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Development and application of expression system for active selenoproteins

Research Project

Project/Area Number 18K19189
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 38:Agricultural chemistry and related fields
Research InstitutionRitsumeikan University

Principal Investigator

Mihara Hisaaki  立命館大学, 生命科学部, 教授 (30324693)

Co-Investigator(Kenkyū-buntansha) 戸部 隆太  立命館大学, 生命科学部, 講師 (00758823)
Project Period (FY) 2018-06-29 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Fiscal Year 2019: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywordsセレンタンパク質 / タンパク質生産系 / SECIS / セレノシステイン
Outline of Final Research Achievements

Selenoproteins are expected to be useful in industries. Selenoproteins contains a selenocysteine residue in their active sites. Because there are several differences in the mechanisms of selenoprotein biosynthesis among Eukarya, Bacteria, and Archaea, microbial overproduction of heterologous selenoproteins has not been established. In this study, we constructed a luciferase-based reporter assay system that is useful for exploring SECISs that function in Escherichia coli. We examined the function of 9 SECIS structure from 9 selenoproteins genes from an obligately anaerobic bacterium using this reporter assay system in E. coli. We showed that 5 SECIS among 9 can be used as functional SECISs in E. coli.

Academic Significance and Societal Importance of the Research Achievements

セレンを有するタンパク質(セレンタンパク質)には抗酸化活性や糖尿病に関わるものなどがあり、医学・薬学的にも重要である。セレンタンパク質は哺乳類から細菌、アーキアに至るまで様々な生物に多種類見いだされている。セレンタンパク質を大量に生産することが可能となれば、産業利用が加速されると期待されるが、これまで効率の良いセレンタンパク質生産系は確立されていない。本研究では、産業用タンパク質の生産に広く使われている大腸菌における様々なセレンタンパク質の生産系の基盤を構築する成果を得た。

Report

(3 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • Research Products

    (32 results)

All 2020 2019 2018 Other

All Int'l Joint Research (2 results) Journal Article (3 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 3 results,  Open Access: 1 results) Presentation (25 results) (of which Int'l Joint Research: 6 results,  Invited: 4 results) Remarks (2 results)

  • [Int'l Joint Research] Thapar Institute of Eng. Technol.(インド)

    • Related Report
      2019 Annual Research Report
  • [Int'l Joint Research] Tapar Institute of Eng. Technol.(インド)

    • Related Report
      2018 Research-status Report
  • [Journal Article] Selenite uptake by outer membrane porin ExtI and its involvement in the subcellular localization of rhodanese-like lipoprotein ExtH in Geobacter sulfurreducens2019

    • Author(s)
      Jahan MI, Juengwiwattanakitti P, Izu Y, Tobe R, Imai T, Mihara H
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 516 Issue: 2 Pages: 474-479

    • DOI

      10.1016/j.bbrc.2019.06.037

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Apolipoprotein E-mediated regulation of selenoprotein P transportation via exosomes2019

    • Author(s)
      Jin, Y., Chung, Y. W., Jung, M. K., Lee, J. H., Ko, K. Y., Jang, J. K., Ham, M., Kang, H., Pack, C. G., Mihara, H., and Kim, I. Y.
    • Journal Title

      Cell Mol Life Sci

      Volume: - Issue: 12 Pages: 2367-2386

    • DOI

      10.1007/s00018-019-03287-y

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Delivery of selenium to selenophosphate synthetase for selenoprotein biosynthesis2018

    • Author(s)
      Tobe Ryuta、Mihara Hisaaki
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - General Subjects

      Volume: 1862 Issue: 11 Pages: 2433-2440

    • DOI

      10.1016/j.bbagen.2018.05.023

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] 転写調節因子 NsrR のモリブデン補因子生合成における役割2020

    • Author(s)
      藤田大樹, 西田亮, 戸部隆太 三原久明
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] セレン耐性菌 Cellulomonas sp. D3a 株における亜セレン酸還元機構の解析2020

    • Author(s)
      戸部隆太, 牧村康平, 坂本暁紀, T. N. Prakash, 三原久明
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Pseudomonas putida KT2440株由来D-リジントランスポーターによる基質認識2020

    • Author(s)
      三原久明, S. Champas, 山内美咲, 戸部隆太
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 大腸菌におけるセレン酸・テルル酸還元関連遺伝子群の解析2019

    • Author(s)
      藤田大樹, 森聡美, 西田亮, 安間友香理, 田島寛隆, 戸部隆太, 三原久明
    • Organizer
      第36回日本微量栄養素学会学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 大腸菌におけるセレン酸・テルル酸還元関連遺伝子群の解析2019

    • Author(s)
      藤田大樹, 戸部隆太, 三原久明
    • Organizer
      メタルバイオサイエンス研究会2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] Bacillus sp. NTP-1由来新奇DMSOレダクターゼファミリー酵素の研究2019

    • Author(s)
      葛野侑香, 戸部隆太, 三原久明
    • Organizer
      酵素・補酵素研究会2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] Geobacter sulfurreducens由来のロダネーゼ様酵素ExtHの精製条件検討と諸性質解析2019

    • Author(s)
      石戸雄大, 山根祥延, 戸部隆太, 三原久明
    • Organizer
      第62回日本生化学会近畿支部例会
    • Related Report
      2019 Annual Research Report
  • [Presentation] ペルオキシダーゼ活性を有するTruncated hemoglobin (YjbI) の過酸化水素に対する反応2019

