| Project/Area Number |
17K15246
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied microbiology
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| Research Institution | Kyushu University |
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Principal Investigator |
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| Research Collaborator |
SUEMATSU Yuri
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| Project Period (FY) |
2017-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | silica / thermostable / シリカ / 耐熱化 / 異種発現系 / expression / バイオシリカ |
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| Outline of Final Research Achievements |
Silicatein and silicase are unique enzymes which catalyze the polymerazation and degradation between silicic acids and silica, respectively. Random mutations were introduced into silicatein gene and then thermostable (>70oC) protein was investigated. We found one candidate mutant by first screening, but it turned to be negative by second screening. We tried the expression system with Brevibacillus to obtain soluble protein in medium fraction, however, no activity was observed with expressed proteins. On the other hand, expression vector in thermophilic bacterium, Thermus thermophilus, was developed. The reporter assay indicated high expression of target protein and the yield reached 28 mg protein per 1L culture. This expression system can be applied in industrial use.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では目的とした耐熱性を持ったシリカ重合酵素を作製することは出来なかったが、その発現方法や活性について知見を蓄積することができた。今後、より可溶性が高く耐熱性が高いタンパク質をつなげて活性評価を行うことで目的が達成できると思われる。また、今回作製することのできた好熱菌を宿主とするタンパク質発現系は、従来に比べて発現量が多く、産業利用にも期待が持てる。耐熱性酵素は安定性が高いため、様々なニーズがあるが、本システムを用いることによりそれらが産業に利用できるだけでなく、今回行ったような耐熱化酵素研究の宿主とすることができ、より研究を加速するのに寄与できると思われる。
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