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Meta-enzymatic search for D-amino acid oxidoreductases for a novel D-amino acid analysis method.

Research Project

Project/Area Number 15K18684
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Applied biochemistry
Research InstitutionAkita Prefectural University

Principal Investigator

Mutaguchi Yuta  秋田県立大学, 生物資源科学部, 助教 (30724314)

Project Period (FY) 2015-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
KeywordsD-アミノ酸 / 好熱菌 / 酵素分析 / 酸化還元酵素 / 脱水素酵素 / 酸化酵素 / 有用酵素 / メタエンザイム
Outline of Final Research Achievements

The purpose of this work was to find a novel D-amino acid oxidoreductase with the high substrate specificity and stability. We tried to cultivate several thermophile that grow under different nutrient source and pH conditions form different hot spring soils collected in Akita and around. As a result, 10 types of thermophilic flora and 84 thermophilic strains were obtained. Using the activity staining method, we screened D-amino acid oxidoreductase activities from these thermophilic flora and strains. However, no activity detected. In addition, characterization of putative D-amino acid dehydrogenase gene (Locus tag: GK1399) found in genome database of Geobacillus kaustophilus was conducted. The recombinant protein expressed from this gene showed D-amino acid oxidoreductase activities for 7 types of D-amino acid. But, their activities were much lower than other D-amino acid oxidoreductase ready known. Therefore, this enzyme was not a novel D-amino acid oxidoreductase that we aimed at.

Academic Significance and Societal Importance of the Research Achievements

本研究では秋田県及びその周辺の4つ温泉の12カ所の土壌から、様々な栄養源・pHで生育する10種の菌叢と84株の好熱菌株を得ることに成功した。これまで秋田県の温泉に生息する好熱菌を生物資源として活用した研究例は殆ど無かった。よって、本研究において秋田県の温泉から様々な培養条件で生育する好熱菌を見出したことは、秋田の温泉の生物資源としての有用性を示す成果である。今後、有用酵素や生理活性物質の探索に秋田県及びその周辺の温泉から得られた好熱菌が活用されることが期待できる。

Report

(4 results)
  • 2018 Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (8 results)

All 2017 2016 2015

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

  • [Journal Article] Purification of a protein exhibiting isoleucine 2-epimerase activity from Lactobacillus otakiensis JCM 150402015

    • Author(s)
      Yuta Mutaguchi and Toshihisa Ohshima
    • Journal Title

      Bio-protocol

      Volume: 5

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Geobacillus kaustophilus D-アミノ酸脱水素酵素遺伝子の機能解析2015

    • Author(s)
      菅澤達希、牟田口祐太
    • Journal Title

      秋田県立大学ウェブジャーナルB

      Volume: 2 Pages: 62-66

    • Related Report
      2015 Research-status Report
    • Open Access
  • [Presentation] 発酵食品における乳酸菌のD-アミノ酸生産への関与と乳酸菌のD-分岐鎖アミノ酸生産2017

    • Author(s)
      牟田口祐太, 大森勇門, 大島敏久, 春日和, 小嶋郁夫
    • Organizer
      秋田応用生命科学研究会第30回講演会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] D-Amino acid productions and a novel amino acid racemase in lactic acid bacteria2017

    • Author(s)
      Yuta Mutaguchi
    • Organizer
      The Eighth International Conference on Materials Engineering for Resources 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 新規アミノ酸ラセマーゼ, イソロイシン 2-エピメラーゼの構造解析2016

    • Author(s)
      林順司, 牟田口祐太, 大森勇門, 大島敏久, 櫻庭春彦
    • Organizer
      第68回日本ビタミン学会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2016-06-17
    • Related Report
      2016 Research-status Report
  • [Presentation] 乳酸菌Lactobacillus otakiensisにおけるD-分岐鎖アミノ酸の局在と機能2016

    • Author(s)
      十時繁幸, 前田純平, 牟田口祐太, 大島敏久, 土居克実
    • Organizer
      日本農芸化学会2016年度大会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2016-03-28
    • Related Report
      2015 Research-status Report
  • [Presentation] Isoleucine 2-epimeraseホモログ遺伝子をもつ乳酸菌のD-アミノ酸生産2016

    • Author(s)
      牟田口祐太, 春日和, 小嶋郁夫
    • Organizer
      日本農芸化学会2016年度大会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2016-03-28
    • Related Report
      2015 Research-status Report
  • [Book] D-Amino Acids2016

    • Author(s)
      Yuta Mutaguchi, Jyunpei Kobayashi, Tadao Oikawa, and Toshihisa Ohshima
    • Total Pages
      15
    • Publisher
      Springer
    • Related Report
      2016 Research-status Report

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Published: 2015-04-16   Modified: 2020-03-30  

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