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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
Subtype硕士
Thesis Advisor王斌贵
2017-05
Degree Grantor中国科学院大学
Place of Conferral北京
Degree Discipline海洋生物学
Keyword海洋真菌 次级代谢产物 抗菌活性
Abstract
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。
Other Abstract
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.
Language中文
Document Type学位论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/136642
Collection实验海洋生物学重点实验室
Affiliation中国科学院海洋研究所
Recommended Citation
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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