本文選題：共生菌 + 次生代謝產(chǎn)物��； 參考：《揚(yáng)州大學(xué)》2014年碩士論文

【摘要】：共生菌在與宿主長期協(xié)同進(jìn)化的過程中,往往擁有比較特殊的代謝途徑,能夠產(chǎn)生豐富的結(jié)構(gòu)多樣性和生物學(xué)活性多樣性的次生代謝產(chǎn)物。共生菌還具有可再生性,且生長周期短、代謝易于控制、菌種易于選育等諸多優(yōu)勢,因此,共生菌來源的天然產(chǎn)物研究越來越受到人們的重視。 菌株IFB-WQ是南京大學(xué)功能生物分子研究所從白茅(Imperata cylindrical)中分離獲得的一株內(nèi)生真菌,根據(jù)形態(tài)學(xué)特征,鑒定為球毛殼菌(Chaetomiun globosum)。本論文課題對該菌株固體發(fā)酵產(chǎn)物的化學(xué)成分進(jìn)行了研究。球毛殼菌IFB-WQ固體發(fā)酵產(chǎn)物經(jīng)氯仿／甲醇(v/v1：1)混合溶劑浸提數(shù)次,減壓去除溶劑,除油后得粗浸膏26g。粗浸膏經(jīng)硅膠柱粗分得各極性組分；以1H-NMR譜和TLC示蹤,組合使用硅膠柱層析、凝膠柱層析、高效液相色譜等方法對各組分進(jìn)行分離純化獲得6個(gè)化合物。采用高分辨質(zhì)譜、1D和2D核磁共振譜等現(xiàn)代波譜技術(shù)及文獻(xiàn)比對的方法對化合物進(jìn)行結(jié)構(gòu)鑒定,上述化合物分別被鑒定為Chaetoglobosin F (1)、Chaetoglobosin Fex (2)、Chaetoglobosin E (3)、Cytoglobosins A (4)、Penochalasin C (5)和isochaetoglobosin D (6),它們皆為細(xì)胞松弛素類化合物。 菌株IFB-WQ2是南京大學(xué)功能生物分子研究所從海螺(Busycon canaliculatu)中分離獲得的一株放線菌,根據(jù)形態(tài)學(xué)特征,鑒定為鏈霉菌(Streptomyces sp.)。本研究對該菌株固體發(fā)酵產(chǎn)物的化學(xué)成分進(jìn)行了分離和鑒定。放線菌IFB-WQ2固體發(fā)酵產(chǎn)物經(jīng)氯仿／甲醇(v/v1：1)混合溶劑浸提數(shù)次,減壓去除溶劑,除油后得粗浸膏35g。粗浸膏經(jīng)硅膠柱粗分得各極性組分,以1H-NMR譜和TLC示蹤,組合使用硅膠柱層析、凝膠柱層析、高效液相色譜等方法對各組分進(jìn)行分離純化獲得4個(gè)化合物。采用高分辨質(zhì)譜、1D和2D核磁共振譜等現(xiàn)代波譜技術(shù)及文獻(xiàn)比對的方法對化合物進(jìn)行結(jié)構(gòu)鑒定,它們分別被鑒定為己六醇(7)、胡蘿卜苷(8)、2-oxo-3-Pyrrolidinecarboxylic acid (9)和麥角甾醇(10)。 采用MTT法測定了化合物1、3和5對人肝癌細(xì)胞株SMMC-7721和人宮頸癌細(xì)胞株HeLa的體外細(xì)胞毒活性,結(jié)果發(fā)現(xiàn)：化合物1、3和5對SMMC-7721細(xì)胞均具有一定的細(xì)胞毒活性,其IC50值分別為49.2μg/ml (0.092μmol/ml)、51.7μg/ml (0.097μmol/ml)和35.4μg/ml (0.069μmol/ml);化合物1、3和5對HeLa細(xì)胞也均具有一定的細(xì)胞毒活性,其IC50值分別為52.7μg/ml (0.099μmol/ml)、12.6μg/ml (0.024μmol/ml)和43.9μg/ml (0.086μmol/ml)。此外,還開展了化合物1-6的體外清除DPPH抗氧化應(yīng)激活性研究,結(jié)果顯示：化合物1和6具有很強(qiáng)的抗氧化活性,化合物1的IC50值為0.002μmol/ml,化合物6的IC5o值為0.002μmol/ml,化合物2-4具有弱的抗氧化活性,化合物5無抗氧化活性。天然產(chǎn)物是藥物先導(dǎo)化合物的豐富來源,本研究從白茅內(nèi)生菌C. globosum IFB-WQ中分離獲得的細(xì)胞松弛素類化合物既具有體外抗腫瘤活性,又具有顯著的體外抗氧化活性,值得深入研究。
[Abstract]:In the process of long-term co-evolution with the host, symbiotic bacteria often have a special metabolic pathway, which can produce abundant secondary metabolites of structural diversity and biological activity diversity.Symbiotic bacteria also have many advantages such as reproducibility, short growth cycle, easy control of metabolism, easy selection and so on. Therefore, people pay more and more attention to the study of natural products from symbiotic bacteria.Strain IFB-WQ is an endophytic fungus isolated from Imperata Cylindricalus from the Institute of functional Biomolecular, Nanjing University. According to its morphological characteristics, it was identified as Chaetomiun globosumsuma.In this paper, the chemical constituents of the solid fermentation products of the strain were studied.The solid fermentation product of IFB-WQ was extracted with chloroform / methanol v / v 1: 1 mixed solvent