Improvement of protein stability for production of biofuels
Project/Area Number |
21760638
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Research Category |
Grant-in-Aid for Young Scientists (B)
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Allocation Type | Single-year Grants |
Research Field |
Biofunction/Bioprocess
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Research Institution | Kobe University |
Principal Investigator |
TAMAKA Tsutomu Kobe University, 自然科学系・先端融合研究環重点研究部, 助教 (90436551)
|
Project Period (FY) |
2009 – 2010
|
Project Status |
Completed (Fiscal Year 2010)
|
Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2010: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2009: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
|
Keywords | タンパク質 / 酵素 / 熱安定性 |
Research Abstract |
The aim of this study is to establish enzyme-mediated protein stabilization and its application inside living cells. We have focused on Sortase, a transpeptidase. Sortase recognizes LPXTG amino acid sequences and cleaves between T and G, and then ligated with N-terminal polyglycine sequences. Our strategy is that the substrate sequences, i.e., LPXTG and GGG sequences, are introduced both N- and C-terminus of the target protein. Then Sortase allows to conjugate its N- and C-terminus and produces the circular protein. EGFP were chosen as model proteins and these substrate tags appended EGFP was successfully conjugated by Soprtase and circular EGFP was obtained. Then the thermal stability of these circular proteins was significantly improved, suggesting our strategy is very useful for protein stabilization. The optimal linker design between N-terminal and C-terminal is one of the issue for expanding versatility of this method.
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Report
(3 results)
Research Products
(15 results)
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[Journal Article] Site-specific tetrameric streptavidin-protein conjugation using sortase A.2011
Author(s)
Matsumoto, T., Sawamoto, S., Sakamoto, T., Tanaka T., Fukuda, H., Kondo, A.
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Journal Title
Journal of Biotechnology 151(1-2)
Pages: 37-42
Related Report
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[Journal Article] Site-specific tetrameric streptavidin-protein conjugation using sortase A.2011
Author(s)
Matsumoto, T., Sawamoto, S., Sakamoto, T., Tanaka, T., Fukuda, H., Kondo, A.
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Journal Title
Journal of Biotechnology
Volume: 152
Pages: 37-42
Related Report
Peer Reviewed
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