| Project/Area Number |
16H03282
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Biomolecular chemistry
|
|---|
| Research Institution | Kyoto University |
|---|
Principal Investigator |
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
水谷 公彦 京都大学, 農学研究科, 助教 (40314281)
|
|---|
| Project Period (FY) |
2016-04-01 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000)
Fiscal Year 2018: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2017: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2016: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
|
|---|
| Keywords | 生合成 / プレニル化 / ポリフェノール / 膜結合型酵素 / 酵素 / 植物 / 蛋白質 / 生体分子 / 蛋白 |
|---|
| Outline of Final Research Achievements |
Prenylated polyphenols are intensively studied as bio-active compounds in pharmaceutical and agricultural fields, and thus far ca. 1,000 compounds are known. Prenyltransferases involved in those natural compounds are newly identified protein family, and due to the membrane-bound property the catalytic mechanisms are yet largely unknown. Toward crystallization of this enzyme family, this study aimed the high production of these enzymes in heterologous hosts as budding yeast, taking 29 genes isolated from 8 different plant families, to which GFP-His tag was attached both at N- and C-terminal. The resulting proteins were evaluated at protein expression level, solubilization, and affinity purification, resulting in 9 clones showed GFP fluorescence, 4 clones were solubilized, and two was successfully purified, while their protein amounts was still low for crystallization.
In parallel, 3D modeling was done based on our biochemical data and archeae prenyltransferase crystal structure.
|
| Academic Significance and Societal Importance of the Research Achievements |
プレニル化フェノールは、薬学、農学、食品科学の分野で注目される天然有機化合物で、多くは薬用植物から単離同定され、これまでに約1,000種の構造が明らかになっている。一方、その生合成のカギを握るプレニル基転移酵素は、膜結合型でその全体像には不明な点が多い。そこで本研究では、８科に渡る植物から２９種の同酵素遺伝子を単離し、大量発現を試みた。この酵素の機能を分子レベルで知ることは、人の健康にメリットのある天然物の安定供給につながると期待される。
|
