Sustainable use of Taxus media cell cultures through minimal growth conservation and manipulation of genome methylation
Sustainable use of Taxus media cell cultures through minimal growth conservation and manipulation of genome methylation
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通过最小限度的生长保护和基因组甲基化操作可持续利用红豆杉细胞培养物
DOI：
10.1016/j.procbio.2013.01.013
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发表时间：
2013-03-01
影响因子：
4.4
通讯作者：
Yu, Long-jiang
中科院分区：
文献类型：
--
作者：
Li, Li-qin;Li, Xia-li;Yu, Long-jiang
紫杉醇的产量随着曼地亚红豆杉细胞的重复传代而降低。我们使用最低限度的生长保护和基因组甲基化操作来维持曼地亚红豆杉细胞培养物的紫杉醇生产。低温（5 ℃）培养可使红豆杉细胞的继代周期延长至180 d。在保存后和第一次恢复传代培养周期中，细胞中的紫杉醇水平增加，然后在随后的恢复传代培养周期中下降。使用扩增片段长度多态性（AFLP）技术在这些文化的遗传变异分析确定只有两个多态性带与第二和第六个恢复周期的文化。然而，高效液相色谱的结果表明，DNA甲基化在重复传代过程中增加。用5-氮杂-2 '-脱氧胞苷处理引起的DNA甲基化水平的降低与紫杉醇水平的增加一致。同时暴露于茉莉酸甲酯和5-氮杂-2 '-脱氧胞苷使紫杉醇水平增加到320.43 μ g g（-1）（干重），这是保存前紫杉醇含量的六倍多。据我们所知，这是第一份关于通过确保红豆杉细胞的可持续利用来提高紫杉醇产量的报告。(C)2013爱思唯尔有限公司保留所有权利。
Yields of paclitaxel decreased with repeated subculturing of Taxus media cells. We used minimal growth conservation and manipulation of genome methylation to sustain paclitaxel production by Taxus media cell cultures. The subculture period of Taxus cells can be prolonged to 180 d by incubating them at a low temperature (5 degrees C). Paclitaxel levels increased in the cells after conservation and during the first recovery subculture cycle, and then decreased during the subsequent recovery subculture cycle. Analysis of genetic variations in these cultures using amplified fragment-length polymorphism (AFLP) technology identified only two polymorphic bands associated with the second and sixth recovery cycle cultures. However, the results of high-performance liquid chromatography indicated that DNA methylation increased during the course of repeated subculturing. A decrease in DNA methylation level caused by treatment with 5-Aza-2'-deoxycytidine coincided with an increase in paclitaxel levels. Simultaneous exposure to both methyl jasmonate and 5-Aza-2'-deoxycytidine increased paclitaxel levels to 320.43 mu g g(-1) (dry weight), which is more than six times the paclitaxel content before conservation. To our knowledge, this is the first report about improving paclitaxel production by ensuring sustainable use of Taxus cells. (C) 2013 Elsevier Ltd. All rights reserved.