海洋生物内生真菌微紫青霉、皮落青霉和聚多曲霉次级代谢产 物及其抗菌活性研究

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	海洋生物内生真菌微紫青霉、皮落青霉和聚多曲霉次级代谢产物及其抗菌活性研究
	胡雪怡
学位类型	硕士
导师	王斌贵
	2017-05
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	海洋真菌次级代谢产物抗菌活性
摘要	吲哚二萜类次级代谢产物是天然产物研究中一类较为重要的化合物，其化学结构多样，且具有很好的抗菌、抗病毒、抗肿瘤活性，日益受到人们关注。本论文对 20 世纪 70 年代到目前为止所发现的吲哚二萜类次级代谢产物进行了系统的文献调研及综述。调研发现，来源于海洋的吲哚二萜占到了总数的约 31%，也说明了海洋是天然产物的重要来源，并具有巨大的潜力。本论文对海葵来源的内生真菌微紫青霉（Penicillium janthinellum）AS-79 的规模发酵产物进行了系统分离，采用硅胶柱层析、凝胶柱层析、制备薄层层析、制备高效液相色谱等方式，得到 14 个化合物（PJ1-14），其中 12 个为吲哚二萜类次级代谢产物（PJ1-12）， 3 个为新结构化合物（PJ1-3），通过 NMR 确定其平面结构，并用 ECD 计算及单晶衍射实验确定了其绝对构型。对分离得到的单体化合物进行抗菌活性测试，其中，化合物 PJ5、PJ7 和 PJ11 对水产病原细菌铜绿假单胞菌（Pseudomonas aeruginosa）具有显著抗性，化合物 PJ5、PJ7、PJ8 和 PJ11 对人类病原菌大肠杆菌（Escherichia coli）具有显著活性，化合物 PJ2、 PJ5、PJ7、PJ9 和 PJ11 对水生病原菌副溶血弧菌（Vibrio parahaemolyticus）具有显著活性，化合物 PJ5、PJ7 和 PJ11 对溶藻弧菌（Vibrio alginolyticus）具有显著活性。本论文对海洋真菌的共培养进行了研究，对来源于松节藻科的 19 株内生真菌进行共培养筛选，采用大米培养基静置小试发酵，对其发酵产物采用 HPLC、 TLC 及 NMR 等手段进行化学筛选，比较纯培育与共培养时次级代谢产物的异同，选择有明显差异的共培养组（EN-311、EN-434）进行规模发酵。对松节藻科海藻内生真菌皮落青霉(Penicillium crustosum) EN-311、聚多曲霉（ Aspergillus sydowii）EN-434 共培养与纯培养次级代谢产物进行系统分离与鉴定，从共培养产物中分离得到一个六元内酯类新天然产物 PA3，从 EN-434 纯培养中分离得到两个新结构的没药烷型倍半萜类化合物 PA1、PA2，并通过 ECD 计算确定其绝对构型。本论文通过三株海洋真菌规模发酵产物的系统分离及鉴定，获得了结构新颖的代谢产物，丰富了海洋真菌次级代谢产物的化学多样性，并初步揭示了海洋来源内生真菌在次级代谢产物研究中的重要地位。对分离得到的单体化合物进行了抗菌活性测试，发现了抗细菌活性显著的吲哚二萜类次级代谢产物。
其他摘要	Indole diterpenes are a class of secondary metabolites which play an important role in natural product research. Their chemical structures are diverse and have good antibacterial, antiviral and antitumor activities. They have a long history and attract more and more attentions currently. In this paper, Indole diterpenes found from 1970s to now were systematically reviewed. More than 130 secondary metabolites were described and classified according to their source and structure types, and details of activities of various secondary metabolites were revealed. The study found that about 31% of the total amount of indole diterpenes were marine-derived, showing that ocean is an important source of natural products and has great potential. In this paper, the crude extracts of Penicillium janthinellum AS-79 were separated by silica gel column chromatography, gel column chromatography, preparative thin layer chromatography (pTLC) and preparative high performance liquid chromatography (pHPLC). 14 secondary metabolites were obtained (PJ1-14), of which 12 were indole diterpenes (PJ1-12). 3 new secondary metabolites (PJ1-3) were determined by NMR, calculated ECD and X-Ray crystallographic analysis. Compounds PJ5, PJ7, and PJ11 showed activity against the aquatic pathogen Pseudomonas aeruginosa. Compounds PJ5, PJ7, PJ8 and PJ11 exhibited activity against the human pathogen Escherichia coli. Compounds PJ2, PJ5, PJ7, PJ9 and PJ11 exhibited activity against the aquatic pathogen Vibrio parahaemolyticus. In this paper, 19 fungal strains isolated from Rhodomela species were screened and co-cultured. The fermentation product were detected by means of HPLC, TLC and NMR to determine the scale fermentation strain. The isolation and identification of the secondary metabolites of co-culture and pure culture of Penicillium crustosum EN-311 and Aspergillus sydowii EN-434 were accomplished. The new natural product PA3 was isolated from co-culture. Pure culture of EN-434 gained two compounds PA1 and PA2, and the absolute configuration was determined by ECD calculation. In summary, the chemical diversity of secondary metabolites was further enriched by systematic review and identification of three fungal fermentation products. It revealed the importance of endophytic fungi in the study of secondary metabolites. The antimicrobial activity of the isolated compounds was tested, and some antibacterial indole diterpenes were found.
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/136642
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
推荐引用方式 GB/T 7714	胡雪怡. 海洋生物内生真菌微紫青霉、皮落青霉和聚多曲霉次级代谢产物及其抗菌活性研究[D]. 北京. 中国科学院大学,2017.

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