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作 者:赵凯[1] 赵立斐[1] 金媛媛[1] 魏海霞[1] 平文祥[1] 周东坡[1]
机构地区:[1]黑龙江省高校微生物重点实验室黑龙江大学生命科学学院,哈尔滨150080
出 处:《菌物学报》2008年第5期735-744,共10页Mycosystema
基 金:国家自然科学基金项目(No.30570025);黑龙江省"十五"重大攻关项目(No.GA02C101);哈尔滨市青年科学基金项目(No.2005AFQXJ063);黑龙江省教育厅海外学人科研资助项目(No.1152HZ06);黑龙江大学杰出青年基金项目
摘 要: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.
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