【作者】 肖永堂；

【导师】 宋思扬；

【作者基本信息】 厦门大学 ， 微生物学， 2006， 硕士

【摘要】 本文对采集自浙江舟山海域底泥的72株海洋真菌、福建省九龙江口及云霄县红树林的116株红树林共附生真菌,进行了杀小杆线虫、杀卤虫和乙酰胆碱酯酶(AChE)抑制活性的初步研究。对筛选出具有较强AChE抑制活性的#229真菌和抗菌活性的C-3-5-9真菌的代谢产物进行了初步的研究。我们对188株真菌粗提物的杀小杆线虫活性进行测试,结果表明,对小杆线虫有一定的毒杀作用。在0.5mg/mL的浓度下,毒力级别达到A级的菌株数占1.60%;而致50%以上死亡率菌株数占43.1%。经复筛,#164真菌粗提物具有较强的杀小杆线虫活性,0.5mg/mL浓度下处理24h,小杆线虫死亡率为96.67%。对188株真菌粗提物的杀卤虫活性进行研究,其中#122真菌粗提物的活性较强,在0.5mg/mL浓度下,处理2h后卤虫死亡率就达到了100%。另外,#13、#45、#83、#123、#305等6株真菌粗提物对卤虫具有较强的毒杀作用。在0.5mg/mL浓度下,被测试卤虫24h死亡率均在85%以上。进一步考察上述6株真菌粗提物浓度梯度与杀卤虫活性关系。结果表明,#305真菌粗提物杀卤虫活性最强,在100μg/mL的浓度下,处理24h,卤虫死亡率高达90%,其LC50仅为75μg/mL。从188个粗提物中筛选到#229真菌和#170真菌粗提物具有较强的AChE抑制活性,IC50分别是30μg/mL和25μg/mL。进一步实验结果表明,#229真菌和#170真菌很可能是同一个菌种,因为它们代谢产物的TLC结果完全相同。对具有AChE抑制活性的#229真菌的代谢产物进行活性跟踪分离,从中分离出具有高效AChE抑制活性的化合物territrem B,即土震素B。另外,从该真菌的代谢产物分离到了具有抗真菌活性的聚酮类化合物palitantin,该化合物对白色假丝酵母(Candida albicans)的MIC值为62.5μg/mL。对初筛具有抗菌和杀卤虫活性的C-3-5-9真菌的代谢产物进行分离。从中分离到了具有较强肿瘤细胞毒活性的化合物gliotoxin和acetylgliotoxin。Acetylgliotoxin对MG-63细胞株的IC50小于0.75μg/mL,对B淋巴瘤Raji细胞的IC50介于1μg/mL和2μg/mL之间,2μg/mL时,对肝癌细胞HEPG的抑制率达95%,表现出较强体外细胞毒活性。此外,对白色假丝酵母的MIC小于19μg/mL,对黑曲霉的MIC小于39μg/mL。该化合物对枯草杆菌、大肠杆菌和更多还原

【Abstract】 This thesis covers preliminary study on the anti-Rhabditis sp., anti-brine shrimps and AChE inhibitory activity of 188 marine-derived fungi. Furthermore, we studied the metabolites of two marine-derived fungi, #229 and C-3-5-9.We tested the anti-Rhabditis sp. for crude extracts of 188 fungi. The result is metabolites of marine-derived fungi are toxic against Rhabditis sp.. At the concentration of 0.5mg/mL, the virulence of grade A accounts for 1.60% of the total strains. The strains which lead to up 50% mortality account for 43.1%. By second screening, the crude extract of #164 is most active against Rhabditis sp.. At the concentration of 0.5mg/mL, the mortality is up to 96.67% after treating for 24h. We tested the anti-brine shrimps for crude extracts of 188 fungi. The crude extract of #122 is most active. At the concentration of 0.5mg/mL, the mortality is 100% after treating for 2h. Furthermore, the crude extracts of #13、#45、#83、#123、#305 are toxic against brine shrimps. At the concentration of 0.5mg/mL, the mortalities are all above 85% after treating for 24h.We studied the relationship between concentrations of crude extracts and anti-brine shrimps for the above 6 fungi. The crude extract of #305 is most active against brine shrimps. At the concentration of 0.1mg/mL, the mortality is 90% after treating for 24h. Its LC50 is only 75μg/mL.Among the 188 crude extracts, the crude extracts of #229 and #170 are most active against AChE. Their IC50s are respectively 30μg/mL and 25μg/mL. But further study showed the two fungi are possibly the same strain. The TLC result of the crude extracts from the two fungi is the same.We studied the metabolites of #229 fungus under the guidance of bioactive direction. We isolated an active AChE inhibitor, territrem B. Furthermore, we also isolated an antifungal compound, palitantin with its MIC value against Candida albicans 62.5μg/mL.The crude extract of C-3-5-9 fungus exhibited antifungal, antibacterial and anti-brine shrimp activities. We isolated two cytotoxitic compounds, gliotoxin and更多还原