| Project/Area Number |
18K19191
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 38:Agricultural chemistry and related fields
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| Research Institution | Kwansei Gakuin University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
柳原 格 地方独立行政法人大阪府立病院機構大阪母子医療センター(研究所), 免疫部門, 部長 (60314415)
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| Project Period (FY) |
2018-06-29 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
Fiscal Year 2019: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2018: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
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| Keywords | 分子シャペロニン / アーキア / 酢酸菌 / 超好熱菌 / 酵素の安定化 / タンパク質安定化 / アーケア / 安定化 |
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| Outline of Final Research Achievements |
The target recognition region of the hyperthermophilic molecular chaperonin is negatively charged, and it is considered that it preferentially captures basic molecules by electrostatic interaction in addition to hydrophobic one. The cytoplasmic pH of acetic acid bacteria was lower than that of Escherichia coli. It was expected that foreign protein molecules with a high isoelectric point are preferentially captured by the hyperthermophilic molecule chaperonin in the cells of acetic acid bacteria. Using β-galactosidase of E. coli as a model target, the effect was evaluated in the presence or absence of hyperthermophilic chaperonin in Komagataeibacter europaeus. β-galactosidase was significantly stabilized by coexpression of the molecular chaperonin CpkB of the hyperthermophilic archaeon Thermococcus kodakarensis. In addition, in vitro effect was also examined for various proteins, showing that the effect was effective for a target having a smaller molecular weight.
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| Academic Significance and Societal Importance of the Research Achievements |
酵素の安定化は、分子を効率的に精製するうえで重要である。対象となる酵素を選択的に安定化し、それ以外の分子を除去できれば、精製は用意になる。本研究では超好熱菌の分子シャペロニンの標的認識機構に注目し、低pH環境で選択的安定化がなされるか検証した。特に酢酸菌を宿主として組み合わせることで、目的とする分子を優先的に分子シャペロニンに捕捉させ、単純な操作で分画できれば、精製は効率化される。酢酸菌は食酢生産に用いられてきた微生物であり、安全性も高い。抗生物質を加えなくても雑菌繁殖を抑えることができる。本方法を発展させることで、安全で効率的な物質生産用宿主ベクター系が期待できる。
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