海洋来源温特曲霉、草酸青霉及类阿达青霉生物活性天然产物研究

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	海洋来源温特曲霉、草酸青霉及类阿达青霉生物活性天然产物研究
	李鑫
学位类型	博士
导师	王斌贵
	2016-05-18
学位授予单位	中国科学院大学
学位授予地点	北京
学位专业	海洋生物学
关键词	海洋真菌 Aspergillus Wentii Penicillium Oxalicum Penicillium Adametzioides 次级代谢产物生物活性
摘要	海洋真菌作为天然产物的一个重要来源，近年来受到的关注与日俱增。其承受着高盐、高压、低营养、低温、无光照等所谓的生命极限环境，独特的海洋环境孕育了独特的海洋微生物。海洋真菌具有微生物在天然产物研究中的共性优势，又兼具耐受海洋极端环境的特性，使其容易在特殊生境下产生和积累不同于陆地微生物的独特的次级代谢产物。本论文对不同来源的四株海洋真菌的规模发酵产物进行了系统分离、结构鉴定以及生物活性评价，包括深海来源的温特曲霉（Aspergillus wentii）SD-310、海藻来源的温特曲霉（Aspergillus wentii）EN-48和草酸青霉（Penicillium oxalicum）EN-290以及红树林来源的类阿达青霉（Penicillium adametzioides）MA-155。采用常规的硅胶柱层析、Sephadex LH-20凝胶柱层析、反相硅胶柱层析、制备薄层层析（pTLC）、制备高效液相色谱（pHPLC）和重结晶等手段分离纯化得到单体化合物，综合运用现代波谱技术（IR、UV、MS、1D-NMR和2D-NMR）从上述四株海洋真菌中分离鉴定单体化合物80个，其中21个为新化合物。从深海来源的温特曲霉（A. wentii）SD-310的发酵产物中分离鉴定20个化合物（WT1−WT20），包括12个降二萜类新化合物（WT1−WT5、WT10−WT16）和四个海松烷型二萜新化合物（WT6−WT9）；从热带马尾藻来源的内生真菌温特曲霉（A. wentii）EN-48发酵产物中分离鉴定18个化合物（WTE1−WTE18），包括1个氧杂蒽酮类新化合物（WTE1）和一个生物碱新化合物（WTE2），并利用甲基化衍生法完成了对氧杂蒽酮类化合物的结构修正；从刺松藻内生真菌草酸青霉（P. oxalicum）EN-290的MH2培养基发酵产物中分离鉴定16个化合物（OC1−OC16），包括1个生物碱类新化合物（OC1）和一个混源萜类新化合物（OC2）；从其PDB培养基发酵产物中分离鉴定13个化合物（OCP1−OCP13），包括8个混源萜类化合物；从红树林来源的类阿达青霉（P. adametzioides）MA-155的发酵产物中分离鉴定13个化合物（PD1−PD13），包括1个聚酮类新化合物（PD1）。对分离得到的部分化合物进行了抑菌、抑藻、抗氧化、细胞毒等生物活性测试。其中WT6对四株植物病害真菌烟草黑胫病菌（Phytophthora parasitica）、番茄枯萎病菌（Fusarium. oxysporum f. sp. lycopersici）、小麦赤霉病菌（Fusarium graminearum）和苹果轮纹病菌（Botryosphaeria dothidea）表现出显著抑制活性，MIC分别为8 μg/mL、4 μg/mL、1 μg/mL和4 μg/mL；化合物WTE2、WTE6、WTE7、WTE8、WTE10、WTE11和WTE12表现出不同程度的自由基清除活性，其中WTE2、WTE10以及WTE11的活性强于阳性对照；OC1对赤潮微藻新月菱形藻（Nitzschia closterium）表现出显著的选择性抑制活性，其浓度在20、10和5 mg/mL时抑藻圈分别为20、16和10 mm；化合物OC1还对金黄色葡萄菌球菌（Staphyloccocus aureus）和副溶血性弧菌（Vibrio parahaemolyticus）表现出抑制活性，MIC值分别为2.0和16.0 μg/mL。本论文通过对上述海洋真菌代谢产物的系统分离鉴定和生物活性评价，进一步揭示了海洋真菌代谢产物的多样性及其丰富显著的生物活性，为后续的先导化合物研究和微生物活性代谢产物生物合成调控等研究提供了研究基础和依据。
其他摘要	Marine-derived fungi, which are important sources for new natural products, have attracted much attention recently. They could tolerate extreme marine environmental conditions, such as high salinity, high pressure, low temperature, oligotrophic and lightless condition, and develops various special metabolic mechanisms to adapt for the special marine environment in the long run. The special environmental conditions lead marine fungi to produce special secondary metabolites which differ from those of terrestrial microorganisms. In this dissertation, the fermentation extracts of four fungal strains, Aspergillus wentii SD-310, Aspergillus wentii EN-48, Penicillium oxalicum EN-290 and Penicillium adametzioides MA-155 were systematic investigated for their chemical