| Project/Area Number |
14572008
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemical pharmacy
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| Research Institution | Nagoya City University, Graduate School of Pharmaceutical Sciences |
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Principal Investigator |
MIZUKAMI Hajime Nagoya City University, Graduate School of Pharmaceutical Sciences, Professor, 大学院・薬学研究科, 教授 (30128219)
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| Co-Investigator(Kenkyū-buntansha) |
NAGATSU Akito Nagoya City University, Graduate School of Pharmaceutical Sciences, Assistant Professor, 大学院・薬学研究科, 講師 (70244572)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Plant cell culture / Biotransformation / Glucosylation / Curcumin / UDP-glucosyltransferase / cDNA cloning / Recombinant enzyme |
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| Research Abstract |
We found that Catharanthus roseus cell suspension cultures converted exogenously supplied curcumin to a series of glucosides, none of which has been found in nature so far. The efficiency of glucosylation was dependent on culture stage of the cells and medium sucrose concentration. Methyl jasmonate and salicylic acid enhanced the glucosides formation only when they were added to the cultures prior to the addition of curcumin. The glucoside yield was 2.5 μmol/g fresh weight of the cells at an optimal culture condition. The water solubility of curcumin 4',4"-O-b-D-gentiobioside was 0.65 mmol/ml, which was 20 million-fold higher than that of curcumin.
Then, we isolated two cDNAs two cDNAs encoding UDP-glucosyltransferases (CaUGT1 and CaUGT2) from a CDNA library of cultured C roseus cells, using a PCR method directed at the UDP-binding domain of plant glycosyltransferases. We further confirmed that the recombinant CaUGT2 protein expressed in E. coli catalyzed the formation of curcumin monoglucoside from curcumin and also conversion of curcumin monoglucoside to curcumn diglucoside. It was shown that the use of the recombinant CaUGT2 may provide a useful new route for the production of culcumin glucosides.
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