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Analysis on molecular mechanism for cold adapted glucokinase from psychrophillic bacteria

Research Project

Project/Area Number 18H02412
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43040:Biophysics-related
Research InstitutionSaga University

Principal Investigator

Horitani Masaki  佐賀大学, 農学部, 准教授 (80532134)

Co-Investigator(Kenkyū-buntansha) 渡邉 啓一  佐賀大学, 農学部, 教授 (40191754)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2020: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2019: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2018: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Keywords酵素 / 構造生物化学 / 生物物理 / 電子スピン共鳴 / 低温適応酵素 / X線結晶構造解析 / タンパク質工学 / 電子スピン共鳴法 / スピンラベル / 熱安定酵素 / 生体分子 / 低温適応 / 構造生物学 / 熱安定性 / 生体関連化学 / 構造解析 / 好冷細菌
Outline of Final Research Achievements

We have cloned glucokinase from psychrophillic bacteria which is isolated from shells lived in Antarctic Ocean, and prepared purification system. We also have made purification system for glucokinase from Escherichia coli as comparison. Interestingly, glucokinase from Antarctic Ocean showed higher activity at a wide range of temperatures and higher thermal stability than that from E. coli. Revealing this characteristics for glucokinase, we have applied X-ray crystallographic and site-directed mutagenesis spin label electron spin resonance studies. Our biochemical and biophysical studies have successfully elucidated the molecular mechanism for glucokinase form Antarctic Ocean.

Academic Significance and Societal Importance of the Research Achievements

酵素はアミノ酸から成る環境負荷の無い化学反応触媒として注目されている。一般的な酵素は人工的には高コストが必要な化学反応を低コストで高効率に触媒できることが知られている。ところが、酵素はそれぞれ指摘温度を持ち、それ以外では低活性または失活してしまう。さらに通常の酵素は熱を加えると容易に変性してしまい、元には戻れない。これが酵素の産業利用が進んでいない大きな理由である。本研究で、幅広い温度で高活性で熱安定性の高い酵素を天然で発見した。さらにこの酵素が持つ仕組みの一端を原子レベルで解き明かすことに成功したので、今後は今回得られた知見を検証し、自然界に優しい素材である酵素を用いた産業へと発展させる。

Report

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

    (32 results)

All 2021 2020 2019 2018 Other

All Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 4 results) Presentation (22 results) (of which Int'l Joint Research: 7 results,  Invited: 5 results) Book (1 results) Remarks (3 results)

  • [Journal Article] Biochemical properties of CumA multicopper oxidase from plant pathogen, <i>Pseudomonas syringae</i>2021

    • Author(s)
      Ishida Konan、Tsukamoto Yuya、Horitani Masaki、Ogawa Tomohisa、Tanaka Yoshikazu
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 85 Issue: 9 Pages: 1995-2002

    • DOI

      10.1093/bbb/zbab123

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The molecular mechanism of phytosphingosine binding to FFAR4/GPR120 differs from that of other fatty acids2021

    • Author(s)
      Nagasawa Tomotaka、Horitani Masaki、Kawaguchi Shin‐ichi、Higashiyama Shigeki、Hama Yoichiro、Mitsutake Susumu
    • Journal Title

      FEBS Open Bio

      Volume: 11 Issue: 11 Pages: 3081-3089

    • DOI

      10.1002/2211-5463.13301

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Timing of NO Binding and Protonation in the Catalytic Reaction of Bacterial Nitric Oxide Reductase as Established by Time-Resolved Spectroscopy.2020

    • Author(s)
      Hanae Takeda, Tatsunari Kimura, Takashi Nomura, Masaki Horitani, Azusa Yokota, Akiko Matsubayashi, Shoko Ishii, Yoshitsugu Shiro, Minoru Kubo and Takehiko Tosha
    • Journal Title

      Bulletin of the Chemical Society of Japan

      Volume: - Issue: 7 Pages: 825-833

    • DOI

      10.1246/bcsj.20200038

    • NAID

      130007879554

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] X-ray Crystallography and Electron Paramagnetic Resonance Spectroscopy Reveal Active Site Rearrangement of Cold-Adapted Inorganic Pyrophosphatase2020

    • Author(s)
      Horitani Masaki、Kusubayashi Kazuki、Oshima Kyoka、Yato Akane、Sugimoto Hiroshi、Watanabe Keiichi
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 1-12

