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Generation of the cell factories for bio-based organic acid production through improvement of filamentous fungi by metabolic engineering

Research Project

Project/Area Number 22580093
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied microbiology
Research InstitutionWaseda University

Principal Investigator

KIRIMURA Kohtaro  早稲田大学, 理工学術院, 教授 (90195412)

Project Period (FY) 2010 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2010: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Keywords応用微生物 / 酵素 / セルファクトリ / 代謝工学 / 有機酸 / クエン酸 / シュウ酸 / トランスアコニット酸 / 発酵 / バイオテクノロジー / クロコウジカビ / ポリケタイド
Research Abstract

For generation of cell factories for bio-based organic acid production, generation of strains showing high gene-targeting frequencies was performed in citric acid-producing Aspergillus niger. Novel type III polyketide synthase and some key enzymes, such as methylcitrate synthase and aconitate isomerase, in relation to the tricarboxylic acid cycle were examined through gene cloning and enzymatic characterization. There is some possibility that heterologous expression of genes encoding novel and specific enzymes in the citric acid-producing strains as hosts and improvement by metabolic engineering will open the way for generation of the cell factories for bio-based organic acid production.

Report

(4 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (47 results)

All 2013 2012 2011 2010

All Journal Article (12 results) (of which Peer Reviewed: 12 results) Presentation (31 results) Book (4 results)

  • [Journal Article] p-Aminosalicylic Acid Production by Enzymatic KolbeSchmitt Reaction Using Salicylic Acid Decarboxylases Improved through Site-Directed Mutagenesis.2013

    • Author(s)
      Ienaga, S., Kosaka, S., Honda, Y., Ishii, Y., Kirimura, K.
    • Journal Title

      Bull. Chem. Soc. Jpn.

      Volume: 86 Pages: 628-634

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Gene Identification and Functional Analysis of Methylcitrate Synthase in Citric Acid-Producing Aspergillus niger WU-2223L Biosci2013

    • Author(s)
      Kobayashi, K., Hattori, T., Honda, Y., Kirimura, K.
    • Journal Title

      Biotech. Biochem.

      Volume: (In press)

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] p-Aminosalicylic Acid Production by Enzymatic Kolbe-Schmitt Reaction Using Salicylic Acid Decarboxylases Improved through Site-Directed Mutagenesis2013

    • Author(s)
      Saori Ienaga
    • Journal Title

      Bulletin of the Chemical Society of Japan

      Volume: In press Issue: 5 Pages: 628-634

    • DOI

      10.1246/bcsj.20130006

    • NAID

      10031167857

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Visual Expression Analysis of the Responses of the Alternative Oxidase Gene (aox1) to Heat Shock, Oxidative, and Osmotic Stresses in Conidia of Citric Acid-Producing Aspergillus niger.2012

    • Author(s)
      Honda, Y., Hattori, T., Kirimura, K.
    • Journal Title

      J. Biosci. Bioeng.

      Volume: 113 Pages: 338-342

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Visual Expression Analysis of the Responses of the Alternative Oxidase Gene (aox1) to Heat Shock, Oxidative, and Osmotic Stresses in Conidia of Citric Acid-Producing A.niger2012

    • Author(s)
      Yuki Honda, Takasumi Hattori, Kohtaro Kirimura
    • Journal Title

      J.Biosci.Bioeng.

      Volume: 113 Issue: 3 Pages: 338-342

    • DOI

      10.1016/j.jbiosc.2011.10.026

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Increases in Gene-Targeting Frequencies Due to Disruption of kueA as a ku80 Homolog in Citric Acid-Producing Aspergillus niger.2011

    • Author(s)
      Honda, Y.,Kobayashi, K., Kirimura, K.
    • Journal Title

      Biosci. Biotech. Biochem.

      Volume: 75 Pages: 1594-1596

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] High-yield Production of cis,cis-Muconic Acid from Catechol in Aqueous Solution by Biocatalyst.2011

    • Author(s)
      Kaneko, A., Ishii, Y., Kirimura, K.
    • Journal Title

      Chem. Lett.

