1株老鼠簕内生真菌Trichoderma sp.RSF-1的次级代谢产物分离及生物学活性研究

Study on the secondary metabolites and their bioactivities from the Acanthus ilicifolius derived fungi Trichoderma sp.RSF-1

目的对漳江口红树林植物老鼠簕(Acanthus ilicifolius)内生真菌Trichoderma sp.RSF-1的次级代谢产物和生物活性进行研究。方法综合利用硅胶柱色谱、凝胶柱层析及高效液相色谱(HPLC)等分离手段对菌株发酵液粗提物进行次级代谢产物的分离纯化;利用现代波谱学方法(核磁共振、质谱及旋光等)对单体化合物进行结构鉴定;对所得化合物进行细胞毒活性评价和抗菌活性筛选。结果从Trichoderma sp.RSF-1大米发酵产物中分离鉴定了13个单体化合物:包括1个丁内酯化合物1,5个聚酮类化合物2~6,1个生物碱类化合物7,2个二倍半萜类化合物8和9,3个甾体类化合物10~12及1个脂肪酸类化合物13:piterrioneⅠ(1)、nafuredin(2)、penicilloxalone A(3)、5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone(4)、5-hydroxy-7-methoxy-2,3-dimethyl-4H-chromen-4-one(5)、fusarimone A(6)、(3R,6R)-4-methyl-6-(1-methylethyl)-3-phenylmethylperhydro-1,4-oxazine-2,5-dione(7)、fusaproliferin(8)、terpestacin(9)、麦角甾醇(10)、β-谷甾醇(11)、啤酒甾醇(12)及亚油酸(13)。结论从漳江口红树林植物老鼠簕内生真菌Trichoderma sp.RSF-1中分离获得了化合物1~13,其中化合物1为新化合物,7、8和9首次从该属分离得到,化合物7对多种癌细胞系均有显著的细胞毒活性,化合物8对部分癌细胞具有中等的细胞毒活性。化合物7和13对多种致病菌包括枯草芽孢杆菌、杀鲑气单胞菌和鳗弧菌具有较强抑制作用,其最低抑菌浓度(MIC)为6.25~50.00μg/mL。

Objective To study the chemical constituents and bioactivities of Acanthus ilicifolius-derived fungi Trichoderma sp.RSF-1 collected from Zhangjiangkou mangrove national nature reserve.Methods The isolation and purification of these compounds was performed by silica gel column chromatography,gel column chromatography and high performance liquid chromatography(HPLC).Their structures were characterized by analysis of NMR,MS and ORD,etc.The cytotoxicity and antibacterial activity of these pure compounds were tested.Results Thirteen compounds were isolated from the Acanthus ilicifolius-derived fungi Trichoderma sp.RSF-1,including one butyrolactone 1,five polyketides 2?6,one alkaloid 7,two sesterterpenes 8 and 9,three steroids 10?12 and one fatty acid 13.And their structures were identified as piterrioneⅠ(1),nafuredin(2),penicilloxalone A(3),5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone(4),5-hydroxy-7-methoxy-2,3-dimethyl-4H-chromen-4-one(5),fusarimone A(6),(3R,6R)-4-methyl-6-(1-methylethyl)-3-phenylmethylperhydro-1,4-oxazine-2,5-dione(7),fusaproliferin(8),terpestacin(9),ergosterol(10),β-sitosterol(11),cerevisterol(12)and linoleic acid(13).Conclusion Thirteen compounds were isolated from Acanthus ilicifolius-derived fungi Trichoderma sp.RSF-1 collected from Zhangjiangkou mangrove national nature reserve.Compound 1 is a new compound,and compounds 7,8 and 9 were firstly isolated from the Trichoderma genus.Compound 7 displayed obvious cytotoxicity against a variety of cancer cell lines while 8 exhibited moderate cytotoxicity.Meanwhile,compounds 7 and 13 exhibited strong antibacterial activities against multiple pathogenic bacteria including Bacillus subtilis MRSA AT33591,Aeromonas salmonicida AS42,Vibrio anguillarum HUB185 with the MIC values of 6.25?50.00μg/mL.