發(fā)布時(shí)間：2018-01-17 12:28

  本文關(guān)鍵詞：兩株植物內(nèi)生真菌Talaromyces purpureogenus和Penicillium rapier中次級(jí)代謝產(chǎn)物的研究　出處：《河北大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 植物內(nèi)生真菌分離 Talaromyces purpureogenus Penicillium rapier 絕對(duì)構(gòu)型 抗菌活性

【摘要】：近年來(lái),由于人工合成的藥物可能導(dǎo)致組織殘留、抗藥性增強(qiáng)等負(fù)面影響,天然藥物的開(kāi)發(fā)成為現(xiàn)今人們關(guān)注的焦點(diǎn)。內(nèi)生真菌與宿主植物建立的和諧共生的關(guān)系,使其次級(jí)代謝產(chǎn)物具有結(jié)構(gòu)復(fù)雜多樣,生物活性廣泛等特點(diǎn),這為發(fā)掘先導(dǎo)化合物和新藥研發(fā)奠定了物質(zhì)基礎(chǔ),在工農(nóng)業(yè)及生物制藥等領(lǐng)域也存在廣闊的應(yīng)用前景。本論文以藥用植物內(nèi)生真菌為研究對(duì)象,開(kāi)展次級(jí)代謝產(chǎn)物的結(jié)構(gòu)以及生物活性的研究。研究?jī)?nèi)容主要包括:內(nèi)生真菌的分離與篩選、內(nèi)生真菌次級(jí)代謝產(chǎn)物的分離純化以及結(jié)構(gòu)鑒定、部分化合物的生物活性篩選。對(duì)采自河北省石家莊地區(qū)的10株藥用植物組織進(jìn)行內(nèi)生真菌分離鑒定,共分離得到37株真菌,并通過(guò)小規(guī)模發(fā)酵篩選出兩株真菌,三七內(nèi)生真菌Talaromyces purpureogenus和艾草內(nèi)生真菌Penicillium rapier作為本論文的研究對(duì)象,對(duì)其菌株發(fā)酵產(chǎn)物利用萃取、薄層色譜、硅膠柱色譜、ODS柱色譜、Sephadex LH-20凝膠柱色譜以及半制備高效液相等分離手段,NMR、IR、UV、MS、CD等波譜學(xué)技術(shù),分別從三七內(nèi)生真菌Talaromyces purpureogenus和艾草內(nèi)生真菌Penicillium rapier中分離鑒定了18個(gè)(1-18)和12個(gè)(19-30)單體化合物,其中2個(gè)新化合物(1-2)。在構(gòu)型確定方面,我們采用ECD、OR并結(jié)合化學(xué)實(shí)驗(yàn)首次鑒定了Drimane倍半萜類(lèi)化合物(1-2)的絕對(duì)構(gòu)型。將分離得到的部分化合物采用微量稀釋的方法對(duì)14株革蘭氏陽(yáng)性和陰性致病細(xì)菌進(jìn)行了抑菌活性篩選,發(fā)現(xiàn)化合物對(duì)細(xì)菌有一定的抑制活性,化合物4對(duì)枯草芽孢桿菌具有很好的抑制活性,MIC值為1.56μM;化合物5對(duì)溶壁微球菌和副溶血性弧菌具有比陽(yáng)性對(duì)照環(huán)丙沙星低幾倍的抑制活性,MIC值分別為0.73、0.18μM,具有開(kāi)發(fā)抗菌類(lèi)藥物的潛力。對(duì)分離得到的部分化合物用DPPH法進(jìn)行抗氧化活性測(cè)試,結(jié)果表明化合物無(wú)明顯抗氧化活性。綜上所述,本研究從三七內(nèi)生真菌Talaromyces purpureogenus和艾草內(nèi)生真菌Penicillium rapier中分離獲得了具有較好活性的天然產(chǎn)物,運(yùn)用ECD、OR以及量子化學(xué)計(jì)算的方法確定了化合物的絕對(duì)構(gòu)型�；钚院Y選表明,部分化合物具有較好的抗菌活性,這為藥物先導(dǎo)化合物的開(kāi)發(fā)和應(yīng)用研究提供了化合物基礎(chǔ)。
[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.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R915

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