海洋藻栖木霉次生代谢调控的研究

	海洋藻栖木霉次生代谢调控的研究
	宋银平
学位类型	硕士
导师	季乃云 ; 杨米娜
	2016-05-17
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	生物工程
关键词	藻栖真菌 Algicolous Fungus 木霉 Trichoderma 次生代谢产物 Secondary Metabolite 调控 Regulation 活性 Bioactivity
摘要	木霉(Trichoderma spp.)具有丰富的次生代谢产物,包括萜类、聚酮和生物碱等,并且新的活性产物不断被发现。海洋藻栖真菌次生代谢产物的研究始于20世纪90年代,从中获得的新化合物占海洋真菌来源新化合物的20%以上,成为海洋天然产物研究的热点领域之一。海洋藻栖木霉兼有木霉、海洋木霉和藻栖真菌三者的特性,而截止2016年,仅6株藻栖木霉的次生代谢产物被研究,因此,对海洋藻栖木霉的研究有重要的意义。通过对10株海洋藻栖木霉次生代谢调控的研究发现,藻栖木霉能够在海水和淡水环境中生长,甚至在高达70.0 g/L Na Cl环境下生长。全海水(盐度3.0%,p H=8.0)的培养环境下,海洋藻栖木霉表现出更好的生长情况。尽管卤化物(30.0g/L Na Cl和86.3 mg/L Na Br)或p H(8.0)对菌株具有负调控的作用,但是至少有一个因素可以促进生长。在Na Cl(70.0 g/L)、Na Br(86.3 mg/L)、p H=8.0及全海水培养条件下,海洋藻栖木霉的代谢产物中连杂原子碳上的氢及不饱和碳上的氢的所占的比例均增加。研究发现10株海洋藻栖木霉的细胞内外的分泌物均表现出不同程度的卤虫毒性、微藻抑制活性和植物病原真菌抑制活性;同时应用潮间带的短滨螺幼体作为生物活性筛选模型,发现海洋藻栖木霉的发酵液提取物在10μg/m L的浓度下表现出明显的滨螺毒性。选取哈茨木霉dl36菌株,大规模发酵,采用硅胶层析、凝胶Sephadex LH-20柱层析、制备薄层析和高效液相色谱技术分离纯化得到单体化合物,运用现代波谱技术(1H NMR、13C NMR、DEPT)对化合物进行结构鉴定,共得到6个化合物(TH-1—TH-6)通过10株海洋藻栖木霉次生代谢的调控及哈茨木霉dl36次生代谢产物的研究,深化了对藻栖木霉次生代谢的认识,并为藻栖木霉次生代谢产物的深入探索及应用奠定了基础。
其他摘要	Multifarious secondary metabolites have been obtained from Trichoderma species of various origin, including terpenoids, polyketides, and alkaloids. Studies on the secondary metabolites of marine algicolous fungi (MAFs) began in the 1990s, and over 20% of the secondary metabolites from marine-derived fungi were produced by MAFs, which have attracted a great attention for natural product researchers. Marine algicolous Trichoderma species have the characteristics of the Trichoderma, MAFs and marine-derived Trichoderma. Until 2016, only 6 strains of marine algicolous Trichoderma were chemically studied. Thus, it is significant to further explore the secondary metabolism of marine algicolous Trichoderma species. Study on secondary metabolic regulation of marine algicolous Trichoderma species revealed that marine algicolous Trichoderma species can grow in seawater and freshwater, even at 70.0 g/L NaCl. However, under the NaCl (30 g/L) and seawater conditions, marine algicolous Trichoderma species growed better. In NaCl (70.0 g/L), NaBr (86.3 mg/L), pH = 8.0, and seawater (salinity = 3.0% and pH = 8.0) conditions, their crude extracts contain more heteroatoms and unsaturated groups (vinyl and/or aryl). The intracellular and extracellular secretions of ten strains marine algicolous Trichoderma were also found to be toxic to Artemia salina and inhibitory against microalgae and phytopathogenic fungi. Moreover, some broth extracts were evaluated to be significantly toxic to the larvae of mollusc Littorina brevicula at 10 μg/mL. Due to the high activity, Trichoderma dl36 was selected for large-scaled fermentation. As a result, six compounds were obtained by column chromatographyon silica gel and Sephadex LH-20, preparative thin layer chromatography (pTLC),and semi-preparative high performance liquid chromatography (HPLC). The structures were elucidated by spectroscopic methods, involving 1H NMR、13C NMR,and DEPT. Overall, the secondary metabolisms of ten marine algicolous Trichoderma strains and the products from one of them have been explored. The results make a contribution to knowing the secondary metabolisms of marine algicolous Trichoderma species, and lay the foundation for further exploring it.
语种	中文
文献类型	学位论文
条目标识符	http://ir.yic.ac.cn/handle/133337/17533
专题	中国科学院烟台海岸带研究所知识产出_学位论文
作者单位	中国科学院烟台海岸带研究所
第一作者单位	中国科学院烟台海岸带研究所
推荐引用方式 GB/T 7714	宋银平. 海洋藻栖木霉次生代谢调控的研究[D]. 北京. 中国科学院大学,2016.

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