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Development and applications of yeast-based biosensor for high efficient production of valuable products

Research Project

Project/Area Number 17K17878
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Biofunction/Bioprocess
Applied molecular and cellular biology
Research InstitutionKobe University

Principal Investigator

Nakamura Yasuyuki  神戸大学, 科学技術イノベーション研究科, 特命助教 (40733184)

Project Period (FY) 2017-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsバイオセンサ / 出芽酵母 / シグナル伝達 / G蛋白質共役型受容体 / メラトニン / レポーター遺伝子 / 発酵 / Gタンパク質共役型受容体 / 酵母 / バイオテクノロジー / 微生物
Outline of Final Research Achievements

The aim of this study is to develop the biosensor for the rapid identification of high-producing microbes from large genetic libraries. We tried to evaluate the response of the biosensor for the productivity of the constructed strains. In this study, we created the melatonin-producing yeast strain in which the melatonin biosynthetic pathway was artificially reconstructed, and the constructed metabolite sensor was applied to evaluate the productivity of melatonin. As a result, this system displayed fluorescence in response to the produced melatonin and the intensities were varied depending on the productivity. Combining metabolic engineering with evolutionary engineering, the melatonin sensor constructed will be possible to apply for the rapid breeding of melatonin high-producing yeast strains.

Academic Significance and Societal Importance of the Research Achievements

本研究では、目的物質であるメラトニンのバイオセンサを構築し、メラトニン生合成代謝経路を導入することで、メラトニンの生産量に応じて酵母細胞がセンシングを行うシステムの開発に成功した。本研究で構築したメラトニンセンサを用いることで、メラトニン高生産能力を示す菌株育種への応用が期待される。本研究で用いたメタボライトセンサ構築技術は、メラトニン以外の有用化合物のセンサ構築にも応用できるものであり、様々な高付加価値物質の高生産株の育種技術開発のためのフレームワークを提供する。

Report

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

    (25 results)

All 2018 2017 Other

All Journal Article (6 results) (of which Peer Reviewed: 5 results,  Open Access: 1 results) Presentation (18 results) (of which Int'l Joint Research: 7 results,  Invited: 2 results) Remarks (1 results)

  • [Journal Article] Deletion of DNA ligase IV homolog confers higher gene targeting efficiency on homologous recombination in Komagataella phaffii2018

    • Author(s)
      Ito Yoichiro、Watanabe Toru、Aikawa Shimpei、Nishi Teruyuki、Nishiyama Tozo、Nakamura Yasuyuki、Hasunuma Tomohisa、Okubo Yuji、Ishii Jun、Kondo Akihiko
    • Journal Title

      FEMS Yeast Research

      Volume: 18 Issue: 7

    • DOI

      10.1093/femsyr/foy074

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A Stable, Autonomously Replicating Plasmid Vector Containing Pichia pastoris Centromeric DNA2018

    • Author(s)
      Nakamura Yasuyuki、Nishi Teruyuki、Noguchi Risa、Ito Yoichiro、Watanabe Toru、Nishiyama Tozo、Aikawa Shimpei、Hasunuma Tomohisa、Ishii Jun、Okubo Yuji、Kondo Akihiko
    • Journal Title

      Applied and Environmental Microbiology

      Volume: 84 Issue: 15

    • DOI

      10.1128/aem.02882-17

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Modifying Expression Modes of Human Neurotensin Receptor Type 1 Alters Sensing Capabilities for Agonists in Yeast Signaling Biosensor2018

    • Author(s)
      Hashi Hiroki、Nakamura Yasuyuki、Ishii Jun、Kondo Akihiko
    • Journal Title

      Biotechnology Journal

      Volume: 13 Issue: 4

    • DOI

      10.1002/biot.201700522

    • Related Report
      2018 Annual Research Report 2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Biosensing techniques in yeast: G-protein signaling and protein-protein interaction assays for monitoring ligand stimulation and oligomer formation of heterologous GPCRs2018

