| Project/Area Number |
15K07366
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Kyoto Prefectural University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
川崎 一則 国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (40356837)
増村 威宏 京都府立大学, 生命環境科学研究科, 教授 (50254321)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Keywords | 膜小胞 / プロテアーゼ / 巨大分子複合体 / 表面提示 / ジスルフィドオキシダーゼ / 温度感受性 / 好熱性細菌 |
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| Outline of Final Research Achievements |
Macromolecular complexes of keratinolytic proteases and membrane vesicles produced by Meiothermus ruber H328 were employed for the study. A protease responsible for keratin hydrolysis of chicken feathers was identified by combining the proteome analysis with different preparations of strain H328 with the whole genome analysis. The protease was confirmed to hold strong resistance toward various denaturants including SDS and organic solvents. This resistance presumably occurred due to thermophilic properties and membrane vesicles. A simple method including filtration and ultracentrifugation for the culture supernatant was developed by use of a fluorescent dye, DiI. A degP-homologue candidate was selected and its knockout mutant was constructed. Production of membrane vesicles was enhanced at an elevated temperature 60°C. To develop the system to display a protein on the surface of membrane vesicles, three candidates were screened by use of a software searching membrane proteins.
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