    • DOI

      10.1038/s41598-020-61217-6

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] The [4Fe-4S] cluster of sulfurtransferase TtuA desulfurizes TtuB during tRNA modification in Thermus thermophilus2020

    • Author(s)
      Minghao Chen, Masato Ishizaka, Shun Narai, Masaki Horitani, Naoki Shigi, Min Yao & Yoshikazu Tanaka
    • Journal Title

      Communications Biology

      Volume: 3 Issue: 1 Pages: 168-168

    • DOI

      10.1038/s42003-020-0895-3

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] An ancient type of MnmA protein is an iron?sulfur cluster-dependent sulfurtransferase for tRNA anticodons2019

    • Author(s)
      Shigi Naoki、Horitani Masaki、Miyauchi Kenjyo、Suzuki Tsutomu、Kuroki Misao
    • Journal Title

      RNA

      Volume: 26 Issue: 3 Pages: 240-250

    • DOI

      10.1261/rna.072066.119

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Presentation] 急速凍結XAFS法による低温適応無機ピロフォスファターゼの温度特性機構の解明2021

    • Author(s)
      丸岡 早紀, 馬込 栄輔, 瀬戸山 寛之, 河本 正秀, 堀谷 正樹, 渡邉 啓一
    • Organizer
      第24回XAFS討論会
    • Related Report
      2020 Annual Research Report
  • [Presentation] X-ray Crystallography and Site-Directed Spin-Labeling ESR Reveal Cold Adaptation and High Thermal Stability Mechanisms of Cold-Adapted Glucokinase2021

    • Author(s)
      Akane Yato, Rio Asaka, Hiroshi Sugimoto, Keiichi Watanabe, Masaki Horitani
    • Organizer
      22nd International Society of Magnetic Resonance
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Analysis on Cold Adaptation Mechanism of Metalloenzyme by X-ray Crystallography and EPR Spectroscopy Combined with Rapid Freeze-Quench2021

    • Author(s)
      Masaki Horitani, Yuri Kasu, Hiroshi Sugimoto, Keiichi Watanabe
    • Organizer
      22nd International Society of Magnetic Resonance
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 急速凍結EPR法およびX線結晶構造解析法による好冷細菌由来無機ピロフォスファターゼの低温適応機構の解明2021

    • Author(s)
      堀谷 正樹, 杉本 宏, 渡邉 啓一
    • Organizer
      第47回生体分子討論会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 動的・静的構造解析による南極産好冷細菌由来グルコキナーゼの低温適応・高熱安定性機構の解明2021

    • Author(s)
      矢垰 紅音, 浅香 里緒, 杉本 宏, 渡邉 啓一, 堀谷 正樹
    • Organizer
      第47回生体分子討論会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Development of Advanced THz ESR Spectroscopy for Heme Proteins with Integer Spin System2019

    • Author(s)
      Haruna Ninomiya, Tsubasa Okamoto, Kana Nishikawa, Susumu Okubo, Eiji Omichi, Hiroshi Fujii, Hitoshi Ohta, Masaki Horitani
    • Organizer
      Awaji Island Conference on Electron Spin Science & Technology
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] EPR Spectroscopy Reveals the Active Site Rearrangement of Di-Mn2+ Center by Substrate Binding in Inorganic Pyrophosphatase from Shewanella sp. AS-112019

    • Author(s)
      Masaki Horitani, Hiroshi Sugimoto, Keiichi Watanabe
    • Organizer
      Awaji Island Conference on Electron Spin Science & Technology
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Electron Paramagnetic Resonance Reveals Cold Adaption Mechanism of the Psychrophilic Metalloenzyme2019

    • Author(s)
      Masaki Horitani
    • Organizer
      第19回日本蛋白質科学会年会、第71回日本細胞生物学会大会 合同年次大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 先端テラヘルツESR装置開発によるヘムタンパク質研究の新展開2019

    • Author(s)
      二宮 春菜, 岡本 翔, 西川 佳那, 大久保 晋, 大道 英二, 藤井 浩, 太田 仁, 堀谷 正樹
    • Organizer
      九重分子科学セミナー2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] Cold Adaption Inorganic Pyrophosphatase from Shewanella sp. AS-11 was Investigated the Reaction Mechanism by X-ray Crystallography and Electron Paramagnetic Resonance Spectroscopy2019

