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2013 Fiscal Year Final Research Report

Research and development of the highly-active cathodic enzyme necessary for the practical use of the bio-fuel cell

Research Project

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Project/Area Number 23580131
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Applied biochemistry
Research InstitutionKanazawa University

Principal Investigator

KATAOKA KUNISHIGE  金沢大学, 物質化学系, 教授 (40252712)

Co-Investigator(Renkei-kenkyūsha) KANO Kenji  京都大学, 大学院農学研究科, 教授 (10152828)
HIGUCHI Yoshiki  兵庫県立大学, 大学院生命理学研究科, 教 授 (90183574)
Project Period (FY) 2011 – 2013
Keywordsバイオ燃料電池 / マルチ銅オキシダーゼ / ラッカーゼ / 酵素電極 / 機能改変
Research Abstract

For the development of the platinum-substitute anodic enzyme catalysts for bio-fuel cell, I performed the in silico screening to find new multi-copper oxidases and the mutations against the common enzymes to improve their oxygen reduction activity. In the screening for the new multi-copper oxidases, I found two laccase genes from an anaerobic bacterium and an insect. The heterologous expression systems for the laccase genes were successfully constructed, and the recombinant laccase from anaerobe was found to have unreported properties so far. Furthermore, the improvements of the bacterial common enzymes in the oxygen-reduction activity and the electrode response were achieved by protein engineering.

  • Research Products

    (23 results)

All 2014 2013 2012 2011

All Journal Article (7 results) (of which Peer Reviewed: 7 results) Presentation (15 results) Book (1 results)

  • [Journal Article] New Insights into the Catalytic Active-Site Structure of Multicopper Oxidases2014

    • Author(s)
      H. Komori, R. Sugiyama, K. Kataoka, K. Miyazaki, Y. Higuchi, and T. Sakurai
    • Journal Title

      Acta Cryst

      Volume: D70 Pages: 772-779

    • DOI

      10.1107/S1399004713033051

    • Peer Reviewed
  • [Journal Article] Crystal Structure of the CueO Mutants at Glu506, the Key Amino Acid Located in the Proton Transfer Pathway for Dioxygen Reduction2013

    • Author(s)
      H. Komori, T. Kajikawa, K. Kataoka, Y. Higuchi, and T. Sakurai
    • Journal Title

      Biochem. Biophys. Res. Commun

      Volume: 438 Pages: 686-690

    • DOI

      10.1016/j.bbrc.2013.07.121

    • Peer Reviewed
  • [Journal Article] Role of the Hydrogen Bond Connecting the Ligands for Substrate and Type I Copper in the Cuprous Oxidase2013

    • Author(s)
      K. Kataoka and T. Sakurai
    • Journal Title

      CueO, Chem. Lett

      Volume: 42 Pages: 1102-1104

    • DOI

      10.1246/cl.130422

    • Peer Reviewed
  • [Journal Article] Modifications of Laccase Activities of Copper Efflux Oxidase, CueO by Synergistic Mutations in the First and Second Coordination Spheres of the Type I Copper Center2013

    • Author(s)
      K. Kataoka, H. Kogi, S. Tsujimura, and T. Sakurai
    • Journal Title

      Biochem. Biophys. Res. Commun

      Volume: 431 Pages: 393-397

    • DOI

      10.1016/j.bbrc.2013.01.040

    • Peer Reviewed
  • [Journal Article] Modifications on the Hydrogen Bond Network by Mutations of Escherichia coli Copper Efflux Oxidase Affect the Process of Proton Transfer to Dioxygen Leading to Alterations of Enzymatic2012

    • Author(s)
      T. Kajikawa, K. Kataoka and T. Sakurai
    • Journal Title

      Activities, Biochem. Biophys. Res. Commun

      Volume: 422 Pages: 152-156

    • DOI

      10.1016/j.bbrc.2012.04.127

    • Peer Reviewed
  • [Journal Article] An O-Centered Structure of the Trinuclear Copper Center in the Cys500Ser/Glu506Gln Mutant of CueO and Structural Changes in Low to High X-Ray Dose Conditions2012

    • Author(s)
      H. Komori, R. Sugiyama, K. Kataoka, Y. Higuchi, and T. Sakurai
    • Journal Title

      Angewandte Chemie

      Volume: 51 Pages: 1861-1864

    • DOI

      10.1002/anie.201107739

    • Peer Reviewed
  • [Journal Article] Enhancement of Laccase Activity through the Construction and Breakdown of a Hydrogen Bond at the Type I Copper Center in Escherichia coli CueO and the Deletion Mutant△α5-7 CueO2011

    • Author(s)
      K. Kataoka, S. Hirota, Y. Maeda, H. Kogi, N. Shinohara, M. Sekimoto, and T. Sakurai
    • Journal Title

