Pharmacognosy Department, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt.
Enzymology and Fungal Biotechnology Lab, Botany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig, 44519, Egypt.
Microb Cell Fact. 2024 Aug 16;23(1):229. doi: 10.1186/s12934-024-02495-x.
Epothilones are one of the common prescribed anticancer drugs for solid tumors, for their exceptional binding affinity with β-tubulin microtubule, stabilizing their disassembly, causing an ultimate arrest to the cellular growth. Epothilones were initially isolated from Sornagium cellulosum, however, their extremely slow growth rate and low yield of epothilone is the challenge. So, screening for a novel fungal endophyte dwelling medicinal plants, with higher epothilone productivity and feasibility of growth manipulation was the objective. Aspergillus niger EFBL-SR OR342867, an endophyte of Latania loddegesii, has been recognized as the heady epothilone producer (140.2 μg/L). The chemical structural identity of the TLC-purified putative sample of A. niger was resolved from the HPLC, FTIR and LC-ESI-MS/MS analyses, with an identical molecular structure of the authentic epothilone B. The purified A. niger epothilone B showed a resilient activity against MCF-7 (0.022 μM), HepG-2 (0.037 μM), and HCT-116 (0.12 μM), with selectivity indices 21.8, 12.9 and 4, respectively. The purified epothilone B exhibited a potential anti-wound healing activity to HepG-2 and MCF-7 cells by ~ 54.07 and 60.0%, respectively, after 24 h, compared to the untreated cells. The purified epothilone has a significant antiproliferative effect by arresting the cellular growth of MCF-7 at G2/M phase by ~ 2.1 folds, inducing the total apoptosis by ~ 12.2 folds, normalized to the control cells. The epothilone B productivity by A. niger was optimized by the response surface methodology, with ~ 1.4 fold increments (266.9 μg/L), over the control. The epothilone productivity by A. niger was reduced by ~ 2.4 folds by 6 months storage as a slope culture at 4 °C, however, the epothilone productivity was slightly restored with ethylacetate extracts of L. loddegesii, confirming the plant-derived chemical signals that partially triggers the biosynthetic genes of A. niger epothilones. So, this is the first report emphasizing the metabolic potency of A. niger, an endophyte of L. loddegesii, to produce epothilone B, that could be a new platform for industrial production of this drug.
埃坡霉素是一种常见的治疗实体瘤的抗癌药物，因其与β-微管蛋白的特殊结合亲和力，能够稳定微管蛋白的解聚，从而最终阻止细胞生长。埃坡霉素最初是从 Sornagium cellulosum 中分离出来的，然而，其极低的生长速度和低产量是一个挑战。因此，筛选出一种新型真菌内生植物，具有更高的埃坡霉素生产力和生长操纵的可行性，是我们的目标。黑曲霉 EFBL-SR OR342867 是一种 Latania loddegesii 的内生真菌，已被证实为埃坡霉素的高产菌（140.2μg/L）。通过 TLC 纯化的假定样品的化学结构鉴定通过 HPLC、FTIR 和 LC-ESI-MS/MS 分析得以解决，其分子结构与真正的埃坡霉素 B 相同。纯化的黑曲霉埃坡霉素 B 对 MCF-7（0.022μM）、HepG-2（0.037μM）和 HCT-116（0.12μM）具有较强的活性，其选择性指数分别为 21.8、12.9 和 4。与未经处理的细胞相比，纯化的埃坡霉素 B 在 24 小时后对 HepG-2 和 MCF-7 细胞的伤口愈合活性分别抑制了约 54.07%和 60.0%。与对照细胞相比，纯化的埃坡霉素 B 通过将 MCF-7 细胞的细胞生长阻滞在 G2/M 期，约增加 2.1 倍，通过总凋亡诱导，约增加 12.2 倍，从而显著抑制 MCF-7 细胞的增殖。黑曲霉的埃坡霉素 B 产量通过响应面法进行了优化，与对照相比，产量增加了约 1.4 倍（266.9μg/L）。然而，在 4°C 下作为斜板培养物储存 6 个月后，黑曲霉的埃坡霉素 B 产量降低了约 2.4 倍，而用拉坦尼 Loddegesii 的乙酸乙酯提取物处理后，埃坡霉素 B 的产量略有恢复，这证实了植物衍生的化学信号部分触发了黑曲霉埃坡霉素生物合成基因。因此，这是第一个强调黑曲霉（Latania loddegesii 的一种内生真菌）生产埃坡霉素 B 的代谢潜力的报告，这可能为该药物的工业生产提供一个新的平台。
