| 研究課題/領域番号 |
22J20479
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| 配分区分 | 補助金 |
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| 研究機関 | 筑波大学 |
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研究代表者 |
ALAWI ASEEL 筑波大学, 理工情報生命学術院, 特別研究員(DC1)
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| 研究期間 (年度) |
2022-04-22 – 2025-03-31
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| キーワード | glycerol / 1,3-propanediol / 3-hydroxypropionic acid / Citrobacter braakii |
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| 研究実績の概要 |
To produce 3-hydroxypropionic acid (3-HP) from glycerol we used Citrobacter braakii strain TB-96-dLF, which was previously modified to produce high 1,3-propandiol (1,3-PD) concentration from glycerol. First, we disrupted propanediol dehydrogenase (pdh) gene to block 1,3-PD production (TB-96-dLFP). Since TB-96-dLFP may accumulate the toxic 3-hydroxypropionaldehyde (3-HPA) intermediate, we fermented it under aerobic condition as to produce 3-HP from glycerol via 3-HPA. However, no production of 3-HP was observed. This may be due to the instability of the first enzyme of glycerol metabolism which is glycerol dehydratase (Gdh) under aerobic conditions.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Even though we expected that the deletion of propanediol dehydrogenase (pdh) gene in strain TB-96-dLFP will result in 3-hydroxypropionic acid (3-HP) production from glycerol via 3-hydroxypropionaldehyde (3-HPA), fermentation under aerobic conditions showed low glycerol consumption and 3-HP production was not observed. This was explained by the instability of glycerol dehydratase (Gdh)the first enzyme involved in glycerol metabolism. Thus, further enhancement of gdh is necessary under aerobic conditions.
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| 今後の研究の推進方策 |
Aiming at the enhancement of glycerol metabolism under aerobic conditions, overexpression of glycerol dehydratase (gdh) gene is needed. Thus, we plan to screen suitable over expression vectors that can be transformed and replicated in TB-96-dLF. Next, genetic modification of antibiotic resistance cassette will be conducted for easier selection. Last, cloning of related genes and construction of insert will be conducted.
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