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Conversion of an inverting exo-D-glucosaminidase into a golycosynthase

Research Project

Project/Area Number 22780095
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Applied biochemistry
Research InstitutionIshikawa Prefectural University

Principal Investigator

HONDA Yuji  石川県立大学, 生物資源環境学部, 准教授 (40399382)

Project Period (FY) 2010 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2012: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2011: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2010: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsエキソβ-D-グルコサミニダーゼ / エキソ-β-D-グルコサミニダーゼ / グルコサミニダーゼ / Photobacterium / Vibrio / アノマー反転型酵素
Research Abstract

We elucidated an inverting exo-β-D-glucosaminidase activity of the GH9 protein from Photobacterium profundum SS9. Kinetic analysis of the hydrolytic activities toward cellooligosaccharides and chitooligosaccharides indicated the enzyme is an exo-β-D-glucosaminidase (EC 3.2.1.165) rather than an exo-β-1,4-glucosidase.Based on the modelled three-dimensional structure, we targeted D139, D143, and E555 as candidates of the important amino acid residues in the hydrolytic reaction. D139A, D143A and E555A mutants displayed trace hydrolytic activities. The D139A displayed glycosynthase activity with α-GlcN-F as a substrate.

Report

(4 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (15 results)

All 2013 2012 2011 2010

All Journal Article (4 results) (of which Peer Reviewed: 4 results) Presentation (11 results)

  • [Journal Article] Heterogonous expression and characterization of a plant class IV chitinase from the pitcher of the carnivorous plant Nepenthes alata2012

    • Author(s)
      Kana Ishisaki, Yuji Honda, Hajime Taniguchi, Naoya Hatano, and Tatsuro Hamada
    • Journal Title

      Glycobiology

      Volume: Vol.22 Pages: 345-351

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Heterogonous expression and characterization of a plant class IV chitinase from the pitcher of the carnivorous plant Nepenthes alata2012

    • Author(s)
      Kana Ishisaki, et al
    • Journal Title

      Glycobiology

      Volume: 22 Issue: 3 Pages: 345-351

    • DOI

      10.1093/glycob/cwr142

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Elucidation of exo-β-D-glucosaminidase activity of a family 9 glycoside hydrolase (PBPRA0520) from Photobacterium profundum SS92011

    • Author(s)
      Yuji Honda, Nozomi Shimaya, Kana Ishisaki, Mitsuru Ebihara, and Hajime Taniguchi
    • Journal Title

      Glycobiology

      Volume: Vol.21 Pages: 503-511

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Elucidation of exo-β-D-glucosaminidase activity of a family 9 glycoside hydrolase (PBPRA0520) from Photobacterium profundum SS92011

    • Author(s)
      Yuji Honda, et al.
    • Journal Title

      Glycobiology

      Volume: 21 Pages: 503-511

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] GH ファミリー9 に属するエキソβ-D-グルコサミニダーゼの糖鎖合成酵素化2013

    • Author(s)
      本多裕司
    • Organizer
      日本農芸化学会2013年度大会
    • Place of Presentation
      宮城県
    • Related Report
      2012 Annual Research Report 2012 Final Research Report
  • [Presentation] Photobacterium profundum SS9由来エキソβ-D-グルコサミニダーゼの糖鎖合成酵素化2012

    • Author(s)
      新井祥子(本多裕司)
    • Organizer
      日本応用糖質科学会平成24年度大会
    • Place of Presentation
      東京都
    • Related Report
      2012 Annual Research Report 2012 Final Research Report
  • [Presentation] Reaction mechanism of an inverting exo-β-D-glucosaminidase from Photobacterium profundum SS92012

    • Author(s)
      Yuji Honda
    • Organizer
      the 12th Japan-China-Korea Joint Symposium on Enzyme Engineering
    • Place of Presentation
      the 12th Japan-China-Korea Joint Symposium on Enzyme Engineering
    • Related Report
      2012 Final Research Report
  • [Presentation] Reaction mechanism of an inverting exo-β-D-glucosaminidase from Photobacterium profundum SS92012

    • Author(s)
      Yuji Honda et al.
    • Organizer
      the 12th Japan-China-Korea Joint Symposium on Enzyme Engineering
    • Place of Presentation
      石川県
    • Related Report
      2012 Annual Research Report
  • [Presentation] Photobacteriumprofundum SS9 由来エキソ型 β-D-グルコサミニダーゼの変異型酵素の機能解析2011

    • Author(s)
      新井祥子 (本多裕司)
    • Organizer
      2011 年度日本農芸化学会関西・中部支部合同大会
    • Place of Presentation
      京都
    • Year and Date
      2011-10-02
    • Related Report
      2012 Final Research Report
  • [Presentation] 食虫植物ウツボカズラ消化液キチン質分解酵素の機能解析2011

    • Author(s)
      石崎佳奈(本多裕司)
    • Organizer
      2011年度日本農芸化学会関西・中部支部合同大会
    • Place of Presentation
      京都
    • Year and Date
      2011-10-02
    • Related Report
      2012 Final Research Report 2011 Annual Research Report
  • [Presentation] Photobacterium profundum SS9由来エキソ型β-D-グルコサミニダーゼの変異型酵素の機能解析2011

    • Author(s)
      新井祥子, ら
    • Organizer
      2011年度日本農芸化学会関西・中部支部合同大会
    • Place of Presentation
      京都
    • Year and Date
      2011-10-02
    • Related Report
      2011 Annual Research Report
  • [Presentation] Yuji HONDA2011

    • Author(s)
      Yuji HONDA
    • Organizer
      16th European Carbohydrate Symposium
    • Place of Presentation
      Sorrento, Italy
    • Year and Date
      2011-07-04
    • Related Report
      2012 Final Research Report
  • [Presentation] The GH9 protein from Photobacterium profundum SS9 is an exo-β-D-glucosaminidase (EC 3.2.1.165)2011

    • Author(s)
      Yuji HONDA, et al
    • Organizer
      16th European Carbohydrate Symposium
    • Place of Presentation
      Sorrento, Italy
    • Year and Date
      2011-07-04
    • Related Report
      2011 Annual Research Report
  • [Presentation] Photobacterium profundum SS9 由来 GH9 酵素の機能解析2010

    • Author(s)
      本多裕司
    • Organizer
      日本農芸化学会関西支部大会
    • Place of Presentation
      奈良
    • Year and Date
      2010-10-03
    • Related Report
      2012 Final Research Report
  • [Presentation] Photobacterium profundum SS9由来GH9酵素の機能解析2010

    • Author(s)
      本多裕司
    • Organizer
      日本農芸化学会関西支部大会
    • Place of Presentation
      奈良
    • Year and Date
      2010-10-03
    • Related Report
      2010 Annual Research Report

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Published: 2010-08-23   Modified: 2019-07-29  

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