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红豆杉(Taxus)细胞内紫杉醇的含量低且不稳定,限制了利用细胞培养大规模生产紫杉醇的产业化进程。以紫杉醇含量较高的曼地亚红豆杉(Taxus m edia)为试材,诱导得到了曼地亚红豆杉愈伤组织,在添加抗坏血酸(VC)、聚乙烯吡咯烷酮(PVP)、活性炭(activated carbon)的改良B5培养基上,15次反复继代培养后获得了高产紫杉醇的细胞系。对新建立的曼地亚红豆杉细胞系(MC)与同期诱导的东北红豆杉细胞系(NC)及实验室继代多年的中国红豆杉细胞系(SC)在细胞生长周期、紫杉醇含量和细胞死亡率等方面进行了分析比较。结果表明,SC的生长量达5.9倍,高于MC和NC的3.6和4.2倍。在初始接种量相近的情况下,MC的悬浮培养细胞在一个周期内紫杉醇产量可达9.5 mg/L,经过茉莉酸甲酯(M J)诱导后可达到41 mg/L。
The low and unstable taxol intracellular paclitaxel limits the industrialization of large-scale paclitaxel production using cell culture. Taxus mairei (Taxus m edia) with high paclitaxel content was used as experimental material to induce callus of Taxus mairei. After adding ascorbic acid (VC), polyvinylpyrrolidone (PVP), activated carbon ) On the modified B5 medium, 15 times repeatedly subculture obtained paclitaxel-producing cell lines. In the cell growth cycle of newly established Taxus mairei cell lines (MC) and concurrently induced Taxus cuspidata cell lines (NC) and laboratory subcultivated Taxus chinensis cell lines (SC) for many years, the contents of paclitaxel and cells Mortality and other aspects were analyzed and compared. The results showed that the growth of SC was 5.9 times higher than 3.6 and 4.2 times of that of MC and NC. Under similar initial inoculum size, paclitaxel production was up to 9.5 mg / L in one cycle and 41 mg / L after induction by methyl jasmonate (M J).