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在东北红豆杉悬浮培养物中提高紫杉醇和浆果赤霉素III的产量。

Improved paclitaxel and baccatin III production in suspension cultures of Taxus media.

作者信息

Cusidó Rosa M, Palazón Javier, Bonfill Mercedes, Navia-Osorio Alberto, Morales Carmen, Piñol M Teresa

机构信息

Laboratorio de Fisiologia Vegetal, Facultad de Farmacia, Universidad de Barcelona, Avda. Diagonal 643, 08028 Barcelona, Spain.

出版信息

Biotechnol Prog. 2002 May-Jun;18(3):418-23. doi: 10.1021/bp0101583.

Abstract

A cell suspension culture of Taxus media was established from a stable callus line of this species. The growth rate and production of paclitaxel and baccatin III of this cell suspension were significantly increased during the shake flask culture in its respective optimum media for cell growth and product formation, which were selected after assaying 24 different culture media. The highest yields of paclitaxel (2.09 mg L(-1)) and baccatin III (2.56 mg L(-1)) in the production medium rose (factors of 7.0 and 3.0, respectively) in the presence of methyljasmonate (220 microg g(-1) FW). When the elicitor was added together with mevalonate (0.38 mM) and N-benzoylglycine (0.2 mM), the increase in the yields of paclitaxel and baccatin III was even higher (factors of 8.3 and 4.0, respectively). Thereafter, a two-stage culture for cell suspension was carried out using a 5-l stirred bioreactor running for 36 days, the first stage being in the cell growth medium until cells entered their stationary growth phase (12 days) and the second stage being in the production medium supplemented with the elicitor and two putative precursors in the concentrations indicated above. Under these conditions, 21.12 mg L(-1) of paclitaxel and 56.03 mg L(-1) of baccatin III were obtained after 8 days of culture in the production medium.

摘要

从曼地亚红豆杉的一个稳定愈伤组织系建立了其细胞悬浮培养物。在24种不同培养基经过测定后,选择了该细胞悬浮培养物在其各自用于细胞生长和产物形成的最佳培养基中进行摇瓶培养时,其生长速率以及紫杉醇和巴卡亭III的产量显著增加。在存在茉莉酸甲酯(220μg g(-1) FW)的情况下,生产培养基中紫杉醇(2.09 mg L(-1))和巴卡亭III(2.56 mg L(-1))的最高产量有所提高(分别提高了7.0倍和3.0倍)。当诱导剂与甲羟戊酸(0.38 mM)和N-苯甲酰甘氨酸(0.2 mM)一起添加时,紫杉醇和巴卡亭III产量的增加甚至更高(分别提高了8.3倍和4.0倍)。此后,使用一个5升搅拌式生物反应器进行了为期36天的细胞悬浮两阶段培养,第一阶段在细胞生长培养基中进行,直到细胞进入稳定生长期(12天),第二阶段在添加了上述浓度的诱导剂和两种假定前体的生产培养基中进行。在这些条件下,在生产培养基中培养8天后,获得了21.12 mg L(-1)的紫杉醇和56.03 mg L(-1)的巴卡亭III。

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