Department of Biochemistry, Indian Institute of Science, Bangalore, 560012, India.
Biorefining and Advanced Materials Research Center, Scotland's Rural College (SRUC), SRUC Barony Campus, Parkgate, Dumfries, DG1 3NE, UK.
Sci Rep. 2020 Dec 3;10(1):21139. doi: 10.1038/s41598-020-77605-x.
Taxol (paclitaxel), a plant-derived anticancer drug, has been among the most successful anticancer drugs of natural origin. Endophytic fungi have been proposed as a prominent alternative source for Taxol and its intermediate Baccatin III, however the very low yields remain a hinderance to their commercial utilization. Significant research efforts towards this end are underway globally. Here, we report the results on our earlier reported Taxol-producing endophytic fungus, Fusarium solani from the standpoint of spores as seed inoculum and media selection for enhanced Taxol and baccatin III yields. Spores produced on M1D medium with 94.76% viability were used for further media optimization for Taxol and Baccatin III production in five different liquid media under static and shaker condition at different cultivation days. Taxol and Baccatin III when quantified through competitive inhibition enzyme immunoassay (CIEIA), showed maximum production at 136.3 µg L and 128.3 µg L, respectively in the modified flask basal broth (MFBB) under shaking condition. Further, two important genes of this pathway, namely taxane 13α-hydroxylase (T13αH) and 10-deacetylbaccatin III-10-β-O-acetyltransferase (DBAT) have been identified in this fungus. These findings are hoped to assist in further manipulation and metabolic engineering of the parent F. solani strain towards the enhanced production of Taxol and baccatin III.
紫杉醇（紫杉醇）是一种植物来源的抗癌药物，是最成功的天然来源抗癌药物之一。内共生真菌已被提议作为紫杉醇及其中间产物巴卡丁 III 的重要替代来源，然而，非常低的产量仍然是其商业利用的障碍。全球正在为此进行大量的研究工作。在这里，我们根据 earlier reported Taxol-producing endophytic fungus（先前报道的产紫杉醇内生真菌）、Fusarium solani（茄病镰刀菌）从孢子作为种子接种体和培养基选择的角度，报告了有关提高紫杉醇和巴卡丁 III 产量的结果。在 M1D 培养基上产生的具有 94.76%活力的孢子用于进一步优化五种不同液体培养基中的 Taxol 和 Baccatin III 生产，在不同培养天数下在静态和摇床条件下进行。通过竞争性抑制酶免疫分析（CIEIA）定量紫杉醇和巴卡丁 III 时，在摇床条件下改良瓶基础肉汤（MFBB）中分别显示出最高产量 136.3µg L 和 128.3µg L。此外，还在这种真菌中鉴定了该途径的两个重要基因，即紫杉醇 13α-羟化酶（T13αH）和 10-去乙酰基巴卡丁 III-10-β-O-乙酰基转移酶（DBAT）。这些发现有望有助于进一步对内共生真菌亲本 F. solani 菌株进行操作和代谢工程改造，以提高紫杉醇和巴卡丁 III 的产量。
