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Understanding of luciferin-luciferase system by the artificial evolution of luciferase

Research Project

Project/Area Number 20611005
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Chemical biology
Research InstitutionNagoya University

Principal Investigator

OBA Yuichi  Nagoya University, 生命農学研究科, 助教 (40332704)

Project Period (FY) 2008 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2010: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2009: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2008: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Keywords人工進化 / ルシフェリン / ルシフェラーゼ / ホタル / ヒカリコメツキ / 脂肪酸CoA合成酵素 / 水素結合ネットワーク / 酵素反応
Research Abstract

Site-directed mutagenesis of AbLL, a fatty acyl-CoA synthetase in non-luminous click beetle Agrypnus binodulus, showed that only one-three amino acid substitutions are sufficient to generate the luciferase activity. The activity was weak but the further amino acid changes gave stronger activity, more than 40 times. We also isolated an isotype luciferase, LcLuc2, from the Japanese firefly Luciola cruciata (Oba et al., 2010), suggesting that fireflies possess at least two luciferase genes and that luciferin-binding sites (7 amino acids) in luciferases are highly conserved. Interestingly, the luminescent color by LcLuc2 is pH insensitive, unlike other firefly luciferases.

Report

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

    (23 results)

All 2011 2010 2009 2008 Other

All Journal Article (8 results) (of which Peer Reviewed: 8 results) Presentation (8 results) Book (5 results) Remarks (2 results)

  • [Journal Article] Identification and characterization of a luciferase isotype in the Japanese firefly, Luciola cruciata, involving in the dim glow of firefly egg.2010

    • Author(s)
      Oba Yuichi, Mori Nanae, Yoshida Mayumi, Inouye Satoshi
    • Journal Title

      Biochemistry 49

      Pages: 10788-10795

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Identification and characterization of a luciferase isotype in the Japanese firefly, Luciola cruciata, involving in the dim glow of firefly eggs.2010

    • Author(s)
      Y.Oba, N.Mori, M.Yoshida, S.Inouye
    • Journal Title

      Biochemistry

      Volume: 49 Pages: 10788-10795

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Functional conversion of fatty acyl-CoA synthetase to firefly luciferase by site-directed mutagenesis : A key substitution responsible for luminescence activity.2009

    • Author(s)
      Oba Yuichi, Iida Koichiro, Inouye Satoshi
    • Journal Title

      FEBS Letters 583

      Pages: 2004-2008

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Functional conversion of fatty acyl-CoA synthetase to firefly luciferase by site-directed mutagenesis : A key substitution responsible for luminescence activity.2009

    • Author(s)
      Y.Oba, K.Iida, S.Inouye
    • Journal Title

      FEBS Letters 583

      Pages: 2004-2008

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Determination of the luciferin contents in luminous and non-luminous beetles.2008

    • Author(s)
      Oba Yuichi, Shintani Takeru, Nakamura Takanori, Ojika Makoto, Inouye Satoshi
    • Journal Title

      Biosci.Biotechnol.Biochem. 72(5)

      Pages: 1384-1387

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Orthologous gene of beetle luciferase in non-luminous click beetle, Agrypnus binodulus (Elateridae), encodes a fatty acyl-CoA synthetase.2008

    • Author(s)
      Oba Yuichi, Iida Koichiro, Ojika Makoto, Inouye Satoshi
    • Journal Title

      Gene 407

      Pages: 169-175

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Orthologous gene of beetle luciferase in non-luminous click beetle, Agrypnus binodulus (Elateridae), encodes fatty acyl-CoA synthetase2008

    • Author(s)
      Y. Oba, K. Iida, M. Ojika, S. Inouye
    • Journal Title

      Gene 407

      Pages: 169-175

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Determination of the luciferin contents in luminous and non-lurai nous beetles.2008

    • Author(s)
      Y. Oba, T. Shintani, T. Nakamura, M. Ojika, S. Inouye
    • Journal Title

      Biosci. Biotechnol. Biochem. 72

      Pages: 1384-1387

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Presentation] 脂肪酸CoA合成酵素からルシフェラーゼへの人工進化.2009

