摘要
目的从南方红豆杉(Taxus chinensis var.mairei)中分离产紫杉醇内生真菌,并研究其抗肿瘤细胞活性。方法采用组织块法从南方红豆杉的树根部、树主干内层、侧枝及针叶分离纯化内生真菌,经液体发酵培养,抽提发酵粗产物,以标准紫杉醇为对照进行薄层层析(TLC)、紫外光谱(UV)、HPLC、质谱(MS)分析。同时将经上述方法确认后的内生真菌产的紫杉醇作用于人宫颈癌HeLa细胞,采用MTT法分析其抗肿瘤细胞活性。结果从南方红豆杉主干韧皮部分离得到1株产紫杉醇内生真菌D62,其紫杉醇产量为148.95μg/L。对菌株D62的菌落和菌丝体的形态学分析表明其属于镰刀属(Fusarium sp.)。将该菌株产紫杉醇作用于HeLa细胞,表现出明显的致细胞凋亡作用。结论南方红豆杉中分离得到的产紫杉醇内生真菌D62具有优良的发酵特性。
Objective To isolated from producing endophytic fungi of Taxus chinensis var.mairei,and study its anti-tumor activity.
Methods The hyphal tips,which grew out from the tissue of Taxus chinensis var.mairei were isolated and transferred to potato dextrose agar （PDA）.Then the purified endophytic fungal strains were fermented.The presence of taxol in the fungal extract was confirmed by thin-layer chromatography （TLC）,ultraviolet spectrophotometry （UV）,high performance liquid chromatography （HPLC） and mass spectrometry （MS） by comparison with taxol standard.The anti-tumor activity of the fungal taxol was detected by Methyl Thiazolyl Tetrazolium （MTT） method with HeLa cells.
Results Strain D62,an isolated endophytic fungus from the phloem of the Taxus chinensis var.mairei,was observed to produce taxol,when grown in semi-synthetic liquid media.The amount of taxol produced by this endophytic fungus was 148.95 μg/L determined by HPLC.The fungus was identified as a Fusarium sp.according to the morphology of the fungal culture and the characteristics of the colony.The fungal taxol extracted also showed a strong cytotoxic activity in the in vitro culture of HeLa cells tested by an apoptotic assay.
Conclusion Strain D62,a taxol-producing endophytic fungus isolated from Taxus chinensis var.mairei,can serve as a potential material for fungal fermentation to improve taxol production.
出处
《中国医药生物技术》
CSCD
2010年第3期202-207,共6页
Chinese Medicinal Biotechnology
关键词
抗肿瘤药
植物
真菌
发酵
红豆杉属
Antineoplasstic agents
phytogenic
Fungi
Fermentation
Taxol
