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Elucidating the molecular mechanisms of adaptive evolution from bacterial nutrient acquisition strategies

Research Project

Project/Area Number 17H03794
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied microbiology
Research InstitutionNara Institute of Science and Technology

Principal Investigator

Yoshida Shosuke  奈良先端科学技術大学院大学, 研究推進機構, 特任准教授 (80610766)

Project Period (FY) 2017-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2020: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2019: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2018: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2017: ¥7,930,000 (Direct Cost: ¥6,100,000、Indirect Cost: ¥1,830,000)
KeywordsPETase / Ideonella sakaiensis / 遺伝子破壊 / 酵素 / 代謝 / トランスクリプトーム / プロテオーム / 進化 / 細菌
Outline of Final Research Achievements

Ideonella sakaiensis is capable of growing on PET as a carbon nutrition. In this study, we explored the mechanism of efficient PET degradation by I. sakaiensis and found things as follows. (1) Mutations were introduced into the substrate binding site of PETase, and it was found that the high PET substrate specificity of PETase is greatly influenced by these amino acid residues. (2) We succeeded in constructing a gene disruption system for I. sakaiensis, and created Δpetase and Δmhetase strains. (3) Proteomic analysis revealed an enzyme with hydrolytic activity against PET oligomers.

Academic Significance and Societal Importance of the Research Achievements

(1)PETaseのPETに対する基質特異性の高さに貢献するアミノ酸残基が明らかとなった。これらを他のPET加水分解酵素に置換導入することで、機能改良につながることが期待できる。(2)I. sakaiensis遺伝子破壊系により、本菌の持つ遺伝子の生理学的な機能解析を行うことが可能となった。また、本菌の代謝系を人工的に改良し、有用物質生産につなげるなど、代謝工学への応用が可能となった。(3)新酵素の発見により、本菌のPET代謝への理解を進めることができるようになった。

Report

(5 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (26 results)

All 2021 2020 2019 2018 2017

All Journal Article (6 results) (of which Peer Reviewed: 4 results) Presentation (16 results) (of which Int'l Joint Research: 3 results,  Invited: 13 results) Book (3 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Ideonella sakaiensis, PETase, and MHETase: From identification of microbial PET degradation to enzyme characterization2021

    • Author(s)
      Yoshida Shosuke、Hiraga Kazumi、Taniguchi Ikuo、Oda Kohei
    • Journal Title

      Methods in Enzymology

      Volume: 648 Pages: 187-205

    • DOI

      10.1016/bs.mie.2020.12.007

    • ISBN
      9780128220122
    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Microbial Poly(Ethylene Terephthalate) Degradation2020

    • Author(s)
      小田耕平、吉田昭介
    • Journal Title

      Sen'i Gakkaishi

      Volume: 76 Issue: 3 Pages: P-113-P-118

    • DOI

      10.2115/fiber.76.P-113

    • NAID

      130007808450

    • ISSN
      0037-9875, 1884-2259
    • Year and Date
      2020-03-10
    • Related Report
      2020 Annual Research Report
  • [Journal Article] Biodegradation of waste PET2020

    • Author(s)
      Hiraga Kazumi, Taniguchi Ikuo, Yoshida Shosuke, Kimura Yoshiharu, Oda Kohei
    • Journal Title

      EMBO reports

      Volume: 21 Issue: 2

    • DOI

      10.15252/embr.201949826

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Preparation of engineered extracellular vesicles with full-length functional PD-1 membrane proteins by baculovirus expression system2020

    • Author(s)
      Ishikawa Raga、Yoshida Shosuke、Sawada Shin-ichi、Sasaki Yoshihiro、Akiyoshi Kazunari
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: - Issue: 4 Pages: 967-972

    • DOI

      10.1016/j.bbrc.2020.03.187

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Biodegradation of PET: Current Status and Application Aspects2019

    • Author(s)
      Taniguchi Ikuo、Yoshida Shosuke、Hiraga Kazumi、Miyamoto Kenji、Kimura Yoshiharu、Oda Kohei
    • Journal Title

      ACS Catalysis

      Volume: 9 Issue: 5 Pages: 4089-4105

    • DOI

      10.1021/acscatal.8b05171

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] ポリエチレンテレフタレート分解菌・分解酵素の発見とその応用展望2017

    • Author(s)
      吉田昭介、小田耕平
    • Journal Title

      科学と工業

      Volume: 91 Pages: 113-119

    • NAID

      40021210538

    • Related Report
      2017 Annual Research Report
  • [Presentation] Bioprospection of plastic degrading bacteria for the sustainable environment2020