    • Author(s)
      Nakamura Yasuyuki、Kondo Akihiko、Ishii Jun
    • Journal Title

      IntechOpen, Peripheral Membrane Proteins

      Volume: - Pages: 25-48

    • Related Report
      2018 Annual Research Report 2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Positive Feedback Genetic Circuit Incorporating a Constitutively Active Mutant Gal3 into Yeast GAL Induction System2017

    • Author(s)
      Ryo Shintaro、Ishii Jun、Matsuno Toshihide、Nakamura Yasuyuki、Matsubara Daiki、Tominaga Masahiro、Kondo Akihiko
    • Journal Title

      ACS Synthetic Biology

      Volume: 6 Issue: 6 Pages: 928-935

    • DOI

      10.1021/acssynbio.6b00262

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] 低分子抗体医薬の開発展望2017

    • Author(s)
      中村泰之、伊藤洋一郎、梅津光央、石井純、近藤昭彦
    • Journal Title

      バイオインダストリー

      Volume: 34 Pages: 54-62

    • Related Report
      2017 Research-status Report
  • [Presentation] Yeast-based in vivo metabolite sensor using signal transduction machinery2018

    • Author(s)
      Jun Ishii, Takuya Tabata, Yasuyuki Nakamura, Akihiko Kondo
    • Organizer
      The 1st international symposium of the Asian Synthetic Biology Association 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Accumulation of novel effective factors enhances the secretion of proteins from Pichia pastoris2018

    • Author(s)
      Akihiko Kondo, Yoichiro Ito, Yasuyuki Nakamura, Shimpei Aikawa, Tomohisa Hasunuma, Jun Ishii
    • Organizer
      i-BioN 2018 The Symposium on Biorefinery and Bioprocess Topics
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Construction of a stable, autonomously replicating plasmid vector containing Pichia pastoris centromeric DNA2018

    • Author(s)
      Yasuyuki Nakamura, Teruyuki Nishi, Risa Noguchi, Yoichiro Ito, Toru Watanabe, Tozo Nishiyama, Shimpei Aikawa, Tomohisa Hasunuma, Jun Ishii, Yoshihiko Yasohara, Akihiko Kondo
    • Organizer
      i-BioN 2018 The Symposium on Biorefinery and Bioprocess Topics
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 酵母細胞における生理活性物質メラトニンをモニタリングするためのGタンパク質共役型受容体(GPCR)を用いたメタボライトセンサの開発2018

    • Author(s)
      田畑 琢也, 中村 泰之, 加藤 寛子, 近藤 昭彦, 石井 純
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] P. pastorisにおける自律複製型ベクターを用いた効率的なDNAマルチアセンブル法2018

    • Author(s)
      西 輝之, 西山 陶三, 山路 大樹, 玉井 雅也, 渡邉 徹, 中村 泰之, 伊藤 洋一郎, 石井 純, 近藤 昭彦, 八十原 良彦
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 遺伝子組換え効率向上に向けたDNAリガーゼIV欠損Pichia pastoris株の開発2018

    • Author(s)
      伊藤 洋一郎, 渡邉 徹, 藍川 晋平, 西 輝之, 西山 陶三, 中村 泰之, 蓮沼 誠久, 八十原 良彦, 石井 純, 近藤 昭彦
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Pichia pastorisにおけるセントロメアDNA配列を用いた自律複製型プラスミドベクターの開発2018

    • Author(s)
      中村 泰之, 西 輝之, 野口 理紗, 伊藤 洋一郎, 渡邉 徹, 西山 陶三, 藍川 晋平, 蓮沼 誠久, 石井 純, 八十原 良彦, 近藤 昭彦
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 発酵制御に向けたグルコースを感知するin vivoバイオセンサーの構築2018

    • Author(s)
      橋弘樹、中村泰之、近藤昭彦、石井純
    • Organizer
      第3回デザイン生命工学研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] A new autonomous replicating plasmid vector containing Pichia pastoris centromeric DNA2018

