【作者】 胡艳山；

【导师】 于荣敏；

【作者基本信息】 暨南大学 ， 药物化学， 2010， 硕士

【摘要】 目的：利用三尖杉和转基因西洋参冠瘿组织培养体系对青蒿素的生物合成中间体青蒿酸及二氢青蒿酸进行生物转化研究,获得三个青蒿酸及二氢青蒿酸衍生物,同时得到一个次级代谢产物。方法：通过TLC和HPLC方法检测转化体系中的转化产物；利用硅胶柱层析、制备薄层、Sephadex LH20、ODS柱等方法分离纯化转化产物；检验产物的理化性质；用1H-NMR,13C-NMR和MS方法鉴定转化产物结构；用HPLC方法考察转化体系对底物进行生物转化的最佳共培养条件。结果：国内首次使用三尖杉悬浮培养细胞进行生物转化研究,以青蒿素的生物合成途径中两个重要中间体青蒿酸和二氢青蒿酸为底物。青蒿酸作为底物与三尖杉悬浮培养细胞共培养两天,从三尖杉体系分离并鉴定一个产物3α-羟基青蒿酸(SQJ-2),最佳共培养时间为2d,总的摩尔转化率最高为8.42%。共培养时间也为两天,从体系中分离并鉴定两个转化产物分别为：15-羟基二氢青蒿酸(SQQJ-3)、15-羟基二氢青蒿酸-β-D-葡萄糖酯(SQQJ-5);培养体系中SQQJ-3的转化率在共培养的第2d达到最大值(6.02%),SQQJ-5在共培养的第3d达到最大值(4.95%)。使用转基因西洋参冠瘿组织对青蒿酸进行转化得到一个次级代谢产物：12,13-环氧-11-羟基-9-十八碳烯酸。更多还原

【Abstract】 Abstract Objective:To investigate the biotransformation of artemisinic acid and dihydroartemisinic acid by cell suspension culture of Cephalotaxus fortunei Hook. f. and transgenic crown gall of Panax quinquefolium L, generated three transformed Products and a secondary metabolite.Methods:Plant tissue culture technology was employed. Transformed products were detected by TLC and HPLC, isolated and purified by the column chromatography, recrystallization, Sephadex LH20, ODS etc. Their physico-chemical properties were checked up. Their structures were elucidated by 1H-NMR, 13C-NMR and MS. The optimal co-culture times of the products were determined by HPLC.Results:The cell suspension culture of Cephalotaxus fortunei Hook. f. transformed the artemisinic acid and dihydroartemisinic acid to three Products, transformed artemisinic acid to one Product:3a-hydroxyartemisinic acid. After co-cultured for 2 days, the mole conversion ratio of artemisinic acid reached the highest (8.42%) in cell suspension culture of Cephalotaxus fortunei Hook. f. The cell suspension culture of Cephalotaxus fortunei Hook. f. transformed dihydroartemisinic acid to two Products: 15-hydroxy dihydroartemisinic acid and 15-hydroxy dihydroartemisinic acidβ-D-glucopyranosyl ester. The conversion rate of SQQJ-3 in the cultures reached the maximum (6.02%) in 2nd day, and conversion rate of SQQJ-5 in the cultures reached the maximum (4.95%) in 3rd day. The transgenic crown gall of Panax quinquefolium L transformed artemisinic acid, generated a secondary metabolite:12,13-epoxy-11-hydroxy-9-octadecenoic acid.更多还原