    • Author(s)
      大場裕一、飯田幸一郎、井上敏
    • Organizer
      日本動物学会第80回大会
    • Place of Presentation
      グランシップ静岡.
    • Year and Date
      2009-09-19
    • Related Report
      2010 Final Research Report
  • [Presentation] 脂肪酸CoA合成酵素からルシフェラーゼへの人工進化2009

    • Author(s)
      大場裕一、飯田幸一郎、井上敏
    • Organizer
      日本動物学会第80回大会
    • Place of Presentation
      グランシップ静岡
    • Year and Date
      2009-09-19
    • Related Report
      2009 Annual Research Report
  • [Presentation] ヒカリコメツキルシフェラーゼの酵素特性とその進化.2009

    • Author(s)
      熊崎瑞保、大場裕一
    • Organizer
      日本動物学会第80回大会
    • Place of Presentation
      グランシップ静岡.
    • Year and Date
      2009-09-17
    • Related Report
      2010 Final Research Report
  • [Presentation] ヒカリコメツキルシフェラーゼの酵素特性とその進化2009

    • Author(s)
      熊崎瑞保、大場裕一
    • Organizer
      日本動物学会第80回大会
    • Place of Presentation
      グランシップ静岡
    • Year and Date
      2009-09-17
    • Related Report
      2009 Annual Research Report
  • [Presentation] 非発光性コメツキムシからのルシフェラーゼオーソログ遺伝子のクローニングおよびRNAiを用いた機能解析2008

    • Author(s)
      飯田幸一郎, 大場裕一, 小鹿一, 井上敏
    • Organizer
      日本進化学会
    • Place of Presentation
      東京大学
    • Year and Date
      2008-08-23
    • Related Report
      2008 Annual Research Report
  • [Presentation] Orthologous gene of beetle luciferase in non-luminous click beetle.2008

    • Author(s)
      飯田幸一郎, 大場裕一
    • Organizer
      Evolution 2008
    • Place of Presentation
      University of Minnesota
    • Year and Date
      2008-06-23
    • Related Report
      2010 Final Research Report 2008 Annual Research Report
  • [Presentation] Evolutinary origin of beetle luciferase.2008

    • Author(s)
      大場裕一, 井上敏
    • Organizer
      Evolution 2008
    • Place of Presentation
      University of Minnesota.
    • Year and Date
      2008-06-22
    • Related Report
      2010 Final Research Report
  • [Presentation] Evolutional2008

    • Author(s)
      Y. Oba, S. Inouye
    • Organizer
      Evolution 2008
    • Place of Presentation
      University of Minnesota
    • Year and Date
      2008-06-22
    • Related Report
      2008 Annual Research Report
  • [Book] 地球からのおくりもの.(ホタル.)2011

    • Author(s)
      大場裕一
    • Publisher
      風媒社.
    • Related Report
      2010 Final Research Report
  • [Book] 地球からのおくりもの2011

    • Author(s)
      大場裕一
    • Total Pages
      216
    • Publisher
      風媒社
    • Related Report
      2010 Annual Research Report
  • [Book] In "Bioluminescence in Focus-A Collection of Illuminating Essays".(On the Origin of Beetle luminescence.)2009

    • Author(s)
      大場裕一
    • Publisher
      Research Signpost
    • Related Report
      2010 Final Research Report
  • [Book] Bioluminescence in Focus-A collection of illuminating essays.2009

    • Author(s)
      V.B.Meyer-Rochow, et al.
    • Total Pages
      385
    • Publisher
      Research Signpost
    • Related Report
      2009 Annual Research Report
  • [Book] 昆虫ミメティックス〜昆虫の設計に学ぶ〜2008

    • Author(s)
      大場裕一
    • Publisher
      エヌ・ティー・エス
    • Related Report
      2008 Annual Research Report
  • [Remarks] ホームページ等

    • Related Report
      2010 Final Research Report
  • [Remarks]

    • URL

      http://obayuichi.web.fc2.com/index.html

    • Related Report
      2010 Annual Research Report

URL: 

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

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