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Development of Glyco-medicines Using Mutant Enzyme of Microbial Endoglycosidase and Establishment of Its Preparation Method

Research Project

Project/Area Number 20380052
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied microbiology
Research InstitutionKyoto University

Principal Investigator

YAMAMOTO Kenji  Kyoto University, 生物資源環境学部, 客員教授 (70109049)

Co-Investigator(Kenkyū-buntansha) ASHIDA Hisashi  京都大学, 大学院・生命科学研究科, 准教授 (40379087)
Project Period (FY) 2008 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥19,110,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥4,410,000)
Fiscal Year 2010: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2009: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2008: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Keywordsエンドグリコシダーゼ / 糖転移活性 / オキサゾリン化合物 / 部位特異的変異酵素 / 化学-酵素合成 / 糖鎖医薬品 / インフルエンザウィルス / 感染阻害剤 / substrate assisted catalysis / サブスタンスP / PAMP12 / リボヌクレアーゼB / シアロ糖鎖 / インフルエンザウイルス / 変異酵素
Research Abstract

The endo-β-N-acetylglucosaminidase from Mucor hiemalis isolated from soil sample is unique in that it can transfer en bloc the oligosaccharide of glycoproteins onto different acceptors, and thereby it enzymatically generates diverse glycoconjugates. We performed mutational studies to obtain the mutant of the enzyme which had significantly diminished hydrolysis activity but had a high transglycosylation activity. We have found that some mutant could efficiently exhibit transglycosylation activity using synthetic sugar oxazoline. Therefore, we used the mutant enzyme to synthesize an efficient binding inhibitor for influenza virus, which is composed of sialyloligosaccharides on chitosan. This compound showed sufficient inhibitory activity against influenza virus infection.

Report

(4 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report
  • 2008 Annual Research Report
  • Research Products

    (36 results)

All 2010 2009 2008 Other

All Journal Article (11 results) (of which Peer Reviewed: 11 results) Presentation (19 results) Book (4 results) Remarks (2 results)

  • [Journal Article] Efficient Transfer of Sialo-oligosaccharides onto Proteins by Combined Use of a Glycosynthase-like Mutant of Mucor hiemalis Endoglycosidase and Synthetic Sialo-complex-type Sugar Oxazoline.2010

    • Author(s)
      M.Umekawa, T.Higashiyama, Y.Koga, T.Tanaka, M.Noguchi, A.Kobayashi, S.Shoda, W.Huang, L.-X.Wang, H.Ashida, K.Yamamoto
    • Journal Title

      Biochim. Biophys. Acta 1800

      Pages: 1203-1209

    • NAID

      120002647341

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Efficient Glycosynthase Mutant Derived from Mucor hiemalis Endo-β-N-acetylglucosaminidase Capable of Transferring Oligosaccharide from Both Sugar Oxazoline and Natural N-Glycan.2010

    • Author(s)
      M.Umekawa, C.Li, T.Higashiyama, W.Huang, H.Ashida, K.Yamamoto, L.-X.Wang
    • Journal Title

      J. Biol. Chem 285(1)

      Pages: 511-521

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Efficient transfer of sialo-oligosaccharide onto proteins by combined use of a glycosynthase-like mutant of Mucor hiemalis endoglycosidase and synthetic sialo-complex-type sugar oxazoline2010

    • Author(s)
      M.Umekawa, et al.
    • Journal Title

      Biochimica et Biophysica Acta

      Volume: 1800 Pages: 1203-1209

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Efficient Glycosynthase Mutant Derived from Mucor hiemalis Endo-β-Nacetylglucosaminidase Capable of Transferring Oligosaccharide from Both Sugar Oxazoline and N-Glycan2010

    • Author(s)
      M.Umekawa, et al.
    • Journal Title

      The Journal of Biological Chemistry 285

      Pages: 511-521

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Glycosynthases Enable a Highly Efficient Chemoenzymatic Synthesis of N-Glycoproteins Carrying Intact Nature N-Glycans.2009

    • Author(s)
      W.Huang, C.Li, B.Li, M.Umekawa, K.Yamamoto, X.Zhang, L.-X.Wang
    • Journal Title

      J. Am. Chem. Soc 131(6)

      Pages: 2214-2223

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Glycosynthases Enable a Highly Efficient Chemoenzymatic Synthesis of N-Glycoproteins Carrying Intact Nature N-Glycans2009

    • Author(s)
      W.Huang, et al.
    • Journal Title

      Journal of The American Chemical Society 131

      Pages: 2214-2223

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Glycosynthases Enable a Highly Efficient Chemoenzymatic Synthesis of N-Glycoproteins Carrying Intact Natural N-Glyeans2009

    • Author(s)
      W. Huang, et al.
    • Journal Title

      Journal of The American Chemical Society 131

      Pages: 2214-2223

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Design of a Sialylglycopolymer with a Chitosan Backbone Having Efficient Inhibitory Activity against Influenza Virus Infection.2008

