Development of evaluation methods for enzyme activities for alkane biosynehtsis
| Project/Area Number |
26870136
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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 |
Chemical biology
Applied biochemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
HAYASHI Yuuki 東京大学, 総合文化研究科, 助教 (90444059)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Keywords | アルカン生合成 / 進化分子工学 / 蛋白質工学 / アシル-ACP還元酵素 / アルデヒド脱ホルミル化酸化酵素 / タンパク質 / アルカン合成酵素 / バイオエネルギー / バクテリアルシフェラーゼ / アシル輸送蛋白質還元酵素 / アルデヒド脱炭酸酵素 / アルカン生産 |
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| Outline of Final Research Achievements |
It is too low for activities of enzymes (AAR and AD), to produce alkane by microorganisms, practically. Therefore. in this study, we proposed that the enzyme activities were improved by directed evolution. Directed evolution is a method to improve and/or change characteristics and activities of enzymes, and require a high-throughtput method to evaluate AD and AAR activities of mutants, quickly and simply.
At first, using Nile red, we tried to detect amount of alkane produced by co-expression of AD and AAR as s in vivo AD activity, however, we did not discriminate activities among AD mutants. Thus, we needed to change a strategy to elucidate in vivo AD activity. On the other hand, we successfully established a method to evaluate in vivo AAR activity directly as bioluminescence by coexpressed bacterial luciferase. Using this method, we can simultaneously compare in vivo AAR activity among over 1000 AAR mutants, resulting in accelerating improvement of AAR activity by directed evolution.
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Report
(3 results)
Research Products
(29 results)
