刺五加内生放线菌Streptomyces sp. CWJ-256次生代谢产物研究  

Study on the secondary metabolites of an endophytic Streptomyces sp. CWJ-256 isolated from Acanthopanax senticosus

作　　者：王新位 郭文强 赵建元[1] 余利岩[1] 席楠[1] 何宁[1] 王珊珊 袁丽杰 解云英[1] WANG Xin-wei;GUO Wen-qiang;ZHAO Jian-yuan;YU Li-yan;XI Nan;HE Ning;WANG Shan-shan;YUAN Li-jie;XIE Yun-ying(Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences ＆ Peking Union Medical College,Beijing 100050,China;Hebei Key Laboratory for Chronic Diseases,Tangshan Key Laboratory for Preclinical and Basic Research on Chronic Diseases,School of Basic Medical Sciences,North China University of Science and Technology,Tangshan 063000,China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所,北京100050 [2]华北理工大学基础医学院/河北省慢性疾病重点实验室/唐山市慢性病临床基础研究重点实验室,唐山063000

出　　处：《中国医药生物技术》2018年第5期404-411,共8页Chinese Medicinal Biotechnology

基　　金：北京市自然科学基金(7172137);中国医学科学院医学与健康科技创新工程(2016-I2M-2-002);河北省自然科学基金(C2014209137)

摘　　要：目的研究刺五加植物内生放线菌CWJ-256次级代谢产物及其生物活性。方法采用16S rRNA基因同源进化分析对菌株CWJ-256进行初步分类鉴定,采用抗菌活性追踪方式,对CWJ-256发酵液通过硅胶柱层析、凝胶柱层析、薄层色谱显色以及高效液相色谱等方法对目标化合物分离纯化,利用核磁共振波谱法和质谱法鉴定化合物的结构。采用MTT法测试其抗肿瘤细胞增殖活性。基于Gluc报告系统对化合物3的体外抗流感病毒药效学进行初步评价。结果菌种鉴定显示Streptomycessp.CWJ-256为生黑孢链霉菌Streptomyces melanosporofaciens;从该菌株的发酵代谢产物中分离得到3个活性化合物,经结构鉴定为efomycin G(化合物1)、11,11'-O,O-dimethylelaiophylin(化合物2)、elaiophylin(化合物3);抗肿瘤细胞增殖活性评价显示,化合物1和3对体外培养的人乳腺癌细胞MDA-MB-231具有增殖抑制作用(IC50分别为4.385和2.118μmol/L)。体外抗流感病毒药效学评价显示,化合物3对流感病毒A/WSN/33(H1N1)具有一定抑制效果。结论刺五加内生放线菌Streptomycessp.CWJ-256可产生多个洋橄榄叶素类次生代谢产物,显示出抗细胞增殖、抗病毒等多种活性。Objective The aim of the study is to isolate and identify the bioactive secondary metabolites from the fermentation broth of the endophytic Streptomyces sp. CWJ-256 isolated from the plant Acanthopanax senticosus.Methods Phylogenetic analyses were based on 16S rRNA gene sequence. The bioactive metabolites were isolated by silica gel chromatography and preparative HPLC by guidance of antibacterial activity. HRMS and NMR were employed to characterize the structures of purified compounds. The anti-proliferation activities against human breast cancer cells were assayed by MTT, and the anti-influenza virus activities were tested by Gluc report system.Result Streptomyces sp. CWJ-256 was identified as Streptomyces melanosporofaciens. Three compounds were isolated and identified as efomycin G （1）, 11,11＇-O,O-dimethylelaiophylin （2）, elaiophylin （3）, respectively. Compounds 1 and 3 displayed anti-proliferation potential against human breast cancer cell MDA-MB-231 in vitro, and the compound 3 showed weak activity against influenza virus A/WSN/33（H1N1）.Conclusion The Streptomyces sp. CWJ-256 can produce elaiophylin and its analogues which exhibited antibacterial, antitumor and anti-virus activities.

关 键 词：刺五加△ 内生放线菌 洋橄榄叶素 抗肿瘤活性 抗流感病毒活性 

分 类 号：R915[医药卫生—微生物与生化药学]