    • Author(s)
      M.Umemura, M.Itoh, Y.Makimura, K.Yamazaki, M.Umekawa, A.Masui, Y.Matahira, M.Shibata, H.Ashida, K.Yamamoto
    • Journal Title

      J. Med. Chem 51

      Pages: 4496-4503

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Mutants of Mucor hiemalis Endo-β-N-acetylglucosaminidase Show Enhanced Transglycosylation and Glycosynthase-like Activities.2008

    • Author(s)
      M.Umekawa, W.Huang, B.Li, K.Fujita, H.Ashida, L.-X Wang, K.Yamamoto
    • Journal Title

      J. Biol. Chem 283(8)

      Pages: 4469-4479

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Mutants of Mucor hiemlis Endo-β-N-acetylglucosaminidase Show Enhanced Transglycosylation and Glycosynthase-like Activities2008

    • Author(s)
      M. Umekawa, et al.
    • Journal Title

      The Journal of Biological Chemistry 283

      Pages: 4469-4479

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Design of a Sialylglycopolymer with a Chitosan Backbone Having Efficient Inhibitory Activity agaainst Influenza Virus Infection2008

    • Author(s)
      M. Umemura, et al.
    • Journal Title

      Journal of Medicinal Chemstry 51

      Pages: 4496-4503

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Presentation] Endo-M変異酵素を用いた機能性糖鎖複合体の効率的な合成2010

    • Author(s)
      山本憲二
    • Organizer
      グライコバイオロジクス研究会
    • Place of Presentation
      大阪千里ライフサイエンスセンター
    • Year and Date
      2010-12-11
    • Related Report
      2010 Annual Research Report 2010 Final Research Report
  • [Presentation] グライコシンターゼ様エンドグリコシダーゼ変異酵素を用いたシアロ糖ペプチドの合成2010

    • Author(s)
      東山貴幸, ら
    • Organizer
      日本農芸化学会2010年度大会
    • Place of Presentation
      東京大学
    • Year and Date
      2010-03-28
    • Related Report
      2010 Final Research Report
  • [Presentation] グライコシンターゼ様エンドグリコシダーゼ変異酵素を用いたシアロ糖ペプチドの合成2010

    • Author(s)
      東山貴幸, ら
    • Organizer
      日本農芸化学会2010年度大会
    • Place of Presentation
      東京大学、東京都
    • Year and Date
      2010-03-28
    • Related Report
      2009 Annual Research Report
  • [Presentation] ひとつだけの酵素を用いた効率的な糖鎖の付加とリモデリング2009

    • Author(s)
      山本憲二
    • Organizer
      第7回糖鎖科学コンソーシアムシンポジウム
    • Place of Presentation
      大阪千里ライフサイエンスセンター
    • Year and Date
      2009-12-07
    • Related Report
      2010 Final Research Report
  • [Presentation] ひとつだけの酵素を用いた効率的な糖鎖の付加とリモデリング2009

    • Author(s)
      山本憲二
    • Organizer
      第7回糖鎖科学コンソーシアムシンポジウム
    • Place of Presentation
      千里ライフサイエンスセンター、大阪市
    • Year and Date
      2009-12-07
    • Related Report
      2009 Annual Research Report
  • [Presentation] 糸状菌由来エンドグリコシダーゼの機能改変と糖鎖複合体合成への応用2009

    • Author(s)
      東山貴幸, ら
    • Organizer
      日本生物工学会大会第61回大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-24
    • Related Report
      2010 Final Research Report
  • [Presentation] ノイラミニダーゼ阻害剤によるインフルエンザウイルス捕捉型感染阻害剤の効果促進2009

    • Author(s)
      三谷誠司, ら
    • Organizer
      日本生物工学会大会第61回大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-24
    • Related Report
      2010 Final Research Report
  • [Presentation] 糸状菌由来エンドグリコシダーゼの機能改変と糖鎖複合体合成への応用2009

    • Author(s)
      東山貴幸, ら
    • Organizer
      日本生物工学会大会第61回大会
    • Place of Presentation
      名古屋大学、名古屋市
    • Year and Date
      2009-09-24
    • Related Report
      2009 Annual Research Report
  • [Presentation] ノイラミニダーゼ阻害剤によるインフルエンザウイルス捕捉型感染阻害剤の効果促進2009

    • Author(s)
      三谷誠司, ら
    • Organizer
      日本生物工学会大会第61回大会
    • Place of Presentation
      名古屋大学、名古屋市
    • Year and Date
      2009-09-24
    • Related Report
      2009 Annual Research Report
  • [Presentation] 微生物由来エンドグリコシダーゼの機能改変と機能性糖鎖複合体の合成への応用2009

    • Author(s)
      梅川碧里, ら
    • Organizer
      第29回日本糖質学会
    • Place of Presentation
      高山飛騨生活文化センター
    • Year and Date
      2009-09-11
    • Related Report
      2010 Final Research Report
  • [Presentation] 微生物由来エンドグリコシダーゼの機能改変と機能性糖鎖複合体の合成への応用2009

