转基因何首乌毛状根生物转化青蒿酸的研究

Biotransformation of artemisinic acid by transgenic hairy roots of Polygonum multiflorum

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 investigate the biotransformation of artemisinc acid by transgenic hairy roots of Polygonum multiflorum and identify the transformation products.Methods Artemisinic acid was added to the suspension of transgenic hairy roots of P.multiflorum which had been pre-cultured for 7 d and co-cultured for another 2 d.The biotransformation products were detected by TLC and GC-MS,isolated by various chromatographic methods,and identified by physicochemical analyses and spectral data.Results GC-MS results showed that artemisinic acid could be biotransformed to many kinds of artemisinic compounds in the suspension of transgenic hairy roots of P.multiflorum.Two biotransformed products,isoannulide（1） and 3β-hydroxyartemisinic acid（2）,were isolated.In addition,deoxyarteannuin B（3） and annulide（4） were determined by GC-MS.Conclusion It is the first time to biotransform artemisinic acid by transgenic organs and to obtain three artemisinic compounds and one hydroxylation product,which fills the gaps in biotransformation of artemisinic compounds by transgenic organs and also enriches compound types of transgenic hairy roots of P.multiflorum.