【作者】 张焱；

【导师】 高文远；

【作者基本信息】 天津大学 ， 应用化学， 2007， 博士

【摘要】 积雪草酸是积雪草所含的许多五环三萜类化合物的苷元。近年来经过研究发现,这些积雪草所含苷类的活性与苷元由直接的关系。本研究以积雪草苷为底物,利用微生物转化法生产积雪草酸,用硅胶柱分离提取积雪草酸,并用核磁共振对其结构进行了确认。为了得到能对积雪草苷进行生物转化使之成为积雪草酸的菌株,我们对三种备选菌株进行了筛选。实验发现经过筛选的黑曲霉的具有转化能力。为了提高黑曲霉对积雪草苷转化生成积雪草酸的产率,实验对黑曲霉的培养条件进行了研究,确定了培养条件的初始pH值为5,培养时间为6天,底物浓度为0.5mg/mL,葡萄糖浓度8g/L,蛋白胨加入量5g/L,KH2PO4的加入量为5g/L,培养温度为28℃,MgSO4·7H2O 1g/L,MnSO4·4H2O 1g/L,NaCl 5g/L。在此基础之上,我们进行了发酵罐培养的初步研究,经过优化试验得到的结果和摇瓶发酵的结果相当。证明此方法确有可行性。深绿卷柏（Selaginella doederleinii Hieron）是一种在我国南方部分地区广泛使用的民间药材。深绿卷柏具有具有祛风、散寒、消肿、止咳之功效,临床上多用于风湿病,风寒咳嗽。深绿卷柏的主要化学成分为黄酮类化合物。本文对深绿卷柏全草的化学成分研究。采用多种分离技术对深绿卷柏的化学成分进行分离,从深绿卷柏中共分离得到20种化合物。根据其理化性质、运用¹H-NMR,¹³C-NMR等波谱方法,以及参考文献报道的数据,鉴定出其中12种成分,其中4个化合物（robustaflavone4’,4’’’-o-dimethyl ether,robustaflavone7, 4’,7’’-o-trimethyl ether,Podocarpusflavone A,3β-Acetoxysitost-5-en-7-one）为首次从该植物分离得到。用水蒸气蒸馏法提取了深绿卷柏的挥发油成分,并用GC-MS对挥发油成分进行分析,共分离鉴定出40种挥发性成分的结构。将深绿卷柏中挥发油的主要成分与报道的另外两种卷柏科植物的挥发油成分进行了比较,发现其中有非常大的差别,这些差别应该是由于物种的不同引起的,可以作为对卷柏科卷柏属不同种植物之间的亲缘关系进行判别的辅助依据。更多还原

【Abstract】 Asiatic acid is a kind of aglycone of terpenoids in Centalla asiatica. The activity of glycoside has direct relationship with aglycone in Centalla asiatica through the research in recent years. In this study, asiaticoside was used as the substrate and reacted by glycosidase from microorganism to obtain asiatic acid.And then asiatic acid was isolated by silica gel column chromatography. The structure was examined by the method of NMR.In order to get a strain which could hydrolyse the asiaticoside to asiatic acid as much as possible, 3 strains were selected to study the ability of producting glycosidase. It confirmed Aspergillus niger is the best strain to hydrolyse the asiaticoside to asiatic acid.Moreover, the cultivate condition of Aspergillus niger was researched, in order to get asiatic acid as much as possible.It confirmed the optimization cultivate condition that pH was 5.0, cultivate time was 6 days, substrate concentration was 0.5%, glucose concentration was 8g/L, peptone concentration was 5g/L, KH2PO4 concentration was 5g/L, cultivation temperature was 28℃, MgSO4·7H2O concentration was 1g/L, MnSO4·4H2O concentration was 1g/L and NaCl concentration was 5g/L.Based on these conclusion, fermentation was scaled up in a fermentor.The result of optimized experiment was correspond with the result of shake flasks. It emonstrate that conclution is valuable.Selaginella doederleinii Hieron is a civilian herb which has been used in south of china usually. As a civilian herb, S.doederleinii has the function of eliminating wind, scattering frigidity, detumescence and treating cough. Flavanoid is the major chemical ingredients in the S. doederleinii.Chemical ingredients in S. doederleinii were studied. Multitude methods were used to isolate chemical ingredients of S. doederleinii, 20 chemical compounds were obtained. 12 chemical ingredients were identified on the basis of their chemical and physical properties as well as the data of NMR, combined with information from references. And and four chemical constituents(robustaflavone4’,4’’’-o-dimethyl ether,robustaflavone7,4’,7’’-o-trimethyl ether,Podocarpusflavone A,3β-Acetoxysitost -5-en-7-one) were obtained for the first timefrom this herb among the 12 chemical ingredients. The volatile oil of Herba S. doederleinii was extracted by steam distillation and analyzed by gas chromatographymass spectrometry (GC-MS). About 40 kinds of aetherolea compounds were identified. And obvious difference was found through comparison of the essential oil of S. doederleinii and other two kinds of Selaginella, Those differences have been generated on account of the disparity of species and might used to judge the genetic relationship in Selaginell.更多还原