Department of Agricultural Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran.
Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, 1983969411 Tehran, Iran.
J Photochem Photobiol B. 2022 Sep;234:112532. doi: 10.1016/j.jphotobiol.2022.112532. Epub 2022 Jul 23.
Taxus baccata L. cell culture is a promising commercial method for the production of taxanes with anti-cancer activities. In the present study, a T. baccata cell suspension culture was exposed to white light and 2-aminoindan-2-phosphonic acid (AIP), a phenylalanine ammonia lyase (PAL) inhibitor, and the effects of this treatment on cell growth, PAL activity, total phenol content (TPC), total flavonoid content (TFC), taxane production and the expression of some key taxane biosynthetic genes (DXS, GGPPS, T13OH, BAPT, DBTNBT) as well as the PAL were studied. Light reduced cell growth, whereas AIP slightly improved it. Light increased PAL activity up to 2.7-fold relative to darkness. The highest TPC (24.89 mg GAE/g DW) and TFC (66.94 mg RUE/g DW) were observed in cultures treated with light and AIP. Light treatment also resulted in the maximum content of total taxanes (154.78 μg/g DW), increasing extracellular paclitaxel and cephalomannin (3.3-fold) and intracellular 10-deacetyl paclitaxel (2.5-fold). Light significantly increased the expression level of PAL, DBTNBT, BAPT, and T13αOH genes, whereas it had no effect on the expression of DXS, a gene active at the beginning of the taxane biosynthetic pathway. AIP had no significant effect on the expression of the target genes. In conclusion, the light-induced activation of PAL transcription and altered expression of relevant biosynthetic genes reduced cell growth and increased the content of total phenolic compounds and taxanes. These findings can be applied to improve taxane production in controlled cultures and bioreactors.
欧洲红豆杉细胞培养是生产具有抗癌活性的紫杉烷的有前途的商业方法。在本研究中,欧洲红豆杉细胞悬浮培养物暴露于白光和 2-氨基茚满-2-磷酸(AIP),一种苯丙氨酸解氨酶(PAL)抑制剂,并研究了这种处理对细胞生长、PAL 活性、总酚含量(TPC)、总类黄酮含量(TFC)、紫杉烷产量以及一些关键紫杉烷生物合成基因(DXS、GGPPS、T13OH、BAPT、DBTNBT)以及 PAL 的表达的影响。光照会降低细胞生长,而 AIP 则略微提高了细胞生长。与黑暗相比,光照使 PAL 活性增加了 2.7 倍。在光照和 AIP 处理的培养物中观察到最高的 TPC(24.89 mg GAE/g DW)和 TFC(66.94 mg RUE/g DW)。光照处理还导致总紫杉烷含量最高(154.78 μg/g DW),增加了细胞外紫杉醇和 Cephalomannin(3.3 倍)和细胞内 10-去乙酰紫杉醇(2.5 倍)。光照显著增加了 PAL、DBTNBT、BAPT 和 T13αOH 基因的表达水平,而对紫杉烷生物合成途径起始基因 DXS 的表达没有影响。AIP 对靶基因的表达没有显著影响。总之,PAL 转录的光诱导激活和相关生物合成基因的改变表达降低了细胞生长,增加了总酚类化合物和紫杉烷的含量。这些发现可应用于改善受控培养物和生物反应器中的紫杉烷产量。