Aim To modify the structure of resibufogenin by using Ginkgo biloba suspension. Methods Young leaves of Ginkgo biloba were dedifferentiated into callus in MS medium with only 2,4-D as plant growth regulator. The callus was then transferred aseptically to liquid MS medium exogenously supplemented with appropriate concentration of 6-BA, NAA and 2,4-D to establish suspension cell culture system. Resibufogenin was administered into the well-grown cell cultures and incubated for 4 d. The products dissolved in the liquid phase of the cultures were extracted and purified by silica gel column chromatography gradiently eluted with petroleum ether and acetone system. Results One transformed product was obtained in 40% yield after 4 d incubation, which was identified as 3-epi-resibufogenin on the basis of FAB MS, 1H NMR and 13C NMR spectroscopic analysis and corresponding data reported in literature. Conclusion G. biloba suspension cultures can be used as an enzyme system to biotransform resibufogenin, an animal-originated bufadienolide, into 3-epi-resibufogenin.%目的利用银杏悬浮细胞对酯蟾毒配基进行结构修饰.方法利用只含生长素2,4-D的MS培养基诱导银杏嫩叶,使细胞脱分化形成愈伤组织,然后将愈伤组织转移至含一定浓度6-BA, NAA, 和2,4-D的液体MS培养基中以形成悬浮细胞.把酯蟾毒配基加入生长状态良好的悬浮细胞中转化四天.提取出溶解于液体相的转化产物, 采用硅胶吸附柱层析法, 以石油醚和丙酮为展开体系进行梯度洗脱, 然后对转化产物进行分离纯化.结果经过四天转化, 得到一个转化产物, 转化率达40%, 通过对转化产物的质谱, 核磁共振氢谱和碳谱等波谱数据进行分析, 并与有关文献进行对比, 证明转化产物为3-表-酯蟾毒配基.结论以银杏悬浮细胞作为一种生物酶体系,可以把来源于动物的蟾蜍甾烯类化合物酯蟾毒配基转化成3-表-酯蟾毒配基.
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