Division of Biotechnology, Institute of Himalayan Bioresource Technology, CSIR, Palampur176 061 H.P., India.
Cell Biol Int. 2011 Feb;35(2):153-8. doi: 10.1042/CBI20090459.
Cell suspension cultures of Arnebia euchroma were raised from in vitro leaf-derived friable callus on liquid MS [Murashige and Skoog] medium supplemented with BAP (6-benzylaminopurine) (10.0 μM) and IBA (indole-3-butyric acid) (5.0 μM). A two-stage culture system was employed using growth and production medium for cell biomass and shikonin derivatives, respectively. Factors such as light, temperature, sucrose and pH (hydrogen ion concentration) were studied to observe their effect on the shikonin derivative production. Light conditions completely inhibited shikonin derivative production. Out of different temperature regimes tested, the highest yield (586.17 μg/g FW) was found at 25°C. Maximum production (656.14 μg/g FW) was observed in 6% sucrose. An alkaline pH (7.25-9.50) favoured shikonin derivative production. The results showed that physical and chemical factors greatly influence the production of shikonin derivatives in cell suspension cultures of A. euchroma. Therefore, by employing optimum culture conditions, it is possible to enhance the production of secondary compounds from the cells. The factors optimized for in vitro production of shikonin derivatives during the present study can successfully be employed for their large-scale production in bioreactors.
从离体叶衍生的易碎愈伤组织在液体 MS [Murashige 和 Skoog]培养基上培养出了Arnebia euchroma 的细胞悬浮培养物，该培养基添加了 BAP（6-苄基氨基嘌呤）（10.0 μM）和 IBA（吲哚-3-丁酸）（5.0 μM）。使用生长培养基和生产培养基进行了两阶段培养系统，分别用于细胞生物量和紫草素衍生物的生产。研究了光照、温度、蔗糖和 pH（氢离子浓度）等因素，以观察它们对紫草素衍生物生产的影响。光照条件完全抑制了紫草素衍生物的生产。在测试的不同温度条件下，在 25°C 时产量最高（586.17 μg/g FW）。在 6%的蔗糖中观察到最大产量（656.14 μg/g FW）。碱性 pH（7.25-9.50）有利于紫草素衍生物的生产。结果表明，物理和化学因素极大地影响了 A. euchroma 细胞悬浮培养物中紫草素衍生物的生产。因此，通过采用最佳培养条件，可以提高细胞中次生化合物的产量。本研究中优化的用于紫草素衍生物体外生产的因素可以成功地用于生物反应器中的大规模生产。
