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Establishment of high concentration Refolding of unstable protein

Research Project

Project/Area Number 21580119
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied biochemistry
Research InstitutionSojo University

Principal Investigator

MATSUMOTO Toshihiko (2010-2011)  崇城大学, 生物生命学部, 准教授 (60133568)

井本 泰治 (2009)  Sojo University, 生物生命学部, 教授 (90038282)

Co-Investigator(Kenkyū-buntansha) MATSUMOTO Toshihiko  崇城大学, 生物生命学部, 准教授 (60133568)
YAMAURA Izumi  崇城大学, 生物生命学部, 教授 (30133565)
Project Period (FY) 2009 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2010: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords再生 / フォルディング / 安定化剤 / 可溶化剤 / 不安定蛋白質 / タンパク質の再生 / リゾチーム / グリセリン / 透析再生法 / 高濃度生成法 / 不安定タンパク質 / 高濃度再生法
Research Abstract

Earlier, we formally established an effective refolding procedure for a protein by gradient removal of a solubilizer such as urea. However, this procedure was less effective for unstable proteins. We developed here an excellent method to add protein stabilizer so as to get reasonable amounts of folded protein under the concentration of solubilizer where the unstable protein does not form aggregate. We examined many stabilizers and found that 60% of a concentrated(2.5 mg/ml) unstable protein can be refolded using 40% glycerol as the best stabilizer. This procedure can be widely applicable for the refolding of unstable proteins.

Report

(4 results)
  • 2011 Annual Research Report   Final Research Report ( PDF )
  • 2010 Annual Research Report
  • 2009 Annual Research Report
  • Research Products

    (4 results)

All 2012 2010

All Journal Article (3 results) (of which Peer Reviewed: 3 results) Presentation (1 results)

  • [Journal Article] Refolding of an unstable lysozyme by gradient removal of a solubilizer and gradient addition of a stabilizer2010

    • Author(s)
      Kohyama K, Matsumoto T, and Imoto T
    • Journal Title

      J. Biochem

      Volume: 143巻 Pages: 427-431

    • NAID

      10027874720

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Effect of protein concentration and pH on the chitinase activity of Tapes japonica lysozyme2010

    • Author(s)
      Goto T, Abea Y, Imoto T, Uedaa T
    • Journal Title

      Protein Peptide Let. 17

      Pages: 172-175

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Refolding of an unstable lysozyme by gradient removal of a solubilizer and gradient addition of a stabilizer2010

    • Author(s)
      Kohyama K, Matsumoto T, Imoto T
    • Journal Title

      J.Biochem. 143

      Pages: 427-431

    • NAID

      10027874720

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Presentation] 不安定タンパク質の効率的な再生法の確立2012

    • Author(s)
      保田達哉, 野稲俊宏, 松元俊彦, 安藤祥司, 井本泰治
    • Organizer
      日本農芸化学会
    • Place of Presentation
      京都女子大
    • Year and Date
      2012-03-24
    • Related Report
      2011 Annual Research Report 2011 Final Research Report

URL: 

Published: 2009-04-01   Modified: 2016-04-21  

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