      Biochemistry

      Volume: 50 Pages: 558-565

    • DOI

      10.1021/bi101107c

    • Peer Reviewed
  • [Presentation] マルチ銅オキシダーゼの酸素4電子還元における第4の電子ドナーの探索2013

    • Author(s)
      押川直美
    • Organizer
      錯体化学会第63回討論会
    • Place of Presentation
      琉球大学
    • Year and Date
      20131102-04
  • [Presentation] Clostridium属細菌由来の新規マルチ銅オキシダーゼの異種発現と性質2013

    • Author(s)
      原唯歩
    • Organizer
      錯体化学会第63回討論会
    • Place of Presentation
      琉球大学
    • Year and Date
      20131102-04
  • [Presentation] ビリルビンオキシダーゼの酸素還元部位近傍に配置されたTrp132への変異導入2013

    • Author(s)
      押川直美
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      20130911-13
  • [Presentation] マルチ銅酸化酵素における酸素還元反応中心への水素結合ネットワークによるプロトン輸送2013

    • Author(s)
      片岡邦重
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      20130911-13
  • [Presentation] マルチ銅酸化酵素CueOにおける酸素還元反応中心へのプロトン輸送経路2013

    • Author(s)
      片岡邦重
    • Organizer
      2013年度酵素補酵素研究会
    • Place of Presentation
      立命館大学
    • Year and Date
      20130621-22
  • [Presentation] Contribution from a tryptophan residue stacked on the histidine ligand to a type III copper in multicopper oxidases on the four-electron reduction process of dioxygen2013

    • Author(s)
      山本萌
    • Organizer
      第23回金属の関与する生体関連反応シンポジウム
    • Place of Presentation
      武蔵野大学
    • Year and Date
      20130621-22
  • [Presentation] マルチ銅オキシダーゼCueOの三核銅部位近傍に配置された酸性アミノ酸残基の酸素4電子還元における役割2013

    • Author(s)
      櫻井武
    • Organizer
      第40回生体分子科学討論会
    • Place of Presentation
      大阪大学
    • Year and Date
      20130607-08
  • [Presentation] Clostridium属細菌に由来する新規金属オキシダーゼの異種発現と性質2013

    • Author(s)
      原唯歩
    • Organizer
      日本農芸化学会2013年度大会
    • Place of Presentation
      東北大学
    • Year and Date
      20130324-27
  • [Presentation] Clostridium属細菌ゲノムに見出された新規2ドメインラッカーゼの異種発現2012

    • Author(s)
      原唯歩
    • Organizer
      第85回日本生化学会大会
    • Place of Presentation
      マリンメッセ博多
    • Year and Date
      20121214-16
  • [Presentation] Modifications of Hydrogen Bond Network by Mutations of CueO Affect the Process of Proton Transfer to Dioxygen Leading to Alterations of Enzymatic Activities2012

    • Author(s)
      櫻井武
    • Organizer
      錯体化学会第62回討論会
    • Place of Presentation
      富山大学
    • Year and Date
      20120921-23
  • [Presentation] マルチ銅酸化酵素CueOにおける酸素還元反応中心へのプロトン輸送2012

    • Author(s)
      片岡邦重
    • Organizer
      2012年度酵素補酵素研究会
    • Place of Presentation
      名古屋大学
    • Year and Date
      20120727-28
  • [Presentation] Enhancement in Enzymatic Activities for the Copper Efflux Oxidase, CueO by Tuning the Redox Potential of Type I Cu2012

    • Author(s)
      片岡邦重
    • Organizer
      The 12th Japan-China-Korea Joint Symposium on Enzyme Engineering
    • Place of Presentation
      金沢市
    • Year and Date
      20120528-31
  • [Presentation] Type I銅部位の電位制御によるマルチ銅オキシダーゼCueOの反応特性改変2011

    • Author(s)
      小木裕貴
    • Organizer
      第84回日本生化学会大会
    • Place of Presentation
      京都国際会議場
    • Year and Date
      20110921-24
  • [Presentation] 好熱性細菌Geobacillus kaustophilus由来マルチ銅オキシダーゼの異種発現と諸性質2011

    • Author(s)
      中佐英利加
    • Organizer
      第84回日本生化学会大会
    • Place of Presentation
      京都国際会議場
    • Year and Date
      20110921-24
  • [Presentation] マルチ銅オキシダーゼCueOのタイプI銅第二配位圏への変異導入による酸化還元電位の制御と酵素活性2011

    • Author(s)
      小木裕貴
    • Organizer
      錯体化学会第61回討論会
    • Place of Presentation
      岡山理科大学
    • Year and Date
      20110917-19
  • [Book] New Jersey, Multicopper Proteins, in Copper-Oxygen Chemistry, (Eds by K.D. Karlin and S. Itoh)2011

    • Author(s)
      T. Sakurai and K. Kataoka
    • Total Pages
      131-168
    • Publisher
      John Wiley &Sons, Inc.

URL: 

Published: 2015-07-16  

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