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Modification of the protein secretory system in wood rot fungi

Research Project

Project/Area Number 18H02254
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 40020:Wood science-related
Research InstitutionKyoto University

Principal Investigator

Honda Yoichi  京都大学, 農学研究科, 教授 (70252517)

Co-Investigator(Kenkyū-buntansha) 中沢 威人  京都大学, 農学研究科, 助教 (80608141)
河内 護之  京都大学, 農学研究科, 特定助教 (70771294)
Project Period (FY) 2018-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000)
Fiscal Year 2021: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2020: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2019: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2018: ¥10,270,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥2,370,000)
Keywords木材腐朽菌 / リグニン分解 / タンパク質分泌系 / 担子菌 / 分泌系 / 細胞内小器官 / 白色腐朽菌 / タンパク質分泌 / 木材分解酵素 / 遺伝子組換え / リグニン分解酵素 / 分泌酵素 / ゲノム編集 / 糖鎖修飾
Outline of Final Research Achievements

A gap has been reported repeatedly between the transcription level and actual enzyme activity in the culture filtrate during wood degradation by wood-rot fungi. And activation of protein secretion pathway is proposed to be important for acceleration of lignin degradation by these fungi. In this project, we tried to determine the potential bottle necks in the secretion system and to remove them using genome editing technique to be developed. It was suggested a post-transcriptional regulation may exist that hampers overexpression of recombinant lignin modifying enzyme genes, but there is no evidence of this kind of regulation. We made various chimeric fluorecent proteins combined with potential signal peptides from proteins that may exist in organela of protein secretion pathway. And successfully observed these organera using marking dyes. Moreover, we have developed the first genome editing system in wood rot fungi, introducing CRISPR/Cas9 system in Pleurotus ostreatus.

Academic Significance and Societal Importance of the Research Achievements

持続的社会に必要な再生産可能なバイオマス資源のうちでも、木質資源は食料とも競合しない未来型の資源として注目されている。木質資源を糖化処理して様々な化成品に変換することができれば、材料や燃料として用いる他に、脱化石資源を推進できる多用途な利用が可能である。その際、糖化の妨げとなるのが、難分解性の化合物であるリグニンの存在である。このリグニンを常温常圧でエコフレンドリーに分解できるのが白色腐朽菌と呼ばれる一群のきのこの仲間である。白色腐朽菌のリグニン分解システムの強化において、組換えを用いて木材分解系を強化する事が必要であり、本研究では、そのボトルネックの解明と解決を目指して研究が行われた。

Report

(5 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (15 results)

All 2022 2021 2020 2019 2018

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (13 results) (of which Int'l Joint Research: 10 results,  Invited: 6 results)

  • [Journal Article] Gene targeting using pre-assembled Cas9 ribonucleoprotein and split-marker recombination in <i>Pleurotus ostreatus</i>2021

    • Author(s)
      Boontawon Tatpong、Nakazawa Takehito、Xu Haibo、Kawauchi Moriyuki、Sakamoto Masahiro、Honda Yoichi
    • Journal Title

      FEMS Microbiology Letters

      Volume: 368 Issue: 13 Pages: 1-1

    • DOI

      10.1093/femsle/fnab080

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Efficient genome editing with CRISPR/Cas9 in Pleurotus ostreatus2021

    • Author(s)
      Boontawon Tatpong、Nakazawa Takehito、Inoue Chikako、Osakabe Keishi、Kawauchi Moriyuki、Sakamoto Masahiro、Honda Yoichi
    • Journal Title

      AMB Express

      Volume: 11 Issue: 1 Pages: 1-1

    • DOI

      10.1186/s13568-021-01193-w

    • NAID

      120006960305

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] Genome editing and functional analysis of wood-rot fungus, Pleurotus ostreatus2022

    • Author(s)
      Yoichi Honda, Iori Yamaguchi, Fuga Yamasaki, Takehito Nakazawa, Moriyuki Kawauchi, Masahiro Sakamoto, Minji Oh
    • Organizer
      ICBE2021
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Genome editing in mushroom-forming fungi with CRISPR/Cas9 technology2022

    • Author(s)
      Yoichi Honda;Fuga Yamasaki;Daishiro Koshi;Takeshi Kobukata;Takehito Nakazawa;Moriyuki Kawauchi;Masahiro Sakamoto;Minji Oh
    • Organizer
      KSM 50th anniversary international meeting
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Genome editing to fight with, not against, post-genomics in mushroom fungi2020

    • Author(s)
      Yoichi Honda
    • Organizer
      IUMS2020
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] CRISPR/Cas9 in mushrooms without integration of ectopic DNA2020

    • Author(s)
      Chikako INOUE, Dong Xuan NGUYEN, Takehito NAKAZAWA, & Masahiro SAKAMOTO Yoichi HONDA
    • Organizer
      ECFG2020
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Distribution and shape of organelles visualized with fluorescent proteins in the agaricomycete Pleurotus ostreatus2019

    • Author(s)
      Shivani, Takehito Nakazawa, Masahiro Sakamoto, Yoichi Honda
    • Organizer
      Asia Mycological Congress
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Distribution and shape of organelles visualized with fluorescent proteins in the agaricomyete Pleurotus ostreatus2019

    • Author(s)
      Shivani, Takehito Nakazawa, Masahiro Sakamoto, Yoichi Honda
    • Organizer
      シンポジウム IFOが繋ぐ京大微生物学のフロントライン
    • Related Report
      2019 Annual Research Report
  • [Presentation] Molecular genetics and genome editing in white rot fungus, Pleurotus ostreatus2018

    • Author(s)
      Yoichi Honda
    • Organizer
      Xiamen Forum on Biomass Frontiers
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Molecular genetic approaches beyond the post-genomic era2018

    • Author(s)
      Yoichi Honda
    • Organizer
      9th International Conference on Mushroom Biology and Mushroom Products
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Genome editing with CRISPR/Cas9 system in Pleurotus ostreatus: toward non-GM molecular breeding2018

    • Author(s)
      Takehito Nakazawa
    • Organizer
      SPIRITS 2018 Workshop on ‘Genetic diversity and molecular breeding in cultivated mushrooms’
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] ヒラタケ及びCeriporiopsis subvermisporaにおけるCRISPR/Cas9によるゲノム編集系の確立2018

    • Author(s)
      井上 智香子,中沢 威人, Dong Xuan NGUYEN,刑部 敬史,坂本 正弘, 本田 与一
    • Organizer
      日本きのこ学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] ヒラタケにおける分泌型レポーターアッセイシステムの構築2018

    • Author(s)
      坂 知奈美,中沢 威人,湯村 直樹,竹中 敦紀,Dong Xuan Nguyen, 坂本 正弘,本田 与一
    • Organizer
      日本きのこ学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Efficient Gene Disruption with CRISPR/Cas9 system in Pleurotus ostreatus and Ceriporiopsis subvermispora2018

    • Author(s)
      Chikako INOUE, Takehito NAKAZAWA, Xuan Dong NGUYEN, Keishi OSAKABE, Masahiro SAKAMOTO, and Yoichi HONDA
    • Organizer
      9th International Conference on Mushroom Biology and Mushroom Products
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Reporter Assay System for Secretory Pathway in Pleurotus ostreatus2018

    • Author(s)
      Chinami SAKA, Takehito NAKAZAWA, Naoki YUMURA, Atsuki TAKENAKA, Dong Xuan NGUYEN, Masahiro SAKAMOTO and Yoichi HONDA
    • Organizer
      9th International Conference on Mushroom Biology and Mushroom Products
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research

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Published: 2018-04-23   Modified: 2023-01-30  

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