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    窿缘桉内生真菌Chaetomium sp. Eef-10的鉴定及活性成分分析

    Identification of endophytic fungus Chaetomium sp. Eef-10 from Eucalyptus exserta and analysis of its active ingredients

      摘要
    • 摘要:
      目的  确定内生真菌Eef-10的分类地位，分离和鉴定该真菌中的次生代谢产物并评价其抗细菌和抗肿瘤细胞活性，以期得到具有抗细菌和抗肿瘤活性的天然活性化合物。
      方法  内生真菌的鉴定采用形态学和分子生物学相结合的方法；次生代谢产物的分离和纯化主要采用减压硅胶柱层析、葡聚糖凝胶LH-20和半制备高效液相色谱等方法；化合物的鉴定主要依据1H NMR和13C NMR等波谱学数据以及相关的参考文献；采用MTT显色法测定了次生代谢产物对5种不同供试细菌的抑制活性；采用CCK8法测定了次生代谢产物对2种不同癌细胞的抑制活性。
      结果  从内生真菌Eef-10中分离到的3个化合物，分别鉴定为Atraric acid(化合物Ⅰ)、2, 4−二羟基−3, 6−二甲基苯甲酸乙酯(Ethyl 2, 4-dihydroxy-3, 6-dimethylbenzoate)(化合物Ⅱ)和4−甲基−5, 6−二氢−2H−吡喃−2−酮(4-methyl-5, 6-dihydro-2 H-pyran-2-one)(化合物Ⅲ)。化合物Ⅱ对5种革兰阴性细菌表现出强抑制活性，最大半数抑制质量浓度(IC50)为35.87~55.50 μg/mL，化合物Ⅰ的IC50为67.25~130.55 μg/mL，化合物Ⅲ的IC50均大于200 μg/mL。抗肿瘤细胞活性的测定结果表明，化合物Ⅱ对人肝癌细胞株Hep-G2的IC50为1.50 μg/mL，活性强于阳性对照喜树碱(IC50为3.6 μg/mL)。
      结论  从内生真菌Chaetomium sp. Eef-10中分离得到化合物Ⅰ~Ⅲ，化合物Ⅱ对桉树青枯病菌和Hep-G2细胞具有较好的抑制作用。

       

      Abstract:
      Objective  To determine the classification status of endophytic fungus Eef-10. To isolate and identify secondary metabolites of Eef-10 and evaluate their antibacterial and antitumor activities in order to obtain natural active compounds.
      Method  The endophytic fungus was identified by combining morphology and molecular biology methods. The secondary metabolites were mainly separated and purified by vacuum silica gel column chromatography, sephadex LH-20 column chromatography and semi-preparative HPLC. The compounds were identified mainly based on 1H NMR and 13C NMR spectral data as well as related references. The antibacterial activities against five different test bacteria were determined by the MTT method and the antitumor activities against two cancer cells were determined by the CCK8 method.
      Result  Three compounds were isolated from the endophytic fungus Eef-10, namely atraric acid (Compound Ⅰ), ethyl 2, 4-dihydroxy-3, 6-dimethylbenzoate (Compound Ⅱ) and 4-methyl-5, 6-dihydro-2 H-pyran-2-one (Compound Ⅲ). Compound Ⅱ displayed strong inhibitory activities against five test gram-negative bacteria, and the IC50 values were 35.87−55.50 μg/mL. The IC50 values of compound Ⅰ were 67.25−130.55 μg/mL, while compound Ⅲ had IC50 values of more than 200 μg/mL for all five bacteria. The IC50 value of compound Ⅱ for Hep-G2 was 1.50 μg/mL, which was stronger than the positive control camptothecin of 3.6 μg/mL.
      Conclusion  Compounds Ⅰ−Ⅲ were isolated from the endophytic fungus Chaetomium sp. Eef-10 and compound Ⅱ showed great antibacterial and antitumor activities on R. solanacearum and Hep-G2 tumor cells.

       

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