    • Author(s)
      Shosuke Yoshida
    • Organizer
      International Forum on Life Sciences and Education (IFoLSE)
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Poly(ethylene terephthalate)分解菌の代謝メカニズムとその利用2020

    • Author(s)
      吉田昭介
    • Organizer
      第84回日本生化学会中部支部例会・シンポジウム
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Ideonella sakaiensisにおけるPcaG遺伝子破壊株の性状解析2020

    • Author(s)
      西井太郎、蜂須賀真一、吉田昭介
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] PET 分解細菌Ideonella sakaiensisにおけるaldehyde dehydrogenase遺伝子の機能解明2020

    • Author(s)
      蜂須賀真一、高山暁生、藤原毅、吉田昭介
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] ポリエチレンテレフタレート代謝細菌のメカニズムの解明と利用2019

    • Author(s)
      吉田昭介
    • Organizer
      第20回 酵素応用シンポジウム
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] プラスチック分解細菌のエネルギー獲得戦略2019

    • Author(s)
      吉田昭介
    • Organizer
      第71回日本生物工学会大会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 細菌によるPET代謝と、その有用化合物生産への利用展望2019

    • Author(s)
      吉田昭介
    • Organizer
      第68回高分子討論会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] ポリエチレンテレフタレート資化菌の代謝機構とその利用展望2019

    • Author(s)
      吉田昭介
    • Organizer
      19-2エコマテリアル研究会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Bacterial poly(ethylene terephthalate) metabolism by Ideonella sakaiensis2019

    • Author(s)
      Shosuke Yoshida
    • Organizer
      ASP-19 Fifth International Symposium on Advances in Sustainable Polymers
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Ideonella sakaiensisのPETを炭素源とした培養における網羅的遺伝子発現解析2019

    • Author(s)
      高山暁生、吉田昭介
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] ポリエチレンテレフタレート分解細菌の発見、およびその代謝機構の解明2019

    • Author(s)
      吉田昭介
    • Organizer
      「化学コミュニケーションのフロンティア」第4回若手シンポジウム
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] PET資化細菌の発見とメカニズムの解明、及びその応用への展開2019

    • Author(s)
      吉田昭介
    • Organizer
      酵素工学研究会第81回講演会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] PET分解菌~その分解の仕組みと利用展望と・・~2018

    • Author(s)
      吉田昭介
    • Organizer
      第1回創発マテリアル研究会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] A bacterium that metabolizes poly(ethylene terephthalate)2018

    • Author(s)
      Shosuke Yoshida
    • Organizer
      The 10th symposium on International Collaborative Laboratories BioTechnology Institute, U. Minnesota & Grad School BioScience, NAIST
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] ポリエステル分解菌~その分解の仕組みと利用展望~2018

    • Author(s)
      吉田昭介
    • Organizer
      第6回奈良まほろば産学官連携懇話会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] PET樹脂を代謝する細菌2017

    • Author(s)
      吉田昭介
    • Organizer
      BioJapan 2017
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Book] 細菌の代謝能力によるPETの分解(生分解,バイオマスプラスチックの開発と応用)2020

    • Author(s)
      吉田昭介
    • Total Pages
      560
    • Publisher
      技術情報協会
    • ISBN
      9784861047824
    • Related Report
      2019 Annual Research Report
  • [Book] ポリエチレンテレフタレート(PET)分解酵素の発見と構造解析(生分解性プラスチックの環境配慮設計指針)2019

    • Author(s)
      吉田昭介
    • Total Pages
      291
    • Publisher
      シーエムシー出版
    • ISBN
      9784781314877
    • Related Report
      2019 Annual Research Report
  • [Book] 第7 節 既存プラスチックの生分解(マイクロプラスチック問題等各種環境汚染と規制強化に向けたプラスチックの環境対応技術 ~バイオマスプラスチック・生分解性プラスチック・リサイクル・代替~)2019

    • Author(s)
      吉田昭介
    • Total Pages
      306
    • Publisher
      情報機構
    • ISBN
      9784865021714
    • Related Report
      2019 Annual Research Report
  • [Patent(Industrial Property Rights)] ポリヒドロキシアルカン酸の製造方法及び製造用組成物2020

    • Inventor(s)
      吉田昭介、福居俊昭
    • Industrial Property Rights Holder
      奈良先端科学技術大学院大学、東京工業大学
    • Industrial Property Rights Type
      特許
    • Filing Date
      2020
    • Related Report
      2020 Annual Research Report

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Published: 2017-04-28   Modified: 2023-12-25  

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