| Project/Area Number |
17360393
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TSUMOTO Kouhei The University of Tokyo, Graduate School of Frontier Sciences, Associate Professor (90271866)
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| Co-Investigator(Kenkyū-buntansha) |
UMETSU Mitsuo Tohoku University, Graduate School of Engineering, Associate Professor (70333846)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥16,430,000 (Direct Cost: ¥15,500,000、Indirect Cost: ¥930,000)
Fiscal Year 2007: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2006: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2005: ¥8,300,000 (Direct Cost: ¥8,300,000)
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| Keywords | PROTEIN / BIOTECHNOLOGY / BIOMOLECULE / BIO-ENGINEERING / 溶媒・添加剤 / アルギニン / 可溶化 / クロマトグラフィー / 凝集会合形成 / フィロールディング / 製剤 / プロテインマニピュレーション / リフォールディング |
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| Research Abstract |
To establish the novel system for solublization and preparation of proteins using amino acids and their derivatives, we have focused on the characteristics of arginine and it derivatives; mechanism on protein aggregation suppression of arignine and its derivatives and its application to several bioprocesses for proteins have been addressed from the following viewpoints.
1. Elucidation of aggregation suppression mechanism on proteins of L-arginine and its derivatives Solubility measurements of L arginine and its derivatives with various amino acids, and peptide bonds have indicated the identical affinities for main chain and side chains to those of guanidium ion. Several derivatives could improve the specific characteristics of L-arginine, e.g. efficiency of solubilization of proteins. 2) Crystal structures of two proteins under existence of L-arginine: crystal structures of two proteins have indicated that L-arginine made critical contribution to the stabilization of hydrated proteins,
… More
and also suggested that the mechanism on the proteins is dependent upon the tertiary structures of target proteins.
2. Solubilization of proteins using L-arginine and its derivatives E-coli expressed signal-transduction-related proteins, and membrane proteins could be solubilized from inclusion bodies by using L-arginine and its derivatives. Thermodynamics analyses have revealed the complete recovery of the functions and structures of the solubilized proteins. We could successfully establish the method for solubilization of proteins using L-arginine and its derivatives.
3. Application of arigine to solvent additives of chromatographies: it could be shown that arginine could be utilized for an effective solvent additive of various chromatographies, including GPC, HIC, IEX, and several affinity chromatographies.
From these results, our achievements could be summarized as follows; the mechanism of arginine on protein structure, effective utilization of arginine for solubilzation and purification of proteins. Less
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