      Volume: 40 Pages: 381-383

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Production of p-Aminosalicylic Acid through Enzymatic Kolbe-Schmitt Reaction Catalyzed by Reversible Salicylic Acid Decarboxylase.2011

    • Author(s)
      Kirimura, K., Yanaso, S., Kosaka, S., Koyama, K., Hattori, T., Ishii, I.
    • Journal Title

      Chem. Lett.

      Volume: 40 Pages: 206-208

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] High-yield Production of cis, cis-Muconic Acid from Catechol in Aqueous Solution by Biocatalyst2011

    • Author(s)
      Aya Kaneko, Yoshitaka Ishii, Kohtaro Kirimura
    • Journal Title

      Chem.Lett.

      Volume: 40 Issue: 4 Pages: 381-383

    • DOI

      10.1246/cl.2011.381

    • NAID

      10028170617

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Increases in Gene-Targeting Frequencies Due to Disruption of kueA as a ku80 Homolog in Citric Acid-Producing Aspergillus niger2011

    • Author(s)
      Yuki Honda, Keiichi Kobayashi, Kohtaro Kirimura
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 75 Issue: 8 Pages: 1594-1596

    • DOI

      10.1271/bbb.110015

    • NAID

      10029591872

    • ISSN
      0916-8451, 1347-6947
    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Production of p-Aminosalicylic Acid through Enzymatic Kolbe-Schmitt Reaction Catalyzed by Reversible Salicylic Acid Decarboxylase2011

    • Author(s)
      K.Kirimura, S.Yanaso, S.Kosaka, K.Koyama, T.Hattori, Y.Ishii
    • Journal Title

      Chem.Lett.

      Volume: 40 Pages: 206-208

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Increase of Gene-targeting Frequencies by Disruption of kueA as a ku80 Homolog in Citric Acid-producing Aspergillus niger2011

    • Author(s)
      Y.Honda, K.Kobayashi, K.Kirimura
    • Journal Title

      Biosci.Biotechnol.Biochem.

      Volume: 75(in press)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] アコニット酸イソメラーゼ遺伝子を高発現させた大腸菌によるtrans-アコニット酸生産2013

    • Author(s)
      油原かほり、濱地達也、小林慶一、本田裕樹、桐村光太郎
    • Organizer
      日本農芸化学会2013年度大会
    • Place of Presentation
      仙台
    • Year and Date
      2013-03-27
    • Related Report
      2012 Final Research Report
  • [Presentation] クエン酸の特異的検出を可能とするインジケータ蛍光タンパク質の創製2013

    • Author(s)
      本田裕樹、桐村光太郎
    • Organizer
      日本農芸化学会2013年度大会
    • Place of Presentation
      仙台
    • Year and Date
      2013-03-26
    • Related Report
      2012 Final Research Report
  • [Presentation] 部位特異的変異の導入による可逆的サリチル酸脱炭酸酵素の改変とp- アミノサリチル酸の高収率生産への応用2013

    • Author(s)
      家永里織
    • Organizer
      日本化学会第93春季年回(2013)
    • Place of Presentation
      滋賀
    • Related Report
      2012 Annual Research Report
  • [Presentation] Pseudomonas sp. WU-0701 由来アコニット酸イソメラーゼをコードする遺伝子の大腸菌における異種発現2013

    • Author(s)
      油原かほり
    • Organizer
      日本化学会第93春季年回(2013)
    • Place of Presentation
      滋賀
    • Related Report
      2012 Annual Research Report
  • [Presentation] アコニット酸イソメラーゼ遺伝子を高発現させた大腸菌によるtrans-アコニット酸生産2013

    • Author(s)
      油原かほり
    • Organizer
      日本農芸化学会2013年度大会
    • Place of Presentation
      仙台
    • Related Report
      2012 Annual Research Report
  • [Presentation] 部位特異的変異の導入による可逆的サリチル酸脱炭酸酵素の改変とp-アミノサリチル酸の生産への応用2012

