| Project/Area Number |
06556013
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
OHTA Akinori The Univ.of Tokyo, Facul.of Agric., Assoc.Prof., 農学部, 助教授 (30125885)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUI Tohru Japan Energy Co., Instit.of Pharmacol.& Biotech., Investigator, 医薬バイオ研究所, 研究員
FURUHASHI Keizo Japan Energy Co., Instit.of Pharmacol.& Biotech., Chief Investigator, 医薬バイオ研究所, 主席研究員
NAGATA Yuji The Univ.of Tokyo, Facul.of Agric., Assis.Prof., 農学部, 助手 (30237531)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1994: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | Candida maltosa / Cytochrome P-450 / n-Alkane / Dicarboxylic Acid / Gene Disruption / Acyl-CoA Oxidase / Endoplasmic Reticulum / Peroxisome / チトクローム P-450 / 遺伝子工学 / 酵母 / チトクロームP450 / アシルCoA オキシダーゼ / Candida tropicalis |
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| Research Abstract |
The research plan constituted of following three parts. (1) Construction of new dicarboxylic acid-producing yeast by using genetic engineering system which was developed for n-alkane-assimilating yeast Candida maltosa in our laboratory. (2) Improvement of production efficiency and purity of the dicarboxylic acid by overproducing specific cytochrome P-450. (3) Improvement of dicarboxylic acid production by disruption of acyl-CoA oxidase gene which works in degradation of the product.
In 1994, we analyzed various genes encoding cytochromes P-450 in C.maltosa.and attained overproduction of specific cytochrome P-450 by GAL gene promoters. We also analyzed the substrate specificity and catalyzed products of the cytochromes. In 1995, disruption of C.maltosa POX2 gene encoding acyl-CoA oxidase was successfully accomplished. Production of dicarboxylic acid from n-tetradecane by the disrupted strain was subsequently confirmed by the Japan Energy group who had fermentation and detection equipments. We constructed a vector that was stably maintained in C.maltosa and isolated cytochrome P-450 reductase gene, of which overexpression will further improve alkane oxidation by C.maltosa.
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