constituents. Some of the isolated compounds were evaluated for their bioactivities. Twenty secondary metabolites (WT1−WT20) including twelve novel tetracyclic 20-nor-ent-pimarane diterpenoid derivatives (WT1−WT5, WT10−WT16) and four new pimarane diterpenoids (WT6−WT9) were characterized from the extracts of deep-sea sediment derived fungus A. wentii SD-310. Eighteen secondary metabolites (WTE1−WTE18) including a new xanthone (WTE1) and a new benzamide derivative (methyl 4-(3,4-dihydroxybenzamido)butanoate, WTE2) were characterized from the extracts of the marine alga-derived endophytic fungus A. wentii EN-48. Furthermore, the structure of WTE1 to be revised on the basis of a methylation method. Sixteen secondary metabolites (OC1−OC16) including a new phenolic enamide, methyl (Z)-3-(3,4-dihydroxyphenyl)-2-formamidoacrylate (OC1), and a new meroterpenoid, 15-hydroxydecaturin A (OC2), were characterized from the cultivation in a MH2 culture medium of the marine alga-derived endophytic fungus P. oxalicum EN-290. In addition, thirteen secondary metabolites (OCP1−OCP13) were characterized from the cultivation in a PDB culture medium of the fungus EN-290. Thirteen secondary metabolites (PD1−PD13) including a new polyketid (PD1) were characterized from the extracts of the mangrove-derived endophytic fungus P. adametzioides MA-155. Some of the isolated compounds were evaluated for antibacterial, antifungal, algal inhibition, antioxidant and cytotoxicity activities. WT6 exhibited selective activities against Phytophthora parasitica, Fusarium. oxysporum, Fusarium graminearum, Botryosphaeria dothidea with MIC values of 8.0, 4.0, 1.0 and 4.0 μg/mL, respectively. WTE2, WTE6, WTE7, WTE8, WTE10, WTE11 and WTE12 exhibited significant scavenging activity against DPPH. OC1 showed activities against Staphyloccocus aureus and Vibrio parahaemolyticus, with MIC values of 2.0 and 16.0 μg/mL, respectively. OC1 also showed activity against HAB causative species Nitzschia closterium with inhibition zones of 20, 16, and 10 mm at 20, 10, and 5 mg/mL by the agar diffusion method. This study may provide important proof to the utilization of natural products from marine fungi, and also provide the foundation of exploitation for lead compounds in drug discovery and research for biosynthesis of secondary metabolites of marine fungi.
学科领域	天然产物有机化学
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/116990
专题	实验海洋生物学重点实验室
作者单位	中国科学院海洋研究所
第一作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	李鑫. 海洋来源温特曲霉、草酸青霉及类阿达青霉生物活性天然产物研究[D]. 北京. 中国科学院大学,2016.

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