Santoyo-Garcia Jorge H, Valdivia-Cabrera Marissa, Ochoa-Villarreal Marisol, Casasola-Zamora Samuel, Ripoll Magdalena, Escrich Ainoa, Moyano Elisabeth, Betancor Lorena, Halliday Karen J, Loake Gary J, Rios-Solis Leonardo
Institute for Bioengineering, School of Engineering, University of Edinburgh, King's Buildings, Edinburgh, EH9 3FB, UK.
Centre for Engineering Biology, University of Edinburgh, King's Buildings, Edinburgh, EH9 3BF, UK.
Bioresour Bioprocess. 2023 Sep 29;10(1):68. doi: 10.1186/s40643-023-00687-8.
In this study, several approaches were tested to optimise the production and recovery of the widely used anticancer drug Taxol (paclitaxel) from culturable vascular stem cells (VSCs) of Taxus baccata, which is currently used as a successful cell line for paclitaxel production. An in situ product recovery (ISPR) technique was employed, which involved combining three commercial macro-porous resin beads (HP-20, XAD7HP and HP-2MG) with batch and semi-continuous cultivations of the T. baccata VSCs after adding methyl jasmonate (Me-JA) as an elicitor. The optimal resin combination resulted in 234 ± 23 mg of paclitaxel per kg of fresh-weight cells, indicating a 13-fold improved yield compared to the control (with no resins) in batch cultivation. This resin treatment was further studied to evaluate the resins' removal capacity of reactive oxygen species (ROS), which can cause poor cell growth or reduce product synthesis. It was observed that the ISPR cultivations had fourfold less intracellular ROS concentration than that of the control; thus, a reduced ROS concentration established by the resin contributed to increased paclitaxel yield, contrary to previous studies. These paclitaxel yields are the highest reported to date using VSCs, and this scalable production method could be applied for a diverse range of similar compounds utilising plant cell culture.
在本研究中,测试了几种方法以优化从欧洲红豆杉可培养的维管束干细胞(VSCs)中生产和回收广泛使用的抗癌药物紫杉醇的过程,欧洲红豆杉目前是用于紫杉醇生产的一种成功细胞系。采用了原位产物回收(ISPR)技术,该技术包括在添加茉莉酸甲酯(Me-JA)作为诱导剂后,将三种商业大孔树脂珠(HP-20、XAD7HP和HP-2MG)与欧洲红豆杉VSCs的分批和半连续培养相结合。最佳树脂组合在分批培养中每千克鲜重细胞可产生234±23毫克紫杉醇,表明与对照(不使用树脂)相比产量提高了13倍。进一步研究了这种树脂处理方法,以评估树脂对活性氧(ROS)的去除能力,活性氧会导致细胞生长不良或降低产物合成。观察到ISPR培养中的细胞内ROS浓度比对照低四倍;因此,树脂降低的ROS浓度有助于提高紫杉醇产量,这与之前的研究结果相反。这些紫杉醇产量是迄今为止使用VSCs报道的最高产量,这种可扩展的生产方法可应用于利用植物细胞培养生产各种类似化合物。