巴山榧树内生真菌产生的二萜类物质的鉴别及对Hela细胞的抑制作用  

Identification of the diterpenoids produced by the endophytic fungi from Torreya fargesii and inhibitory effect on Hela cells

作　　者：金卫华 曹军卫[1] 涂毅[1] 唐艺 葛然 陈静文 徐甫 熊琦婕 JIN Wei-hua;CAO Jun-wei;TU Yi;TANG Yi;GE Ran;CHEN Jing-wen;XU Fu;XIONG Qi-jie(De-partment of Life Science and Chemistry,Wuhan Donghu University,Hubei Wuhan 430212,China)

机构地区：[1]武汉东湖学院生命科学与化学学院,湖北武汉430212

出　　处：《中国医院药学杂志》2018年第18期1898-1901,共4页Chinese Journal of Hospital Pharmacy

基　　金：湖北高校2017年省级大学生创新创业训练计划项目(编号:201711798030);湖北省普通高等学校"战略性新兴(支柱)产业人才培养计划"资助项目[湖北省教育厅鄂教高[2013]11号];省教育厅;省财政厅"专业综合改革"试点资助项目[鄂教高办[2014]6号];湖北省教育科学"十二五"规划项目(编号:2014B272);校青年自然科学基金(编号:2013dhzk003)

摘　　要：目的:研究巴山榧树组织分离产生二萜类物质紫杉醇的植物内生菌及其产生的紫杉醇类似物对肿瘤细胞的抑制作用。方法:分离纯化出植物内生菌,研究其对革兰阳性(G+)和革兰阴性(G-)细菌的抗性,并对菌株BP6T3进行了分类鉴定,用高效液相色谱(HPLC)分析其代谢产物的保留时间、百分含量,液质联用仪(HPLC-MS)分析该物质形成离子片段的特征。初步探讨紫杉醇物质对Hela细胞的抑制作用,并采用添加前体物质等,以提高代谢产物产量。结果:分离得到的菌株BP6T3产生的次生代谢产物具有抗G+和G-细菌的双抗活性,该菌株经初步分类鉴定为青霉sp(Penicilliumsp)。其发酵液提取物保留时间为12.8 min,与紫杉醇对照品的保留时间基本一致。百分含量为15.8%,发酵液中紫杉醇类似物的含量达到16.59mg·L-1。HPLC-MS测定其代谢抽提物中该物质形成离子片段的特征,表明其离子片段信息与紫杉醇对照品完全一致,可初步判定青霉BP6T3可产生紫杉醇类化合物。该菌株产生的紫杉醇物质对Hela细胞有明显的抑制作用,并随着浓度的增加而增加。不同前体物质对该菌株产生紫杉醇代谢物有明显的影响,以L-苯丙氨酸为前体,产量增加最多,增长率为425.17%。结论:菌株BP6T3可被用于大规模生产抗癌药物紫杉醇。OBJECTIVE To dissociate endophytic fungus producing diterpenoids taxol fromTorreyafargesii tissue and examine its inhibiting effect on tumor cells.METHODS Plant endophytes were isolated and purified to study their resistance to gram-positive（G＋）and gram-negative bacteria（G-）.High performance liquid chromatography（HPLC）was used for analysis of the retention time,relative peak area and percentage content of its metabolite.By liquid chromatography-mass spectrometry（HPLC-MS）,the material characteristics of the ion pair information of the metabolites were measured.The bacterial strain was also classifid.RESULTS The results showed that the secondary metabolites produced by the strain BP6 T3 possessed double resistance to G＋and G-bacteria.The strain was identified as Penicilliumsp by preliminary classification.Through HPLC analysis,the retention time of fermentation extracts was 12.8 min with almost the same as the standard of taxol.The average relative percentage was about 15.8%,and the content of taxol analogs in fermentation broth reached 16.59 mg·L-1.The material characteristic of the formation of ion fragments of taxane analogues in metabolic extracts was identical to that of the taxol standard determined by HPLC-MS.It could be initially determined that strain BP6 T3 could produce taxane compounds.Taxol substance produced by this strain had obvious inhibitory effects on Hela cells with the concentration increasing.Different precursors had significant effects on the production of paclitaxel metabolites in this strain.L-phenylalanine was used as the precursor and the yield increased most,with an increase rate of 425.7%.CONCLUSION The strain is expected to be used for mass production in antitumor drug taxol.

关 键 词：植物内生菌 紫杉醇 HELA细胞 抑制作用 

分 类 号：R93[医药卫生—生药学]