    • Author(s)
      梅川碧里, ら
    • Organizer
      第29回日本糖質学会
    • Place of Presentation
      高山飛騨生活文化センター、岐阜県
    • Year and Date
      2009-09-11
    • Related Report
      2009 Annual Research Report
  • [Presentation] Improvement of a Novel Glycosynthase-like Mutant of Mucor hiemalis Endoglycosidase for Efficient Syntheses of Glycoconjugates2009

    • Author(s)
      梅川碧里, ら
    • Organizer
      15th European Carbohydrate Symposium
    • Place of Presentation
      ウィーン大学、オーストリア国
    • Year and Date
      2009-07-20
    • Related Report
      2010 Final Research Report
  • [Presentation] Improvement of a Novel Glycosynthase-like Mutant of Mucor hiemalis Endoglycosidase for Efficient Syntheses of Glycoconjugates2009

    • Author(s)
      梅川碧里, et al.
    • Organizer
      15^<th> European Carbohydrate Symposium
    • Place of Presentation
      ウィーン大学、オーストリア国
    • Year and Date
      2009-07-20
    • Related Report
      2009 Annual Research Report
  • [Presentation] シアロ糖鎖を活用したインフルエンザウイルス捕捉型感染阻害剤のデザイン2009

    • Author(s)
      梅村舞子, ら
    • Organizer
      日本農芸化学会関西支部講演会
    • Place of Presentation
      京都
    • Year and Date
      2009-02-07
    • Related Report
      2010 Final Research Report 2008 Annual Research Report
  • [Presentation] 糸状菌由来endo-β-N-acetylglucosaminidaseが有する特異な糖転移機能の解析とグライコシンターゼ化2009

    • Author(s)
      梅川碧里, ら
    • Organizer
      日本農芸化学会関西支部講演会
    • Place of Presentation
      京都
    • Year and Date
      2009-02-07
    • Related Report
      2010 Final Research Report 2008 Annual Research Report
  • [Presentation] Development of a Novel Glycosynthase Mutant of Endo-β-N-acetylglucosaminidase from Mucor hiemalis2008

    • Author(s)
      梅川碧里, ら
    • Organizer
      24th International Carbohydrate Symposium
    • Place of Presentation
      オスロ、ノルウェー国
    • Year and Date
      2008-07-29
    • Related Report
      2010 Final Research Report
  • [Presentation] Simple Synthesis of Influenza Virus Binding-Inhibitor Composed of Only Sugar Chains from Natural Compounds2008

    • Author(s)
      梅村舞子, ら
    • Organizer
      24th International Carbohydrate Symposium
    • Place of Presentation
      オスロ、ノルウェー国
    • Year and Date
      2008-07-29
    • Related Report
      2010 Final Research Report
  • [Presentation] Simple Synthesis of Influenza Virus Binding-Inhibitor Composed of Only Sugar Chains from Natural Compounds2008

    • Author(s)
      M. Umemura, et al.
    • Organizer
      24th International Carbohydrate Symposium
    • Place of Presentation
      Oslo, Norway
    • Year and Date
      2008-07-29
    • Related Report
      2008 Annual Research Report
  • [Presentation] Development of a Novel Glycosynthase Mutant of Endo-β-N-acetylglucosaminidase from Mucor hiemalis2008

    • Author(s)
      M. Umekawa, et al.
    • Organizer
      24th International Carbohydrate Symposium
    • Place of Presentation
      Oslo, Norway
    • Year and Date
      2008-07-29
    • Related Report
      2008 Annual Research Report
  • [Book] 糖鎖化学の基礎と実用化 エンド型グリコシダーゼを用いた糖ペプチドの化学-酵素合成2010

    • Author(s)
      山本憲二
    • Publisher
      シーエムシー出版
    • Related Report
      2010 Annual Research Report 2010 Final Research Report
  • [Book] 糖鎖を知る ひとつだけの微生物酵素を用いて糖鎖を付ける方法2010

    • Author(s)
      山本憲二
    • Publisher
      科学技術振興機構
    • Related Report
      2010 Annual Research Report 2010 Final Research Report
  • [Book] 複合糖質の化学と最新応用技術2009

    • Author(s)
      梅川碧里、山本憲二
    • Publisher
      シーエムシー出版
    • Related Report
      2010 Final Research Report
  • [Book] 複合糖質の化学と最新応用技術2009

    • Author(s)
      梅川碧里, 山本憲二(分担執筆)
    • Total Pages
      8
    • Publisher
      シーエムシー出版
    • Related Report
      2009 Annual Research Report
  • [Remarks] ホームページ

    • URL

      http://www.lif.kyoto-u.ac.jp/labs/molecule/ammbio/index.html

    • Related Report
      2010 Final Research Report
  • [Remarks]

    • URL

      http://www.ishikawa-pu.ac.jp/index.html

    • Related Report
      2010 Annual Research Report

URL: 

Published: 2008-04-01   Modified: 2016-04-21  

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