耐盐菊芋及其内生真菌活性脂溶成分提取分离研究

	耐盐菊芋及其内生真菌活性脂溶成分提取分离研究
	王超之 1,2
学位类型	硕士
导师	季乃云 ; 杨米娜
	2015-05-14
学位授予单位	中国科学院研究生院
学位授予地点	北京
学位专业	生物工程
关键词	菊芋内生真菌次生代谢产物甾体萜类
摘要	菊芋( Helianthus tuberosus) 又叫鬼子姜或洋姜，为菊科向日葵属的多年生草本植物，在生态、食品、能源以及化学等领域都有应用，但目前针对菊芋及其内生真菌小分子天然产物开发利用却相对薄弱。文献报道菊芋中含有种类较多的萜类和甾体类化合物，部分化合物具有良好的生物活性。本论文以海岸带盐碱地种植的耐盐菊芋为对象，研究了其枝叶、块茎中的萜类和甾体类化合物的提取分离以及一株内生真菌的次生代谢产物。提取耐盐菊芋枝叶和块茎中的脂溶性成分，设计并优化植物甾醇的制取条件，经过溶剂提取、硅胶柱层析，凝胶Sephadex LH-20柱层析和高效液相色谱法等分离纯化步骤，得到β-谷甾醇和豆甾醇化合物。采用高效液相色谱法测定所得β-谷甾醇和豆甾醇的纯度，两种化合物纯度均达到90%以上。设计并优化菊芋中萜类组分的浓缩条件，采用溶剂提取、硅胶柱层析和凝胶Sephadex LH-20柱层析浓缩萜类组分，经紫外分光光度法测定，得到的总萜类组分浓度达到30%以上，卤虫活性测定证明，浓缩后的萜类具有一定的抗虫活性。采用活性导向法筛选菊芋内生真菌，并规模发酵活性菌株，采用硅胶层析、凝胶Sephadex LH-20柱层析、制备薄层层析和高效液相色谱技术分离纯化发酵提取物得到单体化合物，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7个化合物（HT1~HT7）。通过本课题的研究，增加耐盐菊芋的开发和利用手段，充分提高菊芋的附加值，使菊芋的开发利用更加精细化。开展菊芋块茎内生真菌的研究，从内生真菌天然产物化学角度研究菊芋，开拓菊芋利用新途径、新方法。
其他摘要	Jerusalem artichoke (Helianthus tuberosus), also called devilsginger or ginger, is a perennial herbaceous plant belonging to the Helianthus genus, Asteraceae family, and it is widely applied in ecology, food, energy, chemistry etc. Although Jerusalem artichoke contains a series of terpenes and steroids, but the application of them has been rarely reported. There is also no report about the secondary metabolites of endophytic fungi in Jerusalem artichoke as far as we know. In this paper, we studied the extraction and isolation of terpenoids and steroids from leaves and tubers of Jerusalem artichoke grown in salinized lands, and an endophytic fungus of Jerusalem artichoke was also chemically investigated. Lipophilic constituents were extracted from the leaves and tubers of Jerusalem artichoke, and the isolation of β-sitosterol and stigmasterol was designed and optimized. β-Sitosterol and stigmasterol were isolated by solvent extraction, column chromatography on silica gel and Sephadex LH-20, and preparative high performance liquid chromatography. The purity of β-sitosterol and stigmasterol was determined by high performance liquid chromatography. Both compounds reached a purity of more than 90%. The concentration conditions of terpenoids by solvent extraction, column chromatography on silica gel and Sephadex LH-20 were designed and optimized, and the contents of total terpenoids were determined by UV spectrophotometric method. Concentration of terpenoids has reached more than 30%, and bioassay results showed that the concentrating terpenoids have certain insecticidal activity. One endophytic fungus was selected by bioassay screening and then fermentated. 7 compounds were isolated from the fermentation extracts by column chromatography on silica gel and Sephadex LH-20, preparative thin layer chromatography and high performance liquid chromatography. The structures of these compounds were elucidated by spectroscopic methods, such as ¹H NMR、¹³C NMR and DEPT. The research increases the development, utilization as well as added value of salt tolerance Jerusalem artichoke, and makes the development and utilization of Jerusalem artichoke more refined. Study the endophytic fungi derived from Jerusalem artichoke tubers explore new utilization of Jerusalem artichoke from the angle of natural product chemistry.
语种	中文
文献类型	学位论文
条目标识符	http://ir.yic.ac.cn/handle/133337/7914
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	1.中国科学院烟台海岸带研究所 2.中国科学院大学
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	王超之. 耐盐菊芋及其内生真菌活性脂溶成分提取分离研究[D]. 北京. 中国科学院研究生院,2015.

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