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Taxol promising fungal endophyte, Pestalotiopsis species isolated from Taxus cuspidata

机译:紫杉醇有望真菌内生菌,从东北红豆杉分离出的Pestalotiopsis种

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摘要

The endophytic fungi, Pestalotiopsis versicolor and Pestalotiopsis neglecta, were isolated from the healthy leaves and bark of the Japanese Yew tree, Taxus cuspidata. The fungal species were identified by their characteristic culture morphology and molecular analysis. For the first time, the test fungi were screened for the production of taxol in modified liquid medium The presence of taxol was confirmed by HPLC, ~1H NMR, and LC-MS methods of analysis. The maximum amount of taxol production in P. versicolor was recorded as 478 μg/l. The production rate was increased to 9560-fold than that found in the culture broth of earlier reported fungus, Taxomyces andreanae. The extracted fungal taxol showed a strong cytotoxic activity in the in vitro culture of tested human cancer cells by apoptotic assay indicating that the increase in taxol concentration induces increased cell death. A PCR-based screening for ts, a unique gene in the formation of the taxane skeleton, confirmed the molecular blueprint for taxol biosynthesis. The results designate that the fungal endophyte, P. versicolor, is an excellent candidate for an alternate source of taxol supply and can also serve as a potential species for genetic engineering to enhance the production of taxol to a higher level.
机译:从紫杉紫杉(Taxus cuspidata)的健康叶子和树皮中分离出内生真菌,即杂种天蛾(Pestalotiopsis versicolor)和疏生天蛾Pestalotiopsis neglecta。通过其特征性的培养形态和分子分析来鉴定真菌种类。首次筛选了测试真菌在改良液体培养基中生产紫杉醇的能力。紫杉醇的存在通过HPLC,〜1H NMR和LC-MS分析方法进行了确认。杂色青霉中紫杉醇的最大产量记录为478μg/ l。与早期报道的真菌Taxomyces andreanae的培养液相比,生产率提高了9560倍。提取的真菌紫杉酚通过凋亡分析在被测人癌细胞的体外培养中显示出强大的细胞毒性活性,表明紫杉酚浓度的增加诱导细胞死亡的增加。 ts(紫杉烷骨架形成过程中的独特基因)基于PCR的筛选证实了紫杉醇生物合成的分子蓝图。结果表明,真菌内生菌杂色种(P. versicolor)是紫杉醇供应的替代来源的极佳候选者,并且还可以作为基因工程的潜在物种,以将紫杉醇的产量提高到更高水平。

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