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目的利用转基因何首乌Polygonum multiflorum毛状根对青蒿酸进行生物转化研究,分离鉴定其转化产物。方法何首乌毛状根预培养7 d,投入青蒿酸,培养2 d,终止反应,利用TLC和GC-MS对转化产物进行检测,利用硅胶柱、ODS反相柱和Sephadex LH-20柱色谱对转化产物进行分离纯化,并根据理化数据和波谱技术鉴定转化产物的化学结构。结果青蒿酸在何首乌毛状根中发生了转化反应,经GC-MS检测可生成多种青蒿素类化合物。分离鉴定了2个转化产物:异青蒿内酯(1)和3β-羟基青蒿酸(2),利用GC-MS鉴定了另外2个转化产物:去氧青蒿素B(3)和青蒿内酯(4)。结论本实验首次利用转基因植物器官对青蒿酸进行生物转化研究,得到3个青蒿素类化合物和1个羟基化产物。该研究一方面填补了转基因植物器官对青蒿素类化合物生物转化的空白,另一方面也丰富了转基因何首乌毛状根的化合物转化类型。
OBJECTIVE: To study the biotransformation of artemisinic acid with Polygonum multiflorum hairy root, and to isolate and identify the transformed product. Methods The hairy roots of Polygonum multiflorum were pre-cultured for 7 days. Artemisinic acid was added into the solution for 2 days to terminate the reaction. The products were detected by TLC and GC-MS. Silica gel column, ODS reverse phase column and Sephadex LH-20 column chromatography The products were separated and purified, and the chemical structures of the products were identified by physicochemical data and spectroscopic techniques. Results Artemisinic acid was transformed in the hairy root of Polygonum multiflorum. A variety of artemisinin compounds were produced by GC-MS. Two transformation products were identified: iso-arteralactone (1) and 3β-hydroxyartemisinic acid (2), and the other two transformation products were identified by GC-MS: deoxyartemisinin B (3) Arthanolactone (4). CONCLUSIONS In this experiment, we first used the organs of transgenic plants to study the biotransformation of artemisinic acid. Three artemisinin compounds and one hydroxylation product were obtained. This research, on the one hand, fills the blank of the biotransformation of artemisinin compounds in organs of transgenic plants and, on the other hand, enriches the types of compound transformation of hairy roots of transgenic Polygonum multiflorum.