| Project/Area Number |
17510188
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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 |
Living organism molecular science
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
MURANAKA Toshiya The Institute of Physical and Chemical Research, Metabolic Diversity Research Team, Team Leader (60342862)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Hiroyuki Kyushu University, Pharmaceutical Sciences, Associate Professor (30253470)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2005: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | licorice / glycyrrhizin / hairy root / triterpene / P450 / oxygenase / glycosyltranferase / immunochemistry |
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| Research Abstract |
The roots and stolons of Glycyrrhiza plants contain a large amount of glycyrrhizin, an oleanane-type triterpene saponin. Various pharmaceutical researches for this compounds have been done, however, biosynthetic pathway from β-amyryin, a common triterpene in plants to glycyrrhizin, a final product has not been identified. In this study, we investigated on the metabolic pathway by the integration of transcriptome and immunochemical analysis using root culture systems. By using expressed sequence tag (EST) of cDNA library from stolons of G. uralensis plants, we mined putative cytochrome P450 monooxygenases (P450s). Subsequently, transcript profiling-based selection narrowed down to seven candidate P450s. Furthermore, we examined immunochemical assays using glycyrrhizin monoclonal antibody and Eastern blotting established for analysis of glycyrrhizin. After crude separation of the extract from the plant by TLC, compounds were transferred from the TLC plate to a PVDF membrane. Finally, we identified a new P450 catalyzing the sequential two-step oxidation at C-11 of β-amyrin to yield 11-oxo-β-amyrin.
The integration of transcriptome and immunochemical analysis using root culture systems is useful for studying plant metabolic pathways.
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