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2007 Fiscal Year Final Research Report Summary

Proteomic Analysis of Extracellular Enzymes produced by the Basidiomycetes Fungi during Cellulose Degradation

Research Project

Project/Area Number 17380102
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 林産科学・木質工学
Research InstitutionThe University of Tokyo

Principal Investigator

SAMEJIMA Masahiro  The University of Tokyo, Graduate School of Agricultural and Life Sciences, professor (30162530)

Co-Investigator(Kenkyū-buntansha) KIYOHIKO Igarashi  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Assistant professor (80345181)
Project Period (FY) 2005 – 2007
KeywordsGenome / Glycoside Hydrolase / Biomass / Proteome / Cellulose / Basidiomycetes / Phanerochaete chrysosporium / Wood Rotting Fungi
Research Abstract

In this project, proteomic analysis of extracellular enzymes produced by the Basidiomycete Phanerochaete chrysosporium during cellulose degradation was performed. Two-dimensional gel electrophoresis for extracellular solution, LS-MS/MS analysis for each enzyme separated on the gel to identify the partial sequence of amino acids, and searching on the P.chrysosporium database to find the corresponding genome sequences were applied for this purpose. After these processes, the cDNAs of the target enzymes depending on the purposes desired were cloned and transformed in the yeast Pichia pastoris with expression vectors to produce the recombinant enzymes of them.
On the basis of these techniques established, we have succeeded to obtain several cellulases and related enzymes produced during cellulose degradation by this fungus as recombinant enzymes. Among them, an endoglucanase in GH family 45 and a xyloglucanaase in GH family 74 were identified as novel enzymes and well characterized by this project. In addition, the novel beta-1, 3-glucanase in GH family 16, named Lam 16A, has been found during cellulose degradation by this fungus. As a collaboration work with the research group in the Swedish University of Agricultural and Life Sciences, functional and structural analysis of Lam16A was accomplished in a part of this project. Moreover, we have also succeeded to synthesis the novel cycloglucan using a site-directed mutant enzyme derived from Lam16.
In conclusion, we have established the total technical system from proteomic analysis of extracellular solution of the basidiomycete P.chrysosporium to production of recombinant enzymes for the conversion of cellulosic biomass.

  • Research Products

    (15 results)

All 2010 2009 2008 2007 2006

All Journal Article (12 results) (of which Peer Reviewed: 6 results) Presentation (1 results) Patent(Industrial Property Rights) (2 results)

  • [Journal Article] Synthesis of Cyclic β-Glucan Using Laminarinase 16A Glycosynthase Mutant from the Basidiomycete Phanerochaete chrysosporium2010

    • Author(s)
      J.Vasur, R.Kawai, K.H.M.Jonssou, G.Widmalm, A.Engstrm, M.Frank, E.Andersson, H.Hansson, Z.Farsberg, K.Igarashi, M.Samejima, M.Sandgren, J.Stahlberg
    • Journal Title

      J.Am.Chem.Soc. 132

      Pages: 1724-1730

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Synthesis of Cyclic β-Glucan Using Laminarinase 16A Glycosynthase Mutant from the Basidiomycete Phanerochaete chrysosporium2010

    • Author(s)
      J.Vasur, R.Kawai, K.H.M.Jonsson, G.Widmalm, A.Engstrm, M.Frank, E.Andersson, H.Hansson, Z.Forsberg, K.Igarashi, M.Samejima, M.Sandgren, J.Stahlberg
    • Journal Title

      J.Am.Chem.Soc. 132

      Pages: 1724-1730

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] X-ray crystal structures of Phanerochaete chrysosporium Laminarinase 16A in complex with products from lichenin and laminarin hydrolysis2009

    • Author(s)
      J.Uasur, R.Kawai, E.Andersson, K.lgarashi, M.Sandgren, M.Samejima, J.Stahlberg:
    • Journal Title

      FEBS J 276

      Pages: 3858-3869

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] X-ray crystal structures of Phanerochaete chrysosporium Laminarinase 16A in complex with products from lichenin and laminarin hydrolysis2009

    • Author(s)
      J.Vasur, R.Kawai, E.Andersson, K.Igarashi, M.Sandgren, M.Samejima, J.Stahlberg
    • Journal Title

      FEBS J. 276

      Pages: 3858-3869

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Characterization of an endoglucanase belonging to a new subfamily of glycoside hydrolase family 45 of the basidiomycete Phanerochaete chrysosporium2008