    • Author(s)
      家永里織、伊藤優斗、本田裕樹、石井義孝、桐村光太郎
    • Organizer
      第64回日本生物工学会大会
    • Place of Presentation
      神戸
    • Year and Date
      2012-10-25
    • Related Report
      2012 Final Research Report
  • [Presentation] クエン酸生産糸状菌Aspergillus nigerにおけるメチルクエン酸シンターゼの検出と機能解析2012

    • Author(s)
      小林慶一、本田裕樹、桐村光太郎
    • Organizer
      第64回日本生物工学会大会
    • Place of Presentation
      神戸
    • Year and Date
      2012-10-24
    • Related Report
      2012 Final Research Report
  • [Presentation] Pseudomonas sp. WU-0701由来アコニット酸イソメラーゼをコードする遺伝子の大腸菌における異種発現2012

    • Author(s)
      油原かほり、米原広海、小林慶一、本田裕樹、服部貴澄、桐村光太郎
    • Organizer
      第64回日本生物工学会大会
    • Place of Presentation
      神戸
    • Year and Date
      2012-10-24
    • Related Report
      2012 Final Research Report
  • [Presentation] 低分子ゲル化剤としての機能を示すメントール配糖体の酵素的合成2012

    • Author(s)
      井手浩平、梅澤覚、小林慶一、本田裕樹、桐村光太郎
    • Organizer
      日本化学会第92回春季年会
    • Place of Presentation
      慶應義塾大学(神奈川)
    • Year and Date
      2012-03-27
    • Related Report
      2011 Annual Research Report
  • [Presentation] 糸状菌Aspergillus niger NRRL328由来III型ポリケタイド合成酵素の機能解析2012

    • Author(s)
      濱地達也、小林慶一、本田裕樹、桐村光太郎
    • Organizer
      日本化学会第92回春季年会
    • Place of Presentation
      慶應義塾大学(神奈川)
    • Year and Date
      2012-03-27
    • Related Report
      2011 Annual Research Report
  • [Presentation] 糸状菌Aspergillus nigerにおけるoxaloacetate hydrolase遺伝子の高発現による高収率シュウ酸生産2012

    • Author(s)
      小林慶一、本田裕樹、服部貴澄、桐村光太郎
    • Organizer
      日本農芸化学会2012年度大会
    • Place of Presentation
      京都女子大学(京都)
    • Year and Date
      2012-03-24
    • Related Report
      2011 Annual Research Report
  • [Presentation] Pseudomonas sp.WU-0701由来アコニット酸イソメラーゼの酵素的諸性質の検討と遺伝子クローニング2012

    • Author(s)
      米原広海、本田裕樹、服部貴澄、桐村光太郎
    • Organizer
      日本農芸化学会2012年度大会
    • Place of Presentation
      京都女子大学(京都)
    • Year and Date
      2012-03-24
    • Related Report
      2011 Annual Research Report
  • [Presentation] 組換え大腸菌の細胞反応によるカテコールからのcis,cis -ムコン酸の水系高収率生産2012

    • Author(s)
      宇田川恵右
    • Organizer
      第6回バイオ関連化学シンポジウム
    • Place of Presentation
      札幌
    • Related Report
      2012 Annual Research Report
  • [Presentation] 複数の部位特異的変異を導入した可逆的サリチル酸脱炭酸酵素の改変によるp-アミノサリチル酸の生産2012

    • Author(s)
      家永里織
    • Organizer
      第6回バイオ関連化学シンポジウム
    • Place of Presentation
      札幌
    • Related Report
      2012 Annual Research Report
  • [Presentation] Pseudomonas sp. WU-0701 由来アコニット酸イソメラーゼをコードする遺伝子の大腸菌における異種発現2012

    • Author(s)
      油原かほり
    • Organizer
      第64回日本生物工学会大会
    • Place of Presentation
      神戸
    • Related Report
      2012 Annual Research Report
  • [Presentation] 部位特異的変異の導入による可逆的サリチル酸脱炭酸酵素の改変とp- アミノサリチル酸の生産への応用2012

