Kumar Pawan, Saini Mahak, Bhushan Shashi, Warghat Ashish R, Pal Tarun, Malhotra Nikhil, Sood Archit
Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Post Box No. 6, Palampur, 176061, Himachal Pradesh, India,
Appl Biochem Biotechnol. 2014 May;173(1):248-58. doi: 10.1007/s12010-014-0838-x. Epub 2014 Mar 19.
Cell suspension cultures of Arnebia euchroma were established from the friable callus on liquid Murashige and Skoog medium supplemented with 6-benzylaminopurine (10.0 μM) and indole-3-butyric acid (5.0 μM). Salicylic acid was used to study its effect on the enzymes which participate in shikonin biosynthesis with respect to metabolite (shikonin) content in the cell suspension culture of A. euchroma. In our study, phenylalanine ammonia lyase and PHB geranyltransferase were selected from the entire biosynthetic pathway. Results showed that phenylalanine ammonia lyase is responsible for growth and PHB geranyltransferase for metabolite production. Salicylic acid exhibited an inverse relationship with the metabolite content (shikonin); salicylic acid (100 μM) completely inhibited shikonin biosynthesis. The results presented in the current study can be successfully employed for the metabolic engineering of its biosynthetic pathway for the enhancement of shikonin, which will not only help in meeting its industrial demand but also lead to the conservation of species in its natural habitat.
从添加了6-苄基腺嘌呤（10.0 μM）和吲哚-3-丁酸（5.0 μM）的液体Murashige和Skoog培养基上的易碎愈伤组织中建立了新疆紫草的细胞悬浮培养物。使用水杨酸研究其对参与紫草素生物合成的酶的影响，该影响与新疆紫草细胞悬浮培养物中的代谢物（紫草素）含量有关。在我们的研究中，从整个生物合成途径中选择了苯丙氨酸解氨酶和紫草素香叶基转移酶。结果表明，苯丙氨酸解氨酶负责生长，而紫草素香叶基转移酶负责代谢物的产生。水杨酸与代谢物含量（紫草素）呈负相关；水杨酸（100 μM）完全抑制了紫草素的生物合成。本研究中呈现的结果可成功用于其生物合成途径的代谢工程，以提高紫草素的产量，这不仅有助于满足其工业需求，还将有助于保护其自然栖息地中的物种。
