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目的:对青蒿酸(Ⅰ)进行生物转化研究,以获得具有新颖结构的青蒿酸衍生物。方法:将青蒿酸投入到对数生长期的三尖杉悬浮培养细胞和黄花蒿悬浮培养细胞中,共培养两天后终止转化,利用TLC及HPLC法对转化产物进行检测,利用硅胶柱,SephadexLH20以及ODS柱层析法分离纯化,利用波谱方法鉴定结构,最后利用HPLC考察共培养时间对转化率的影响。结果:青蒿酸在三尖杉与黄花蒿培养细胞中均发生了生物转化反应,分离鉴定出一个转化产物:3-α-羟基青蒿酸(Ⅱ)。三尖杉悬浮细胞和黄花蒿悬浮细胞转化Ⅱ的最佳共培养时间分别为2d与3d,总的摩尔转化率最高分别为8.42%与3.95%。结论:首次利用三尖杉悬浮细胞和黄花蒿悬浮细胞生物合成3-α-羟基青蒿酸。
OBJECTIVE: To study the biotransformation of artemisinic acid (Ⅰ) to obtain artemisinic acid derivatives with novel structure. METHODS: Artemisinic acid was injected into suspension culture cells of Cephalotaxus septentrionalis and Artemisia halodendron during logarithmic growth phase. After two days co-culture, the transformation was terminated. The products were detected by TLC and HPLC. The products were purified by silica gel column, Sephadex LH20 And ODS column chromatography separation and identification, the use of spectral methods to identify the structure, and finally the use of HPLC co-culture time on the conversion rate. Results: Artemisinic acid biotransformation occurred in cultured cells of Cephalotaxus and Artemisia annua. A transformed product was isolated and identified as 3-α-hydroxy artemisinic acid (Ⅱ). The best cocultivation time of the transformation of suspension cells of Cephalotaxus cuspidatus and A. annua was 2d and 3d, respectively, and the highest total molar conversion rates were 8.42% and 3.95% respectively. Conclusion: The first use of Cephalotaxus suspension cells and A. annua suspension cells biosynthesis of 3-α-hydroxy artemocarnic acid.