    • Author(s)
      田中麻衣, 今井岳志, 戸部隆太, 三原久明
    • Organizer
      日本生物高分子学会2019年度大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Bacillus属細菌のテルル酸還元酵素の同定2019

    • Author(s)
      渡邊樹, 生田帆河, 戸部隆太, 川本純, 栗原達夫, 広瀬侑, 今井友也, N. T. Prakash, 三原久明
    • Organizer
      第30回日本微量元素学会学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Bacillus subtilisのテルル酸還元メカニズムの解明2019

    • Author(s)
      渡邊樹, 戸部隆太, 三原久明
    • Organizer
      メタルバイオサイエンス研究会2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] タンパク質の工学的創製と高活性セレンタンパク質2019

    • Author(s)
      戸部隆太, 川上百香, 三原久明
    • Organizer
      メタルバイオサイエンス研究会2019
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Geobacter sulfurreducensのマルチヘムセレンタンパク質型亜セレン酸還元酵素の機能・構造解析2019

    • Author(s)
      戸部隆太, 吉澤拓也, 伊豆由記子, 吉村文太, 松村浩由, 三原久明
    • Organizer
      第5回日本セレン研究会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 大腸菌ジヒドロピリミジンデヒドロゲナーゼが触媒する複数反応2019

    • Author(s)
      大場杏奈, 吉田吉孝, 石田哲夫, 戸部隆太, 三原久明
    • Organizer
      日本ビタミン学会第71回大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 大腸菌ジヒドロピリミジンデヒドロゲナーゼの触媒特性について2019

    • Author(s)
      三原久明, 大場杏奈, 吉田吉孝, 石田哲夫, 戸部隆太
    • Organizer
      第457回ビタミンB研究協議会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Pseudomonas putidaのD-リジントランスポーターの基質特異性2019

    • Author(s)
      三原久明
    • Organizer
      第458回ビタミンB研究協議会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Function and structure of multiheme-containing selenoprotein2019

    • Author(s)
      Ryuta Tobe, T. Yoshizawa, Y. Izu, M. I. Jahan, B. Yoshimura, H. Matsumura, H. Mihara
    • Organizer
      The 8th International Selenium Conference (Se2019)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Role of outer membrane porin ExtI in selenite reduction in a metal-reducing bacterium2019

    • Author(s)
      H. Mihara, M. I. Jahan, P. Juengwiwattanakitti, Y. Izu, R. Tobe
    • Organizer
      The 8th International Selenium Conference (Se2019)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] A novel selenite-reducing system in the dissimilatory metal-reducing bacterium Geobacter sulfurreducens2019

    • Author(s)
      H. Mihara, M. I. Jahan, M. Jinno, N. Shimamoto, Y. Ishido, P. Juengwiwattanakitti, Y. Izu, R. Tobe
    • Organizer
      Goldschmidt 2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Selenium metabolism in bacteria2019

    • Author(s)
      H. Mihara
    • Organizer
      The 1st International Workshop on Metallomoics and Nanoparticles
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Interaction between rhodanese-related sulfurtransferase ExtH and outer membrane porin ExtI in Geobacter sulfurreducens2019

    • Author(s)
      P. Juengwiwattanakitti, M. I. Jahan, Y. Izu, Y. Ishido, R. Tobe, H. Mihara
    • Organizer
      第5回日本セレン研究会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Purification of glyoxylate oxidase from Gluconobacter dioxyacetonicus A152019

    • Author(s)
      K. Jakkaphan, R. Tobe, H. Mihara
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] The NsrR is essential for the function of selenate/tellurate reductase via activation of the moeA gene in Escherichia coli2019

    • Author(s)
      Daiki FUJITA, Ryo NISHIDA, Ryuta TOBE, H. MIHARA
    • Organizer
      The 1st International Workshop on Metallomoics and Nanoparticles
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Substrate specificity of DlcA of D-lysine ABC transporter from Psuedomonas putida2019

    • Author(s)
      S. Champas, M. Yamauchi, R. Tobe, H. Mihara
    • Organizer
      日本農芸化学会関西・中部支部2019年度合同神戸大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Characterization of the substrate-binding protein DlcA of D-lysine transporter from Pseudomonas putida KT24402019

    • Author(s)
      S. Champas, M. Yamauchi, R. Tobe, H. Mihara
    • Organizer
      第36回日本微量栄養素学会学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Functional analysis of thioredoxin in selenium delivery in bacteria2018

    • Author(s)
      Tobe, R., Shimizu, A., Hagita, S., Ogawa, T., Hirose, Y., Prakash, N. T., Kurihara, T., and Mihara, H.
    • Organizer
      The 7th International Selenium Conference (Se2018)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Remarks] 立命館大学 研究者学術情報 データベース

    • URL

      http://research-db.ritsumei.ac.jp/Profiles/64/0006361/profile.html

    • Related Report
      2019 Annual Research Report
  • [Remarks] 研究者データベース

    • URL

      http://research-db.ritsumei.ac.jp/Profiles/64/0006361/profile.html

    • Related Report
      2018 Research-status Report

URL: 

Published: 2018-07-25   Modified: 2021-02-19  

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