| Project/Area Number |
07556080
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
植物保護
|
|---|
| Research Institution | KYOTO UNIVERSITY |
|---|
Principal Investigator |
FURUSAWA Iwao Kyoto University, Faculty of Agriculture, Professor, 農学研究科, 教授 (10026594)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NAKANE Hiroyuki Toyota Motor Corporation, 研究部・バイオラボ, 研究員
OBATA Syuusei Toyota Motor Corporation, 研究部・バイオラボ, 研究員
KUBO Yasuyuki Kyoto Prefectural University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (80183797)
|
|---|
| Project Period (FY) |
1995 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1997: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1996: ¥3,000,000 (Direct Cost: ¥3,000,000)
|
|---|
| Keywords | Melanin / Carpropamid / Scytalone / Colletotrichum lagenarium / Magnaporthe grisea / Trihydroxynaphthalene / Appressoria / Magnaporthe grisea / トリヒドロオキシナフタレン / メラニン合成酵素 / サイタロン脱水酵素遺伝子 / p-ジフェノールオキシダーゼ / 酵素精製 / 結晶解析 |
|---|
| Research Abstract |
Melanin biosynthesis is essential for appressorial penetration of Colletotrichum lagenarium. Three major melanin biosynthetic enzymes, polyketide synthase PKS1, scytalone dehydratase SCD1 and 1,3,8-trihydroxynaphthalene reductase THR1 were expressed in E.coli and purified. We used either pMAL and pET system for the heterologous expression of proteins and purified by affinity chromatography. And in the case of PKS1, we constructed an expression vector for Aspergillus oryzae and identified the products by PKS1. We analyzed the mode of action of an novel anti-rice blast fungicide carpropamid by using purified enzymes. First, we established in vitro reaction system and then analyzed the inhibitory activity against the dehydration reaction by carpropamid. The result obtained was that carpropamid directly act to the enzyme and thus bringing about inhibition of melanin biosynthesis. Further we analyzed multimer formation of SCD1 by yeast two-hybrid system. The result obtained indicated that SCD1 enzyme associates to each other and would form multimer. Lastly we prepared polyclonal antibody against purified PKS1, SCD1 and THR1 and immuno-blot analysis of those melanin biosynthetic enzymes indicated that there would be a kind of post-translational regulation for the enzyme activity.
|
