一株产紫杉醇菌的分离及其代谢产物对HeLa细胞抑制作用的研究

Isolation of a taxol-producing endophytic fungus and inhibiting effect of the fungus metabolites on HeLa cell

从东北红豆杉Taxus cuspidata的主干和侧枝树皮及皮下分离筛选得到181株纯化的内生菌株。经过摇瓶发酵培养,再用甲醇、乙酸乙酯萃取抽提,抽提产物经以标准紫杉醇为对照的薄层层析(TLC)、高效液相色谱(HPLC)、质谱(MS)分析,发现有1株菌株HD181-23能够产生紫杉醇,其紫杉醇产量为206.34μg/L。对菌株HD181-23菌落、菌丝体和孢子形态和产孢特征进行了研究,表明菌株HD181-23为葡萄孢属的种,暂定为Botrytissp.。该菌株的发酵提取纯化产物经HeLa细胞毒实验,结果显示对HeLa细胞具有较强的抑制作用。

One hundred and eighty one purified endophytic fungal strains were isolated and screened from inner bark （phloem-cambium） and xylem of Taxus cuspidata. These strains were fermented and then the fermented solution was extracted using methanol and ethyl acetate. The extracts were analyzed by thin-layer chromatography （TLC）, high performance liquid chromatography （HPLC） and mass spectrum （MS）, using standard taxol as control. The results showed that strain HD181-23 can produce taxol with output of 206.34μg/L. Morphological observation indicates that HD181-23 belongs to the genus Botrytis, and the fungus is tentatively identified as Botrytis sp. The purified products of the fermented extracts have significant inhibitive effects on HeLa cell.