| Project/Area Number |
08456172
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SATO Fumihiko KYOTO UNIVERSITY, GRAD. SCH. AGRIC., PROFESSOR, 農学研究科, 教授 (10127087)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1998: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Molecular engineering / Isoquinoline alkaloid biosynthesis / O-methyltransferese / Eschscholtzia / alkaloid composition / scoulerine-9-O-methyltransferase / tetrahydrocolumbamine / テトラハイドロコロンバシン / モルフィナンアルカロイド / 形質転換 / Agrobacterium / パーティクルカン / モルフィキンアルカロイド / パーティクルガン / イソキノリンアルカロイド / モロフィナンアルカロイド / パーティクルガン法 / 生合成 |
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| Research Abstract |
The molecular engineering of isoquinoline alkaloid biosynthesis in plant cells was investigated. Using cauliflower mosaic virus 35S promoter with enhancer elements, high expression vectors for 3 OMTs of berberine biosyntheis were constructed and introduced into 3 different plants ; Coptis (homologous plant), Eschscholtzia (a plant which produces benzophenanthridine alkaloid derived from scoulerine, an intermediate for berberine) and Nicotiana tabacum (nicotine alkaloid plant which does not produce isoquinoline type alkaloid). Although transformation of Coptis cells was very difficult, at least transformants with SMT (scoulerine 9-0-methyltransferase) expression were established for all host plant species. Transformed Coptis cells showed increased SMT activity and modified alkaloid composition ; I.e. more berberine and less coptisine (different end product from scoulerine). This result suggest the modification of branching point enzyme (I.e. SMT) would be a useful method to modify the composition of alkaloid, whereas modification was limited. Transformed Eschscholtzia cells also showed clear SMT activity and modified metabolite composition ; high accumulation of tetrahydrocolumbamine, a product of SMT reaction. On the other hand, the transformants of tobacco did not show SMT activity, whereas SMT transcripts were detected. Effect of host plant species on molecular engineering of alkaloid biosynthesis is discussed.
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