固定化マイクロリアクターを用いたタンパク質の迅速加水分解2009

    • Author(s)
      山口浩, ほか
    • Organizer
      平成21年度 日本生化学会九州支部例会
    • Place of Presentation
      福岡市
    • Year and Date
      2009-05-17
    • Related Report
      2009 Annual Research Report
  • [Presentation] Rapid Post-Translational Modification Analysis by Enzyme-Immobilized Microreactor2009

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      International Synposium on Microchemistry and Microsystems (ISMM2009)
    • Place of Presentation
      金沢市
    • Related Report
      2010 Final Research Report
  • [Presentation] Cross-linked Aggregated Hydrogel Membrane Formation as Immobilization Method of Enzyme Onto Microchannel Surface2008

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      The IUMRS International Conference in Asia (IUMRS-ICA 2008)
    • Place of Presentation
      名古屋市
    • Related Report
      2010 Final Research Report
  • [Presentation] Protease-immobilized microreactor for rapid protein digestion2008

    • Author(s)
      宮崎真佐也, ほか
    • Organizer
      17th Meeting of Methods in Protein Structure Analysis (MPSA2008)
    • Place of Presentation
      札幌市
    • Related Report
      2010 Final Research Report 2008 Annual Research Report
  • [Presentation] Cross-Linked Aggregated Hydrogel Membrane Formation as Immobilization Method of Enzymes onto Microchannel Surface2008

    • Author(s)
      Masaya Miyazaki, 他
    • Organizer
      The IUMRS International Conference in Asia(IUMRS-ICA2008)
    • Place of Presentation
      名古屋市
    • Related Report
      2008 Annual Research Report
  • [Presentation] 酵素固定化マイクロリアクターにおけるPEGイヒポリリジンの効果2008

    • Author(s)
      山口浩, 他
    • Organizer
      第18回化学とマイクロ・ナノシステム研究会
    • Place of Presentation
      京都市
    • Related Report
      2008 Annual Research Report
  • [Presentation] Protease-immobilized microreactor for rapid protein digestion2008

    • Author(s)
      角久間隆文, 他
    • Organizer
      第18回化学とマイクロ・ナノシステム研究会
    • Place of Presentation
      京都市
    • Related Report
      2008 Annual Research Report
  • [Presentation] 酵素固定化マイクロリアクターによるタンパク質の迅速加水分解2008

    • Author(s)
      山口浩, 他
    • Organizer
      第45回ぺプチド討論会
    • Place of Presentation
      東京都
    • Related Report
      2008 Annual Research Report
  • [Book] Limited proteolysis in proteomics using protease-immobilized microreactors.Method in Molecular Biology Series-Functional Genomics, 2nd ed., (Kaufmann, M.and Klinger, C.eds)2011

    • Author(s)
      Yamaguchi, H., Miyazaki, M., Maeda, H.
    • Publisher
      Humana Press(Springer reference)(掲載確定)
    • Related Report
      2010 Annual Research Report
  • [Book] Limited proteolysis in proteomics using protease-immobilized microreactors.、Method in Molecular Biology Series-Functional Genomics, 2nd ed., (Kaufmann, M. and Klinger, C. eds)

    • Author(s)
      山口浩、宮崎真佐也、前田英明
    • Publisher
      Humana Press (受理済)
    • Related Report
      2010 Final Research Report
  • [Remarks] ホームページ

    • URL

      http://unit.aist.go.jp/msrc/ci/index.html

    • Related Report
      2010 Final Research Report

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Published: 2008-04-01   Modified: 2016-04-21  

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