本文選題：炭樣小單孢菌 + 核苷類(lèi)抗生素�。� 參考：《江西師范大學(xué)》2013年碩士論文

【摘要】：新抗生素產(chǎn)生菌的分離、篩選是研發(fā)新抗生素的重要途徑之一，，課題組前期研究獲得的炭樣小單孢菌所產(chǎn)抗菌單體組分經(jīng)鑒定為一新型核苷類(lèi)抗生素。該抗生素具有廣譜抗菌活性，但抗菌作用特點(diǎn)及作用機(jī)理不清楚。本研究以該抗生素和常用抗生素活性檢測(cè)靶菌分別作為試驗(yàn)材料和試驗(yàn)對(duì)象，研究抗生素的體外抗菌譜、抗菌方式、抗菌強(qiáng)度及其作用機(jī)理，為進(jìn)一步開(kāi)發(fā)炭樣小單孢菌產(chǎn)抗生素成為我國(guó)具有自主知識(shí)產(chǎn)權(quán)的新型抗生素奠定堅(jiān)實(shí)的理論基礎(chǔ)，具有重大的理論研究和實(shí)踐應(yīng)用價(jià)值。研究結(jié)果如下： （1）采用瓊脂擴(kuò)散法和液體稀釋法分別測(cè)定了炭樣小單孢菌所產(chǎn)核苷類(lèi)抗生素抗菌譜及其最低抑菌濃度和最低殺菌強(qiáng)度，表明該抗生素對(duì)常見(jiàn)的G+和G-均具有抗菌作用，為廣譜抗生素，并且對(duì)G+和G-的最低殺菌濃度（MBC）一般是最低抑菌濃度（MIC）的8～16倍。 （2）采用顯微觀察法研究了炭樣小單孢菌所產(chǎn)核苷類(lèi)抗生素處理后的G+和G-細(xì)菌形態(tài)變化，結(jié)果顯示，在最低抑菌濃度的抗生素作用下，無(wú)論是G+、G-，細(xì)胞形態(tài)都發(fā)生明顯改變，許多細(xì)胞形體變長(zhǎng)，甚至成絲狀，還有部分細(xì)胞開(kāi)始破裂，并且，細(xì)胞的個(gè)體數(shù)目明顯減少。 （3）通過(guò)比較抗生素處理前后細(xì)胞對(duì)溶菌酶的敏感性及細(xì)胞壁的含量，表明炭樣小單孢菌所產(chǎn)核苷類(lèi)抗生素的作用不影響細(xì)菌細(xì)胞壁的合成，但使細(xì)胞壁變厚。 （4）通過(guò)比較抗生素作用后細(xì)胞懸浮液的電導(dǎo)率變化，表明抗生素對(duì)細(xì)菌細(xì)胞膜的滲透性沒(méi)有明顯的影響，說(shuō)明不影響細(xì)胞膜的結(jié)構(gòu)與功能。 （5）采用孔雀石綠—磷鉬雜多酸分光光度法測(cè)定了抗生素對(duì)細(xì)胞GTP酶活性的影響，表明炭樣小單孢菌所產(chǎn)抗生素可促進(jìn)細(xì)胞GTP酶的活性，進(jìn)而增強(qiáng)了GTP酶的催化作用。 （6）采用G-250考馬斯亮藍(lán)法檢測(cè)了炭樣小單孢菌所產(chǎn)抗生素作用下G+和G-細(xì)菌細(xì)胞內(nèi)蛋白質(zhì)含量的變化，及其對(duì)大腸埃希氏菌無(wú)細(xì)胞表達(dá)系統(tǒng)體外合成蛋白質(zhì)的影響，表明炭樣小單孢菌所產(chǎn)抗生素可抑制細(xì)胞蛋白質(zhì)的合成。 從上述結(jié)果可以推斷，炭樣小單孢菌所產(chǎn)抗生素通過(guò)抑制細(xì)胞蛋白質(zhì)的合成而抑制細(xì)胞的分裂，進(jìn)而實(shí)現(xiàn)其抑/殺菌功能。但有關(guān)該抗生素抑制蛋白質(zhì)合成的具體機(jī)制還有待進(jìn)一步研究。
[Abstract]:Isolation and screening of new antibiotic producing bacteria is one of the important ways to develop new antibiotics. The antibiotic has broad-spectrum antibacterial activity, but its characteristics and mechanism are not clear. In this study, the antimicrobial spectrum, antimicrobial method, antibacterial intensity and its mechanism of antibiotics were studied by using the antibiotic and the commonly used antibiotic activity detection target bacteria as the test materials and experimental objects, respectively. It lays a solid theoretical foundation for the further development of carbon-like micromonosporium antibiotics as a new type of antibiotics with independent intellectual property rights in China, and has great theoretical research and practical application value. The results are as follows: 1) the antimicrobial spectrum of nucleosides produced by Micromonosporium carboniformis and its minimum inhibitory concentration and bactericidal intensity were determined by Agar diffusion method and liquid dilution method respectively. The results showed that the antibiotic had antimicrobial effect on both G and G-, and it was a broad-spectrum antibiotic. And the minimum bactericidal concentration (MBC) of G and G- was 816 times of the minimum inhibitory concentration (MICM). The morphological changes of G and G- bacteria were studied by microscopic observation after treatment with nucleoside antibiotics produced by Micromonospora. In the presence of antibiotics with the lowest inhibitory concentration, the morphology of the cells, no matter the G G, changed significantly, many of the cells became longer, even became filamentous, and some of the cells began to break down, and, By comparing the sensitivity of cells to lysozyme and cell wall content before and after antibiotic treatment, the results showed that the effect of nucleoside antibiotics produced by Micromonosporium carboniformis did not affect the synthesis of bacterial cell wall. But the thickening of the cell wall showed that antibiotics had no obvious effect on the permeability of bacterial cell membrane by comparing the changes of the electrical conductivity of the cell suspensions after the treatment of antibiotics. The results showed that the structure and function of cell membrane were not affected. (5) the effect of antibiotics on the activity of GTP enzyme was determined by malachite green-molybdenum heteropoly acid spectrophotometry, which indicated that the antibiotics produced by Micromonosporium Carboniformis could promote the activity of GTP enzyme. Furthermore, the catalytic activity of GTP enzyme was enhanced. (6) G-250 Coomassie brilliant blue method was used to detect the changes of protein contents in G and G- bacteria cells under the action of antibiotics produced by Micromonosporium carboniformis. And its effect on the protein synthesis of Escherichia coli acellular expression system in vitro, indicating that antibiotics produced by Micromonosporium carboniformis can inhibit the synthesis of cellular protein. The antibiotics produced by Micromonosporium carboniformis inhibit cell division by inhibiting the synthesis of cell proteins and thus achieve its inhibitory / bactericidal function. However, the mechanism of inhibition of protein synthesis by this antibiotic remains to be further studied.
【學(xué)位授予單位】：江西師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：S482.28

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