Studies of the low-cost immobilization system for recombinant proteins using beta-GRP and curdlan.

• Project/Area Number: 24560954
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Health Sciences University of Hokkaido (2013-2014); Hokkaido University (2012)
• Principal Investigator: HORIUCHI Masataka 北海道医療大学, 薬学部, 講師 (90322825)
• Co-Investigator(Kenkyū-buntansha): 落合 正則 北海道大学, 低温科学研究所, 准教授 (10241382)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: タンパク質 / バイオテクノロジー / バイオリアクター / 生物・生体工学 / ナノ材料

Outline of Final Research Achievements

Amino terminal domain of Silkworm βGRP (GRP-tag) tightly and specifically associates with a water-insoluble β-1,3-glucan, curdlan. We have studied a novel immobilization system for recombinant proteins based on the association between the GRP-tag and curdlan. The recombinant proteins fused to the GRP-tag were expressed in several hosts and were immobilized in one step on the curdlan beads from cell lysates and medium. Furthermore, we succeeded in preparation of a curdlan disk and curdlan sheet as a support carrier for GRP-tag.

Research Products

• [Journal Article] The N-terminal domain of TIR domain-containing adaptor molecule-1, TICAM-1. (2014)
  • Author(s): Kumeta H, Sakakibara H, Enokizono Y, Ogura K, Horiuchi M, Matsumoto M, Seya T, Inagaki F.
  • Journal Title: J Biomol NMR Volume: 58（3） Issue: 3 Pages: 227-30
  • DOI: 10.1007/s10858-014-9819-1
  • Peer Reviewed
• [Journal Article] SuperNova, a monomeric photosensitizing fluorescent protein for chromophore-assisted light inactivation (2013)
  • Author(s): K. Takemoto (1), T. Matsuda (2)., 他12名
  • Journal Title: Sci. Rep Volume: 3巻 Issue: 1 Pages: 2629-2629
  • DOI: 10.1038/srep02629
  • NAID: 120006995014
  • Peer Reviewed
• [Journal Article] Structures and interface mapping of the TIR domain-containing adaptor molecules involved in interferon signaling. (2013)
  • Author(s): Enokizono Y, Kumeta H, Funami K, Horiuchi M, Sarmiento J, Yamashita K, Standley DM, Matsumoto M, Seya T, Inagaki F.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 110(49) Issue: 49 Pages: 19908-19913
  • DOI: 10.1073/pnas.1222811110
  • Peer Reviewed
• [Journal Article] A low-cost affinity purification system using beta-1,3-glucan recognition protein and curdlan beads (2012)
  • Author(s): Horiuchi, M., Takahasi, K., Kobashigawa, Y., Ochiai, M. and Inagaki, F.
  • Journal Title: PEDS Volume: 25 Issue: 8 Pages: 405-413
  • DOI: 10.1093/protein/gzs028
  • Peer Reviewed
• [Presentation] オッセオインテグレーション獲得に関わるタンパク質の昆虫細胞を用いた生産 (2014)
  • Author(s): 堀内留美，堀内正隆，横山敦郎
  • Organizer: 平成26年度 日本補綴歯科学会 東北北海道支部学術大会
  • Place of Presentation: 市民交流プラザ (郡山市)
  • Year and Date: 2014-10-25 – 2014-10-26
• [Presentation] Investigation of the inhibitory factors for osseointegration in the early stage of the implantation. (2014)
  • Author(s): Horiuchi M, Horiuchi R, Ochiai M, Yokoyama A
  • Organizer: The 28th Annual Symposium of the Protein Society
  • Place of Presentation: Manchester Grand Hyatt (San Diego, USA)
  • Year and Date: 2014-07-27 – 2014-07-30