    • Author(s)
      K.Igarashi, T.Ishida, .C Hori, M.Samejima
    • Journal Title

      Appl. Environ. Microbiol 74

      Pages: 5628-5634

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Characterization of an endoglucanase belonging to a new subfamily of glycoside hydrolase family 45 of the basidiomycete Phanerochaete chrysosporium2008

    • Author(s)
      K.Igarashi, T.Ishida, C.Hori, M.Samejima
    • Journal Title

      Appl.Environ.Microbiol. 74

      Pages: 5628-5634

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Substrate recognition by glycoside hydrolase family 74 xyloglucanase from the basidiomycete Phanerochaete chrysosporium2007

    • Author(s)
      T.Ishida K.Yaoi, A.Hiyoshi, K.Igarashi, M.Samejima
    • Journal Title

      FEBS J. 274

      Pages: 5727-5736

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Substrate recognition by glycoside hydrolase family 74 xyloglucanase from the basidiomycete Phanerochaete chrysosporium2007

    • Author(s)
      T.Ishida, K.Yaoi, A.Hiyoshi, K.Igarashi, M.Samejima
    • Journal Title

      FEBS J. 274

      Pages: 5727-5736

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Hydrolysis of beta- 1,3/1,6-glucan by glycoside hydrolase family 16 endo-1,3(4)-beta-glucanase from the basidiomycete Phanerochaete chrysosporium2006

    • Author(s)
      R.Kawai, K.Igarashi, M.Yoshida, M.Kitaoka, M.Samejima
    • Journal Title

      Appl. Microbiol. Biotechnol. 71

      Pages: 898-906

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] X-ray crystallographic native sulfur SAD structure determination of laminarinase Lam 16A from Phanerochaete chrysosporiu2006

    • Author(s)
      J.Vasur, R.Kawai, A.M.Larsson, K.Igarashi, M.Sandgren, M.Samejima, J.Stahlberg
    • Journal Title

      Acta Crystallographica Section D 62

      Pages: 1422-1429

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Hydrolysis of beta-1,3/1,6-glucan by glycoside hydrolase family 16 endo-1,3(4)-beta-glucanase from the basidiomycete Phanerochaete chrysosporium2006

    • Author(s)
      R.Kawai, K.Igarashi, M.Yoshida, M.Kitaoka, M.Samejima
    • Journal Title

      Appl.Microbiol.Biotechnol. 71

      Pages: 898-906

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] X-ray crystallographic native sulfur SAD structure determination of laminarinase Lam 16A from Phanerochaete chrysosporium2006

    • Author(s)
      J.Vasur, R.Kawai, A.M.Larsson, K.Igarashi, M.Sandgren, M.Samejima, J.Stahlberg
    • Journal Title

      Acta Crystallographica Section D 62

      Pages: 1422-1429

    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Cloning and Characterization of Encoding Glycoside Hydrolase Family 5 Enzymes from Basidiomycete Phanerochaete chrysosporium2008

    • Author(s)
      M.Okamoto, T.Ishida, M.Hiraishi, H.Suzuki, T.Tsukada, K.Igarashi, M.Samejima
    • Organizer
      Symposium on Wood Science and Technology 2008(IAWPS2008)
    • Place of Presentation
      Harbin, China
    • Year and Date
      20080927-20080929
    • Description
      「研究成果報告書概要(欧文)」より
  • [Patent(Industrial Property Rights)] バイオマス原料の処理方法,並びに糖の製造方法,エタノールの製造方法,及び乳酸の製造方法2009

    • Inventor(s)
      五十嵐圭日子、鮫島正浩
    • Industrial Property Rights Holder
      五十嵐圭日子、鮫島正浩
    • Industrial Property Number
      特願2009-146466
    • Filing Date
      2009-06-19
    • Description
      「研究成果報告書概要(和文)」より
  • [Patent(Industrial Property Rights)] エンドグルカナーゼ活性を有する新規タンパク質、そのDNA、及びそれらの利用2008

    • Inventor(s)
      五十嵐圭日子、鮫島正浩
    • Industrial Property Rights Holder
      五十嵐圭日子、鮫島正浩
    • Industrial Property Number
      特願2008-185219
    • Filing Date
      2008-07-16
    • Description
      「研究成果報告書概要(和文)」より

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Published: 2011-06-18  

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