    • Author(s)
      家永里織
    • Organizer
      第64回日本生物工学会大会
    • Place of Presentation
      神戸
    • Related Report
      2012 Annual Research Report
  • [Presentation] クエン酸生産糸状菌Aspergillus nigerにおけるoxaloacetate hydrolase遺伝子(oahA)の破壊と高発現によるシュウ酸生産経路の検証2011

    • Author(s)
      小林慶一、本田裕樹、服部貴澄、桐村光太郎
    • Organizer
      第11回糸状菌分子生物学コンファレンス
    • Place of Presentation
      東京大学(東京)
    • Year and Date
      2011-11-17
    • Related Report
      2011 Annual Research Report
  • [Presentation] Aspergillus niger NRRL 328由来III型ポリケタイド合成酵素によるピロン化合物の生成2011

    • Author(s)
      濱地達也、宮井希実、小林慶一、本田裕樹、桐村光太郎
    • Organizer
      第11回糸状菌分子生物学コンファレンス
    • Place of Presentation
      東京大学(東京)
    • Year and Date
      2011-11-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] 組換え大腸菌を生体触媒として用いた逐次添加法によるカテコールからのcis,cis-ムコン酸の高収率生産2011

    • Author(s)
      宇田川恵右、金子亞矢、本田裕樹、石井義孝、桐村光太郎
    • Organizer
      第63回日本生物工学会大会
    • Place of Presentation
      東京農工大学(東京)
    • Year and Date
      2011-09-27
    • Related Report
      2011 Annual Research Report
  • [Presentation] Aspergillus niger NRRL328株由来III型ポリケタイド合成酵素ホモログの機能解析2011

    • Author(s)
      濱地達也、宮井希実、小林慶一、本田裕樹、桐村光太郎
    • Organizer
      第63回日本生物工学会大会
    • Place of Presentation
      東京農工大学(東京)
    • Year and Date
      2011-09-26
    • Related Report
      2011 Annual Research Report
  • [Presentation] クエン酸生産糸状菌Aspergillus nigerにおけるメチルクエン酸シンターゼ遺伝子破壊株の作製2011

    • Author(s)
      小林慶一、本田裕樹、桐村光太郎
    • Organizer
      第63回日本生物工学会大会
    • Place of Presentation
      東京農工大学(東京)
    • Year and Date
      2011-09-26
    • Related Report
      2011 Annual Research Report
  • [Presentation] 糸状菌由来の新規なIII型ポリケタイド合成酵素の遺伝子異種発現と機能解析2011

    • Author(s)
      小林慶一、宮井希実、濱地達也、本田裕樹、桐村光太郎
    • Organizer
      第5回バイオ関連化学シンポジウム
    • Place of Presentation
      つくば国際会議場(茨城)
    • Year and Date
      2011-09-14
    • Related Report
      2011 Annual Research Report
  • [Presentation] 部位特異的変異を利用したサリチル酸脱炭酸酵素の改変とカルボキシル化活性の向上2011

    • Author(s)
      梅澤覚、家永里織、小坂祥代、本田裕樹、桐村光太郎
    • Organizer
      節5回バイオ関連化学シンポジウム
    • Place of Presentation
      つくば国際会議場(茨城)
    • Year and Date
      2011-09-13
    • Related Report
      2011 Annual Research Report
  • [Presentation] Production of p-Aminosalicylic Acid by Regioselective Carboxylation through Enzymatic Kolbe-Schmitt Reaction Using Salicylic Acid Decarboxylase2011

    • Author(s)
      Yuki Honda, Sachiyo Kosaka, Yoshitaka Ishii, Kohtaro Kirimura
    • Organizer
      4th Congress of European Microbiologists, FEMS 2011
    • Place of Presentation
      Geneva (Switzerland)
    • Year and Date
      2011-06-28
    • Related Report
      2011 Annual Research Report
  • [Presentation] Disruption of kueA as a ku80 Homolog in Citric Acid-Producing Aspergillus niger for Improvement of Homologous Recombination Frequencies2011