    • Author(s)
      Masaki Horitani, Hiroshi Sugimoto, Keiichi Watanabe
    • Organizer
      International Congress on Pure & Applied Chemistry (ICPAC) Yangon 2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] ラジカルSAM酵素におけるラジカル反応機構~自然界はどのようにしてラジカルを安全に利用しているか~2019

    • Author(s)
      堀谷 正樹
    • Organizer
      2019年度第一回水和ナノ構造研究会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 好冷細菌由来無機ピロフォスファターゼの低温適応機構2019

    • Author(s)
      堀谷 正樹, 楠林 和貴, 大嶋 杏佳, 杉本 宏, 渡邉 啓一
    • Organizer
      第58回NMR討論会・SEST2019連合大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 好冷細菌由来無機ピロフォスファターゼの低温適応機構・熱安定性機構2019

    • Author(s)
      堀谷 正樹, 杉本 宏, 渡邊 啓一
    • Organizer
      日本生体エネルギー研究会 第45回討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 好冷細菌由来グルコキナーゼの低温適応・高熱安定性機構の解明2019

    • Author(s)
      矢垰 紅音, 浅香 里緒, 渡邊 啓一, 堀谷 正樹
    • Organizer
      日本生体エネルギー研究会 第45回討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] テラヘルツ ESR 法による金属タンパク質の電子状態研究2019

    • Author(s)
      堀谷 正樹
    • Organizer
      平成30年度神戸大学分子フォトサイエンス研究センター共同利用研究成果報告会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Electron Paramagnetic Resonance Spectroscopy Investigates Di-Manganese(II) Active Site of Inorganic Pyrophosphatase from Shewanella sp. AS-112018

    • Author(s)
      Masaki Horitani
    • Organizer
      19th Malaysian International Chemistry Congress and International Congress on Pure & Applied Chemistry Langkawi 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] EPR Studies Reveal Mn(II)-Mn(II) Distance in the Active Site of Inorganic Pyrophosphatase from Shewanella sp. AS-112018

    • Author(s)
      Masaki Horitani
    • Organizer
      The Third Joint Conference of the Asia-Pacific EPR/ESR Society and The International EPR (ESR) Society
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Structural Analysis on di-Mn ion site for Shewanella Inorganic Pyrophosphatase by EPR2018

    • Author(s)
      堀谷 正樹、榊原 由季、渡邉 啓一
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 電子常磁性共鳴法による低温適応酵素無機ピロフォスファターゼの複核Mnイオンサイトの構造解析2018

    • Author(s)
      堀谷 正樹
    • Organizer
      九重分子科学セミナー2018
    • Related Report
      2018 Annual Research Report
  • [Presentation] 低温適応酵素グルコキナーゼの低温適応機構の解明2018

    • Author(s)
      加藤 侑希、林 芙由子、渡邉 啓一、堀谷 正樹
    • Organizer
      第42回蛋白質と酵素の構造と機能に関する九州シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] Elucidation of the tRNA thiolation mechanism of TtuA involved in Fe-S cluster2018

    • Author(s)
      Masato Ishizaka, Minghao Chen, Syun Narai, Masaki Horitani, Seiko Oka, Yoshikazu Tanaka, Min Yao
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 新規な酸素感受性tRNA硫黄化酵素の機能解析2018

    • Author(s)
      鴫 直樹、姫野 美沙緒、堀谷 正樹
    • Organizer
      極限環境生物学会2018年度 (第19回) 年会
    • Related Report
      2018 Annual Research Report
  • [Book] 生命金属ダイナミクス ~生体内における金属の挙動と制御~2021

    • Author(s)
      城 宜嗣、津本 浩平
    • Total Pages
      564
    • Publisher
      (株)エヌ・ティー・エス
    • ISBN
      9784860437060
    • Related Report
      2020 Annual Research Report
  • [Remarks] 佐賀大学農学部生命機能科学科分子生命科学分野堀谷研究室

    • URL

      http://horitani.ag.saga-u.ac.jp/horitanilab/index.html

    • Related Report
      2020 Annual Research Report
  • [Remarks] 佐賀大学農学部生命機能科学科分子生命化学分野堀谷研究室

    • URL

      http://horitani.ag.saga-u.ac.jp/horitanilab/index.html

    • Related Report
      2019 Annual Research Report
  • [Remarks] 佐賀大学農学部生命機能科学科分子生命科学分野

    • URL

      http://horitani.ag.saga-u.ac.jp/horitanilab/index.html

    • Related Report
      2018 Annual Research Report

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

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