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Challenge for production of therapic glyco-peptides using gene expression system in wood rot fungi

Research Project

Project/Area Number 25660137
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Wood science
Research InstitutionKyoto University

Principal Investigator

HONDA Yoichi  京都大学, (連合)農学研究科(研究院), 教授 (70252517)

Co-Investigator(Kenkyū-buntansha) WATANABE Takashi  京都大学, 生存圏研究所, 教授 (80201200)
Co-Investigator(Renkei-kenkyūsha) NAKAZAWA Takehito  京都大学, 大学院農学研究科, 助教 (80608141)
YOSHIOKA Koichi  京都府立大学, 大学院生命環境科学研究科, 特任助教 (20540149)
Research Collaborator NAGAOKA Jun  京都大学, 大学院農学研究科
ITO Keigo  京都大学, 農学部森林科学科
Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywords木材腐朽菌 / 糖タンパク質 / 異種発現 / 糖鎖工学 / 担子菌 / 糖ペプチド
Outline of Final Research Achievements

It was demonstrated that wild type MnP2 enzyme from Pleurotus ostreatus contains a GlcNac2-Hex5 glycan attached to Asp102 using MALDI TOF-MS analysis. This is the first report of glycan structure attached to fungal lignin-degrading peroxidase. We also tried to analyze an additional glycan supposed to its artificially modified mutant, MnP2 R263N, but production and purification of the enzyme was not enough for the carbohydrate analysis. Moreover, a cDNA fragment encoding for cholera toxin B subunit was inserted to an expression vector for Pleurotus ostreatus and tried to introduced into host strain but no protein-producing transformants have been isolated so far.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • Research Products

    (4 results)

All 2014 2013

All Presentation (4 results) (of which Invited: 1 results)

  • [Presentation] Analysis of N-glycan of the extracellular versatile peroxidase MnP2 secreted by Pleurotus ostreatus2014

    • Author(s)
      Keigo ITO, Jun NAGAOKA, Koichi YOSHIOKA, Takehito NAKAZAWA, Masahiro SAKAMOTO, Takashi WATANABE & Yoichi HONDA
    • Organizer
      8th International Conference on Mushroom Biology and Mushroom Products
    • Place of Presentation
      New Delhi, India
    • Year and Date
      2014-11-19 – 2014-11-22
    • Related Report
      2014 Annual Research Report
  • [Presentation] Genetic transformation of mushrooms and its utilization in basic and applied sciences2014

    • Author(s)
      Yoichi Honda
    • Organizer
      8th International Conference on Mushroom Biology and Mushroom Products
    • Place of Presentation
      New Delhi, India
    • Year and Date
      2014-11-19 – 2014-11-22
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] ヒラタケの野生型MnP2の糖鎖構造解析2014

    • Author(s)
      伊藤奎梧, 長岡潤, 吉岡康一, 中沢威人, 坂本正弘, 渡辺隆司, 本田与一
    • Organizer
      日本きのこ学会25周年記念大会
    • Place of Presentation
      京都
    • Year and Date
      2014-09-10 – 2014-09-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] 白色腐朽菌の生産する糖タンパク質のN-結合型糖鎖と外来タンパク質の修飾2013

    • Author(s)
      長岡潤、伊藤奎梧、中沢威人、坂本正弘、 本田与一、吉岡康一、渡辺隆司
    • Organizer
      シンポジウム「微生物科学研究の多様性と新展開」
    • Place of Presentation
      京都大学
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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