茯砖茶中冠突散囊菌的次级代谢产物及其生物活性研究

Secondary metabolites of Eurotium cristatum from Fu Brick Tea and their biological activities

目的研究茯砖茶中冠突散囊菌Eurotium cristatum的次级代谢产物及其生物活性。方法采用正、反相硅胶柱、凝胶柱和薄层制备等色谱方法和重结晶技术进行分离纯化,通过各种波谱技术进行结构鉴定;以3种肿瘤细胞SF-268、MCF-7和NCI-H460为供试细胞株,采用SRB法对化合物进行细胞毒活性研究;以金黄色葡萄球菌、大肠杆菌、肠炎沙门氏菌肠炎亚种、痢疾志贺氏菌和普通变形杆菌等肠道致病菌为供试细菌,采用MTT法对化合物进行抗菌活性研究。结果从茯砖茶中冠突散囊菌发酵液提取物中分离得到9个化合物,分别鉴定为大黄素甲醚(1)、1,5-二羟基-3-甲氧基-7-甲基蒽醌(2)、灰绿曲霉黄色素(3)、2-(2′,3-环氧基-1′,3′-庚二烯基)-6-羟基-5-(3-甲基-2-丁烯基)苯甲醛(4)、2-(2′,3-环氧基-1′,3′5′-庚三烯基)-6-羟基-5-(3-甲基-2-丁烯基)苯甲醛(5)、2-(3E,5E-庚二烯基)-3,6-二羟基-5-(3-甲基-2-丁烯基)苯甲醛(6)、酪醇(7)、对羟基苯甲酸(8)、苔黑酚(9)。活性测试结果显示化合物3～6对3种肿瘤细胞株有较好的细胞毒活性;除化合物1和6外,其他化合物对普通变形杆菌具有明显的抑制活性,其中化合物9还对金黄色葡萄球菌和大肠杆菌表现出很好的抑菌效果。结论苯甲醛类化合物是冠突散囊菌的主要代谢产物,化合物7和9为首次从该属真菌中分离得到,化合物2、3、5、6和8均为首次从该真菌中分离得到。

Objective To study the secondary metabolites of Eurotium cristatum from Fu Brick Tea and their biological activities.Methods The compounds were isolated by various column chromatographies on normal phase silica gel,reversed silica gel,Sephadex LH-20,preparative TLC,and recrystallization.The structures were identified by the extensive analysis on their spectroscopic data.The inhibitory effects of compounds 1—9 against SF-268,MCF-7,and NCI-H460 cell lines were tested in vitro by SRB method.The inhibitory effects of compounds 2—9 on Staphyloccocus aureus,Escherichia coli,Salmonella enterica subsp.enterica,Shigella dysenteriae,and Proteus vulgaris were tested by MTT method.Results Nine compounds were isolated from the extract of liquid fermentation broth of E.cristatum in Fu Brick Tea and identified as physcion（1）,1,5-dihydroxy-3-methoxy-7-methyl-anthracene-9,10-dione（2）,flavoglaucin（3）,2-（2′,3-epoxy-1′,3′-heptadienyl）-6-hydroxy-5-（3-methyl-2-butenyl） benzaldehyde（4）,2-（2′,3-epoxy1′,3′,5′-heptatrienyl）-6-hydroxy-5-（3-methyl-2-butenyl） benzaldehyde（5）,isodihydroauroglaucin（6）,tyrosol（7）,p-hydroxybenzoic acid（8）,and orcinol（9）.The results of the bioactivity test showed that compounds 3—6 displayed the good cytotoxic acitivities against the three tumor cell lines,and all the compounds exhibited the strong inhibitory activities against P.vulgaris except compounds 1 and 6.Besides,compound 9 also showed the significant inhibitory effects against S.aureus and E.coli.Conclusion The benzaldehyde compounds are considered as the main metabolites of E.cristatum,compounds 7 and 9 are reported from the species of genus Eurotium Link： Fr.for the first time and compounds 2,3,5,6,and 8 are firstly isolated from the fungus.