Studies on Biosynthesis of Ecdysteroids with Plant Tissue Culture

植物组织培养脱皮素生物合成研究

• 批准号: 02640422
• 负责人: FUJIMOTO Yoshinori
• 金额: $ 1.34万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02640422/
• 关键词: ecdysteroid; 20-hydroxyecdysone; plant tissue culture; stable isotope; ^2H-NMR; biosynthetic pathway; 生合成機構; 安定同位体標識化合物; 毛状根; Ajuga reptans; 生合成メカニズム

1. Feeding experiment of (2- ^<13>C) acetate with the tissue culture of Ajuga hairy root revealed that it was incorporated into ecdysteroids efficiently. It was found that the 24-alkyl group of C-28 and C-29 ecdysteroids was not derived from acetate.2. Feeding of synthetic (26, 27- ^<13>C_2) cholesterol as an emulsion into Tween80 to the tissue culture was allowed to incorporate into 20-hydroxy-ecdysone efficiently, but not into C-28 and C-29 ecdysteroids.3. More importantly, the above experiment has demonstrated that stable isotopes such as ^1H and ^<13>C can be utilized in biosynthetic studies of ecdysteroids.4. (3 - ^2H)- and (4 - ^2H) Cholesterols were chemically synthesized.5. ^2H-NMR analysis of 20-hydroxyecdysones derived from the ^2H-labeled cholesterols revealed that the 3alpha- and 4beta-hydrogens were incorporated into 20-hydroxyecdysone without change of the location and stereochemistry.6.5beta- ^2H derivative of an intermediate, 3beta-hydroxy-5beta-cholest-7-en-6-one, which was postulated by the aforementioned results, was synthesized and fed with the culture system. The feeding experiment demonstrated that the intermediate was metabolized into 20-hydroxyecdysone much better than cholesterol.7. We now propose 3beta-hydroxy-5beta-cholest-7-en-6-one as a key intermediate in the earlier stage of ecdysone biosynthesis. Our present studies are focused on the identification of a precursor of this intermediate.

1。乙酸盐的（2- ^<13>）乙酸盐的喂养实验，其组织的组织培养物表明，它有效地掺入了ecdystroids。已经发现，C-28和C-29雌激素的24-烷基群未源自乙酸。2。将合成（26，27- ^<13> c_2）胆固醇作为乳液喂入tween80到组织培养物中，可以有效地掺入20-羟基 - Ecdysone中，但不能掺入C-28和C-28和C-29 ecdysteroids.3。更重要的是，以上实验表明，稳定的同位素（例如 ^1H和 ^<13> c）可以用于ecdysteroid.4。 （3 - ^2H） - 和（4- ^2H）胆固醇是化学合成的。5。从 ^2H标记的胆固醇衍生的20-羟基甲虫的 ^2H-NMR分析表明，将3Alpha-和4beta-Hydrogens纳入了20-羟基甲虫中3beta-Hydroxy-5beta-cholest-7-en-6-One是由上述结果假定的，并与培养系统合成并喂食。喂养实验表明，将中间体代谢为20-羟基甲虫比胆固醇更好。7。现在，我们提出3Beta-Hydroxy-5beta-cholest-7-en-6-One作为ecdysone生物合成早期的关键中级。我们目前的研究集中在鉴定该中间体的前体。