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Development of a novel system for expression of termite cellulases and its application

Research Project

Project/Area Number 16K14879
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Applied microbiology
Research InstitutionThe University of Tokyo

Principal Investigator

Arioka Manabu  東京大学, 大学院農学生命科学研究科(農学部), 准教授 (20242159)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywordsシロアリ / 共生微生物 / 糸状菌 / セロビオヒドロラーゼ / キシラナーゼ / マンナナーゼ / グルクロン酸エステラーゼ / シロアリおよび共生微生物 / セルラーゼ / バイオマス / シロアリ共生原生生物
Outline of Final Research Achievements

The expression of EuCBH, a cellobiohydrolase from a termite symbiotic protist, was attempted. When the enzyme was expressed in A. oryzae expressing AaBGL1, a beta-glucosidase (BG) from A. aculeatus, increased production was observed compared to the control strain not expressing AaBGL1. The expression of PaBG1b, a BG from wood-eating cockroach, fungal glucuronoyl esterases (GEs), and mannanases from termite symbiotic protists was also conducted. As results, PaBG1b and an A. fumigatus GE were shown to display higher catalytic efficiency than known enzymes. The importance of Trp residue located near the subsite -5 for the affinity to the substrate was also elucidated. Next, the expression and characterization of NtSymX11, an endoxylanase originated from a symbiotic bacterium in the gut of higher termite, were conducted. It was shown that the variants possessing the catalytic domain display 20-times higher catalytic performance compared to another endoxylanase with similar structure.

Academic Significance and Societal Importance of the Research Achievements

オオシロアリ共生原生生物由来セロビオヒドロラーゼEuCBHについては活性の確認には至らなかったが、本研究で新たに取得したbeta-グルコシダーゼPaBG1b、グルクロン酸エステラーゼAfGE、キシラナーゼNtSymX11については従来の酵素よりも高い比活性を有する酵素であることがわかり、今後期待されるバイオマス分解系構築において有力な候補であることを明らかにすることができた。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (13 results)

All 2019 2018 2017 2016

All Journal Article (5 results) (of which Peer Reviewed: 5 results,  Acknowledgement Compliant: 1 results) Presentation (8 results)

  • [Journal Article] Characterization of CBM36-containing GH11 endoxylanase NtSymX11 from the hindgut metagenome of higher termite Nasutitermes takasagoensis displaying prominent catalytic activity.2019

    • Author(s)
      Kitamoto, M., Tokuda, G., Watanabe, H., and Arioka, M.
    • Journal Title

      Carbohydr. Res.

      Volume: 474 Pages: 1-7

    • DOI

      10.1016/j.carres.2019.01.003

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Trp residue at subsite - 5 plays a critical role in the substrate binding of two protistan GH26 beta-mannanases from a termite hindgut2018

    • Author(s)
      Hsu, Y., Koizumi, H., Otagiri, M., Moriya, S., and Arioka, M.
    • Journal Title

      Appl. Microbiol. Biotechnol.

      Volume: 102 Issue: 4 Pages: 1737-1747

    • DOI

      10.1007/s00253-017-8726-2

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] A novel glucuronoyl esterase from Aspergillus fumigatus-the role of conserved Lys residue in the preference for 4-O-methyl glucuronoyl esters2018

    • Author(s)
      Huynh, H.H., Ishii, N., Matsuo, I., and Arioka, M.
    • Journal Title

      Appl. Microbiol. Biotechnol.

      Volume: 102 Issue: 5 Pages: 2191-2201

    • DOI

      10.1007/s00253-018-8739-5

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Heterologous expression in Pichia pastoris and characterization of a β-glucosidase from the xylophagous cockroach Panesthia angustipennis spadica displaying high specific activity for cellobiose2017

    • Author(s)
      Li,Y., Arakawa, G., Tokuda, G., Watanabe, H. & Arioka, M.
    • Journal Title

      Enzyme and Microbial Technology

      Volume: 97 Pages: 104-113

    • DOI

      10.1016/j.enzmictec.2016.11.004

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] Functional expression and characterization of a glucuronoyl esterase from the fungus Neurospora crassa: identification of novel consensus sequences containing the catalytic triad2016

    • Author(s)
      Huynh, H.H. and Arioka, M.
    • Journal Title

      J. Gen. Appl. Microbiol.

      Volume: 62 Pages: 217-224

    • NAID

      130005284314

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] Heterologous production and analysis of cellular responses to the expression of GH11 xylanase from a symbiotic protist in the lower termite2019

    • Author(s)
      Minh Trang PHAM, Gaku TOKUDA, Hirofumi WATANABE, Minoru YOSHIDA, Manabu ARIOKA
    • Organizer
      日本農芸化学会2019年度大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] A study on the enzymatic properties and physiological functions of mannan-degrading enzymes in Neurospora crassa2019

    • Author(s)
      Yunhan HSU, Minoru YOSHIDA, Manabu ARIOKA
    • Organizer
      日本農芸化学会2019年度大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Analysis on the physiological function of glucuronoyl esterase (GE) in the model filamentous fungus Neurospora crassa2019

    • Author(s)
      Ruijie WANG, Hiep Hung HUYNH, Minoru YOSHIDA, Manabu ARIOKA
    • Organizer
      日本農芸化学会2019年度大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] シロアリおよび糸状菌由来の植物バイオマス分解酵素の性質解明とそれらを利用したバイオマス分解系構築の試み2018

    • Author(s)
      有岡 学
    • Organizer
      (公財)発酵研究所 第12回助成研究報告会
    • Related Report
      2018 Annual Research Report
  • [Presentation] シロアリ腸内共生細菌由来GH11キシラナーゼの生産と機能解析2018

    • Author(s)
      北本真理奈、徳田岳、渡辺裕文、有岡学
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Research-status Report
  • [Presentation] Heterologous expression and characterization of GH26 beta-mannanases from a symbiotic protist community of the termite Reticulitermes speratus2017

    • Author(s)
      Yunhan HSU, Hirohiko Koizumi, Masato Otagiri, Shigeharu Moriya, Manabu Arioka
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都女子大学(京都府京都市)
    • Year and Date
      2017-03-17
    • Related Report
      2016 Research-status Report
  • [Presentation] Functional expression and characterization of fungal glucuronoyl esterases2017

    • Author(s)
      HiepHung Huynh, Manabu Arioka
    • Organizer
      日本農芸化学会2017年度大会
    • Place of Presentation
      京都女子大学(京都府京都市)
    • Year and Date
      2017-03-17
    • Related Report
      2016 Research-status Report
  • [Presentation] Characterization of a glucuronoyl esterase (GE) from Aspergillus fumigatus: the role of Lys209 in the preference of 4-O-methyl group in the substrate2017

    • Author(s)
      Hung Hiep Huynh, Nozomi Ishii, Ichiro Matsuo, and Manabu Arioka
    • Organizer
      第17回糸状菌分子生物学コンファレンス
    • Related Report
      2017 Research-status Report

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Published: 2016-04-21   Modified: 2020-03-30  

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