    • Author(s)
      Keiichi Kobayashi, Yuki Honda, Kohtaro Kirimura
    • Organizer
      4th Congress of European Microbiologists, FEMS 2011
    • Place of Presentation
      Geneva (Switzerland)
    • Year and Date
      2011-06-28
    • Related Report
      2011 Annual Research Report
  • [Presentation] Aspergillus niger由来III型ポリケタイド合成酵素の基質特異性と反応生成物2011

    • Author(s)
      宮井希美、米原広海、本田裕樹、服部貴澄、桐村光太郎
    • Organizer
      日本農芸化学会大会(2011年度)
    • Place of Presentation
      京都
    • Year and Date
      2011-03-27
    • Related Report
      2010 Annual Research Report
  • [Presentation] クエン酸生産糸状菌由来ku80破壊株における相同組換え効率の向上2010

    • Author(s)
      本田裕樹、小林慶一、桐村光太郎
    • Organizer
      第10回糸状菌分子生物学コンファレンス
    • Place of Presentation
      広島
    • Year and Date
      2010-11-19
    • Related Report
      2010 Annual Research Report
  • [Presentation] Aspergillus niger由来III型ポリケタイド合成酵素遺伝子の機能解析2010

    • Author(s)
      宮井希美、本田裕樹、服部貴澄、桐村光太郎
    • Organizer
      第10回糸状菌分子生物学コンファレンス
    • Place of Presentation
      広島
    • Year and Date
      2010-11-19
    • Related Report
      2010 Annual Research Report
  • [Presentation] クエン酸生産糸状菌Aspergillus niger由来ku80破壊株の作製と相同組換え効率の向上2010

    • Author(s)
      小林慶一、本田裕樹、桐村光太郎
    • Organizer
      日本生物工学会大会(2010年度)
    • Place of Presentation
      宮崎
    • Year and Date
      2010-10-29
    • Related Report
      2010 Annual Research Report
  • [Presentation] 酵母Trichosporon moniliiformeにおけるサリチル酸脱炭酸酵素が関与する新規なサリチル酸分解経路2010

    • Author(s)
      本田裕樹、服部貴澄、岩崎勇一郎、石井義孝、桐村光太郎
    • Organizer
      日本生物工学会大会(2010年度)
    • Place of Presentation
      宮崎
    • Year and Date
      2010-10-28
    • Related Report
      2010 Annual Research Report
  • [Presentation] 可逆的サリチル酸脱炭酸酵素における部位特異的変異を利用したサリチル酸合成活性の向上2010

    • Author(s)
      小坂祥代、本田裕樹、服部貴澄、桐村光太郎
    • Organizer
      日本生物工学会大会(2010年度)
    • Place of Presentation
      宮崎
    • Year and Date
      2010-10-28
    • Related Report
      2010 Annual Research Report
  • [Book] Bio-based chemicals, Citric acid. In : Murray MY (ed) Comprehensive Biotechnology, 2nd edn. vol. 3.2011

    • Author(s)
      Kirimura, K., Honda, Y., Hattori, T.
    • Related Report
      2012 Final Research Report
  • [Book] Bio-based chemicals, Gluconic and Itaconic Acids. In : Murray MY (ed) Comprehensive Biotechnology, 2nd edn. vol.2011

    • Author(s)
      Kirimura, K., Honda, Y., Hattori, T.
    • Related Report
      2012 Final Research Report
  • [Book] "Citric Acid", Comprehensive Biotechnology, 2nd edition(Ed.Moo-Young)2011

    • Author(s)
      Kohtaro Kirimura, Yuki Honda, Takasumi Hattori
    • Publisher
      Elsevier (London)
    • Related Report
      2011 Annual Research Report
  • [Book] "Gluconic and Itaconic Acids", Comprehensive Biotechnology, 2nd edition(Ed.Moo-Young)2011

    • Author(s)
      Kohtaro Kirimura, Yuki Honda, Takasumi Hattori
    • Publisher
      Elsevier (London)
    • Related Report
      2011 Annual Research Report

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Published: 2010-08-23   Modified: 2019-07-29  

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