两株植物内生真菌Talaromyces purpureogenus和Penicillium rapier中次级代谢产物的

发布时间：2018-01-17 12:28

本文关键词：两株植物内生真菌Talaromyces purpureogenus和Penicillium rapier中次级代谢产物的研究　出处：《河北大学》2017年硕士论文　论文类型：学位论文

【摘要】：近年来,由于人工合成的药物可能导致组织残留、抗药性增强等负面影响,天然药物的开发成为现今人们关注的焦点。内生真菌与宿主植物建立的和谐共生的关系,使其次级代谢产物具有结构复杂多样,生物活性广泛等特点,这为发掘先导化合物和新药研发奠定了物质基础,在工农业及生物制药等领域也存在广阔的应用前景。本论文以药用植物内生真菌为研究对象,开展次级代谢产物的结构以及生物活性的研究。研究内容主要包括:内生真菌的分离与筛选、内生真菌次级代谢产物的分离纯化以及结构鉴定、部分化合物的生物活性筛选。对采自河北省石家庄地区的10株药用植物组织进行内生真菌分离鉴定,共分离得到37株真菌,并通过小规模发酵筛选出两株真菌,三七内生真菌Talaromyces purpureogenus和艾草内生真菌Penicillium rapier作为本论文的研究对象,对其菌株发酵产物利用萃取、薄层色谱、硅胶柱色谱、ODS柱色谱、Sephadex LH-20凝胶柱色谱以及半制备高效液相等分离手段,NMR、IR、UV、MS、CD等波谱学技术,分别从三七内生真菌Talaromyces purpureogenus和艾草内生真菌Penicillium rapier中分离鉴定了18个(1-18)和12个(19-30)单体化合物,其中2个新化合物(1-2)。在构型确定方面,我们采用ECD、OR并结合化学实验首次鉴定了Drimane倍半萜类化合物(1-2)的绝对构型。将分离得到的部分化合物采用微量稀释的方法对14株革兰氏阳性和阴性致病细菌进行了抑菌活性筛选,发现化合物对细菌有一定的抑制活性,化合物4对枯草芽孢杆菌具有很好的抑制活性,MIC值为1.56μM;化合物5对溶壁微球菌和副溶血性弧菌具有比阳性对照环丙沙星低几倍的抑制活性,MIC值分别为0.73、0.18μM,具有开发抗菌类药物的潜力。对分离得到的部分化合物用DPPH法进行抗氧化活性测试,结果表明化合物无明显抗氧化活性。综上所述,本研究从三七内生真菌Talaromyces purpureogenus和艾草内生真菌Penicillium rapier中分离获得了具有较好活性的天然产物,运用ECD、OR以及量子化学计算的方法确定了化合物的绝对构型。活性筛选表明,部分化合物具有较好的抗菌活性,这为药物先导化合物的开发和应用研究提供了化合物基础。
[Abstract]:In recent years, synthetic drugs may lead to tissue residues, increased resistance and other negative effects. The development of natural drugs has become the focus of attention. The harmonious symbiotic relationship between endophytic fungi and host plants makes their secondary metabolites have the characteristics of complex structure and wide biological activity. This has laid a solid foundation for the discovery of leading compounds and new drugs, and has a broad application prospect in the fields of industry, agriculture and biopharmaceutics.In this paper, endophytic fungi of medicinal plants are taken as the research object. Studies on the structure and biological activity of secondary metabolites were carried out. The main contents of the study included: isolation and screening of endophytic fungi, separation and purification of secondary metabolites of endophytic fungi and identification of their structures. Isolation and identification of endophytic fungi from 10 medicinal plant tissues collected from Shijiazhuang area of Hebei Province. A total of 37 strains of fungi were isolated. Two strains of fungi were screened by small-scale fermentation. The endophytic fungi Talaromyces purpureogenus and Penicillium rapier of Panax notoginseng were studied in this paper. The fermentation products of the strain were separated by extraction, thin layer chromatography, silica gel column chromatography and Sephadex LH-20 gel column chromatography. Spectroscopic techniques, such as IRU / UV / MS / CD. 18 strains were isolated from the endophytic fungi Talaromyces purpureogenus and Penicillium rapier of Panax notoginseng. 1-18) and 12 monomers. Two of the new compounds were identified by ECD. Drimane sesquiterpenes were identified by OR and chemical experiments for the first time. The antimicrobial activity of 14 Gram-positive and negative pathogenic bacteria was screened by microdilution method. It was found that the compound had a certain inhibitory activity on bacteria, and compound 4 had a good inhibitory activity against Bacillus subtilis. The MIC value of compound 4 was 1.56 渭 m. The inhibitory activity of compound 5 on micrococcus and Vibrio parahaemolyticus was several times lower than that of ciprofloxacin positive control, and the MIC value of compound 5 was 0.73 渭 M and 0.18 渭 M. respectively. It has the potential to develop antimicrobial drugs. The antioxidant activity of some of the isolated compounds was tested by DPPH method. The results showed that the compounds had no obvious antioxidant activity. In this study, endophytic fungi Talaromyces purpureogenus and Penicillium from Panax notoginseng were studied. The natural products with good activity were isolated from rapier. The absolute configuration of the compounds was determined by means of ECDOR and quantum chemical calculation. The screening of activity showed that some of the compounds had better antibacterial activity. This provides the basis for the development and application of drug leading compounds.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R915

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