【作者】 鞠培殿；

【导师】 刘新利；

【作者基本信息】 山东轻工业学院 ， 发酵工程， 2010， 硕士

【摘要】 粘细菌是一种革兰氏阴性、单细胞、杆状细菌,有独特的形态分化过程和社会性行为,因其能够产生广谱次级代谢产物而被人们所熟知,已经成为继放线菌、芽孢杆菌之后的第三大药源菌。本文对粘细菌菌株Corallococcus coralloides 085B04的生物学特性及其次级代谢产物进行了研究。通过显微观察和发酵培养等方法,掌握了粘细菌的生长习性及生活史,获得了粘细菌发酵液的乙酸乙酯萃取物。使用高效液相色谱法对乙酸乙酯萃取的粗产物和培养基的乙酸乙酯萃取物进行对比,通过分析发现,粗产物的色谱峰比VY-2培养基多12个。将发酵液粗提物通过硅胶色谱(石油醚/丙酮)梯度洗脱分离后,多次使用硅胶色谱、凝胶色谱和HPLC等技术对各组分进行分离纯化,最终从中分离得到粘细菌次级代谢产物19个。通过1H-NMR,13C-NMR、二维核磁数据、质谱、红外和紫外数据分析,对分离到的化合物进行了结构确证,其中化合物2、15和19是新化合物,它们分别是1-(1H-pyrrolo[3,2-c]quinolin-4-yl)ethanone ,5-(hydroxyimino)-1,3- dimethyl-1H-pyrrol-2(5H)-one和8-(色原烷-4’-甲酰)-环(S-脯氨酸-R-亮氨酸)。通过MTT法对得到的部分化合物作了抗肿瘤活性分析,得到了1个对前列腺癌细胞(PC3)和Hela YFP细胞抗性较好的化合物16,其IC50分别为7.8μg/mL和10.3μg/mL;1个对乳腺癌细胞(MCF-7)细胞抗性较好的化合物18,其IC50为13.7μg/mL。其中某些化合物,可能对其它肿瘤细胞具有很好的抗性,但需要进一步的研究。更多还原

【Abstract】 The myxobacteria are Gram-negative, unicellular bacteria with rod-shaped vegetative cells, which have the unique morphological differentiation and amazing social behaviors. Besides the readily established microbial secondary metabolite producers actinomycetes and bacillus, myxobacteria have become the third largest one, which are known for their ability to produce a broad spectrum of secondary metabolites and form fruiting bodies upon starvation.In this thesis, the biological characteristics and secondary metabolites of Corallococcus coralloides 085B04 have been studied. By the methods of microscopic observation and fermentation, the growth characters and life cycle of myxococcus have been discovered, and the acetic ether extracts of the myxobacterial fermentation broth has been obtained.It is found that there are 12 more peaks in residue than that in acetic ether extracts of the VY-2 medium, by camparing with their HPLC procedure.Separation of the crude residue by column chromatography over Silica gel by consecutively employing petroleum ether and acetone as eluents gave some fractions. Then, fractions were isolated and purificated by Silica gel chromatography, Sephadex LH-20 gel chromatography and HPLC techniques. Finaly, 19 metabolites of the myxobacteria were obtained. The chemical structures were confirmed by 1H-NMR, 13C-NMR, 2D-NMR, MS, IR and UV. Compounds 2, 15 and 19 are novel, which are named 1-(1H-pyrrolo[3,2-c]quinolin-4-yl)ethanone, 5-(hydroxyimino)-1,3-dimethyl -1H–pyrrol-2(5H)-one and 8-(chroman-4’-yl)-cyclo(S) -Pro-(R)-Leu, respectively.The antitumor bioactivities of the prepared samples were evaluated with the MCF-7, Pc3 and YFP cell lines using a MTT assay. The results show that, compound 18 exhibited moderate activity with IC50 13.7μg/mL against MCF-7, and compound 16 exhibited high cytotoxicity against human tumor PC3 and YFP cell lines, with their IC50 7.8μg/mL and 10.3μg/mL, respectively. Moreover, some of these compounds may exhibite more significantly against other tumor cells, and this project needs more further study.更多还原