for several times, then the solvent was removed by decompression, and the crude extract was obtained after removing oil.Six compounds were isolated and purified by silica gel column chromatography, gel column chromatography and high performance liquid chromatography with 1H-NMR spectrum and TLC tracer.The structures of the compounds were identified by means of modern spectral techniques such as high resolution mass spectrometry 1D and 2D nuclear magnetic resonance spectroscopy and literature comparison.The above compounds were identified as Chaetoglobosin F ~ (1) and isochaetoglobosin D ~ (6), which are cytochalasins.The strain IFB-WQ2 was isolated from Busycon canaliculatu by the Institute of functional Biomolecular, Nanjing University. It was identified as Streptomyces sp.In this study, the chemical constituents of the solid fermentation product of the strain were isolated and identified.The solid fermentation product of actinomycetes IFB-WQ2 was extracted with chloroform / methanol v / v 1: 1) mixed solvent several times, the solvent was removed by decompression, the crude extract 35 g was obtained after removing oil.Four compounds were isolated and purified by silica gel column chromatography, gel column chromatography and high performance liquid chromatography with 1H-NMR spectrum and TLC tracer.The structures of the compounds were identified by modern spectroscopic techniques such as 1D and 2D NMR spectra and literature comparison. They were identified as hexadecanol 7, daucoside 2 oxo-3-Pyrrolidinecarboxylic acid 9) and ergosterol 1010, respectively.The cytotoxic activities of compounds 1t3 and 5 on human hepatoma cell line SMMC-7721 and human cervical cancer cell line HeLa in vitro were determined by MTT assay. The results showed that compounds 1t3 and 5 had cytotoxic activity to SMMC-7721 cells.Their IC50 values were 49.2 渭 g/ml (0.092 渭 mol / ml) and 35.4 渭 g/ml (0.069 渭 mol / ml), respectively, and their IC50 values were 52.7 渭 g/ml 0.099 渭 mol / ml 12.6 渭 g/ml 0.024 渭 mol / ml and 43.9 渭 g/ml 0.086 渭 mol / ml, respectively.In addition, the anti-oxidative stress activity of compounds 1-6 was also studied in vitro. The results showed that compounds 1 and 6 had strong antioxidant activity.The IC50 value of compound 1 was 0.002 渭 mol / ml, and the IC5o value of compound 6 was 0.002 渭 mol / ml. Compound 2-4 had weak antioxidant activity, while compound 5 had no antioxidant activity.Natural products are rich sources of drug leading compounds. The cytochalasins isolated from C. globosum IFB-WQ in this study not only have antitumor activity in vitro, but also have obvious antioxidant activity in vitro, which is worthy of further study.
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：Q936

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