    • Author(s)
      Nakamura Yasuyuki、Nishi Teruyuki、Noguchi Risa、Ito Yoichiro、Watanabe Toru、Nishiyama Tozo、Aikawa Shimpei、Hasunuma Tomohisa、Ishii Jun、Yasohara Yoshihiko、Kondo Akihiko
    • Organizer
      The 9th International Symposium of Innovative BioProduction Kobe
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Development of G-protein-coupled receptor (GPCR)-based metabolite sensor for monitoring physiological chemical melatonin in eukaryotic yeast cells2018

    • Author(s)
      Tabata Takuya、Nakamura Yasuyuki、Kondo Akihiko、Ishii Jun
    • Organizer
      The 9th International Symposium of Innovative BioProduction Kobe
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Enhanced protein secretion by accumulation of novel effective factors in Pichia pastoris2018

    • Author(s)
      Ito Yoichiro、Nakamura Yasuyuki、Nishi Teruyuki、Aikawa Shimpei、Hasunuma Tomohisa、Ishii Jun、Kondo Akihiko
    • Organizer
      The 9th International Symposium of Innovative BioProduction Kobe
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] A simple and efficient DNA multi-assemble method using episomal vector in Pichia pastoris2018

    • Author(s)
      Nishi Teruyuki、Tamai Masaya、Yamaji Taiki、Watanabe Toru、Nishiyama Tozo、Nakamura Yasuyuki、Ito Yoichiro、Ishii Jun、Kondo Akihiko、Yasohara Yoshihiko
    • Organizer
      The 9th International Symposium of Innovative BioProduction Kobe
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] P. pastorisにおける自律複製型ベクターを用いた効率的なDNAマルチアセンブル法2018

    • Author(s)
      西輝之、山路大樹、玉井雅也、渡邉徹、西山陶三、中村泰之、伊藤洋一郎、石井純、近藤昭彦、八十原良彦
    • Organizer
      第12回日本ゲノム微生物学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 特定の生理活性物質を濃度依存的に感知する酵母メタボライトセンサの開発2017

    • Author(s)
      田畑琢也、中村泰之、石井純、近藤昭彦
    • Organizer
      生物工学若手研究者の集い 夏のセミナー2017
    • Related Report
      2017 Research-status Report
  • [Presentation] Pichia pastorisにおけるセントロメア配列を含む自律複製型プラスミドの開発2017

    • Author(s)
      中村泰之、西輝之、野口理紗、伊藤洋一郎、渡邉徹、西山陶三、藍川晋平、蓮沼誠久、石井純、八十原良彦、近藤昭彦
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] Pichia pastorisのタンパク質分泌生産における有用因子の獲得とその蓄積による効果の検証2017

    • Author(s)
      伊藤洋一郎、中村泰之、西輝之、藍川晋平、蓮沼誠久、石井純、近藤昭彦
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] メラトニン濃度をモニタリングするための酵母in vivoメタボライトセンサ2017

    • Author(s)
      田畑琢也、中村泰之、石井純、近藤昭彦
    • Organizer
      化学工学会 第49回秋季大会
    • Related Report
      2017 Research-status Report
  • [Presentation] ヒト由来生理活性物質を特異的に検知する酵母in vivoバイオセンサ2017

    • Author(s)
      田畑琢也、中村泰之、石井純、近藤昭彦
    • Organizer
      神戸大学 研究基盤センター 若手フロンティア研究会
    • Related Report
      2017 Research-status Report
  • [Remarks] 神戸大学大学院科学技術イノベーション研究科バイオ生産工学研究室ホームページ

    • URL

      http://www2.kobe-u.ac.jp/~akondo/index.html

    • Related Report
      2018 Annual Research Report 2017 Research-status Report

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Published: 2017-04-28   Modified: 2020-03-30  

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