本文選題：放線菌 + 16S��； 參考：《沈陽藥科大學》2008年碩士論文

【摘要】： 本文采用瓊脂擴散法從29株土壤放線菌中篩選出了2株具有較強抗真菌活性的菌株(P-127及1043)。采用多相分類方法對放線菌P-127及1043進行了鑒定,結(jié)果表明：菌株P-127的16SrDNA序列(獲得的gene bank登記號為EU797798)與Streptomyces padanus有著最相近的進化關系,序列完全相同(同源性達100%),因此確定菌株P-127為Streptomyces padanus;菌株1043的16S rDNA(獲得的gene bank登記號為EU864307)與許多鏈霉菌的16S rDNA序列有著高度同源性,確定該菌株為一株鏈霉菌。 為了提高活性代謝產(chǎn)物的產(chǎn)量,對P-127及1043的基本發(fā)酵條件進行了考察。通過杯碟法檢測不同條件下發(fā)酵液中抗生素的產(chǎn)量,確立了最適搖瓶發(fā)酵條件：(1)菌株P-127為種齡34h,接種量10%,搖瓶裝液量40ml/250ml三角瓶,發(fā)酵周期為4天。(2)菌株1043為種齡40h,接種量10%,搖瓶裝液量50ml/250ml三角瓶,發(fā)酵周期為6天。 對菌株P-127及1043發(fā)酵液中的活性物質(zhì)進行了熱、酸堿穩(wěn)定性研究；針對P-127菌體的活性代謝產(chǎn)物進行了pH紙層析、八種溶劑系統(tǒng)紙色譜試驗,為初步確定該活性物質(zhì)的理化特性提供了依據(jù)。采用大孔吸附樹脂對P-127菌體的活性代謝產(chǎn)物進行分離,得到粗品。再利用制備型高效液相進一步分離純化,獲得了抗真菌活性物質(zhì)的結(jié)晶,純度較高,經(jīng)分析型高效液相色譜法檢測含量超過97%,質(zhì)譜顯示該活性物質(zhì)的分子量為670,紫外吸收波譜顯示該活性物質(zhì)具有多烯大環(huán)內(nèi)酯類抗生素的特征吸收峰,結(jié)合該菌株的多相分類鑒定和文獻檢索,提示該活性物質(zhì)可能是fungichromin。
[Abstract]:In this paper, two strains of soil actinomycetes with strong antifungal activity were screened by Agar diffusion method. The polyphase classification method was used to identify the actinomycetes P-127 and 1043. The results showed that the 16SrDNA sequence of the strain P-127 (gene bank registration number EU797798) had the closest evolutionary relationship with Streptomyces padanus. The strain P-127 was identified as Streptomyces padanus.The 16s rDNA of strain 1043 (gene bank registration number EU864307) was highly homologous with the 16s rDNA sequence of many Streptomyces, and the strain was identified as Streptomyces. In order to increase the yield of active metabolites, the basic fermentation conditions of P-127 and 1043 were investigated. The yield of antibiotics in fermentation broth under different conditions was determined by the method of cup and dish. The optimum fermentation condition of strain P-127 was established as the seed age of 34 h, the inoculation amount of 10% and the quantity of 40ml/250ml triangle bottle. The fermentation period of strain 1043 was 40 h, the inoculation amount was 10%, and the quantity of liquid in shake bottle was 50ml/250ml triangle bottle. The fermentation period was 6 days. The thermal, acid-base stability of the active substances in the fermentation broth of strain P-127 and 1043 were studied, and the pH paper chromatography and eight solvent system paper chromatography tests were carried out for the active metabolites of P-127 bacteria. It provides a basis for the determination of the physical and chemical properties of the active substance. The active metabolites of P-127 were separated by macroporous adsorption resin and the crude products were obtained. The antifungal active substances were obtained by further separation and purification by using the preparative high performance liquid phase, and the purity of the antifungal active substances was high. The mass spectrometry showed that the molecular weight of the active substance was 670. The UV absorption spectrum showed that the active substance had the characteristic absorption peak of polyene-macrolide antibiotics. Combined with the multiphasic taxonomy and literature retrieval of the strain, it was suggested that the active substance might be fungichromin.
【學位授予單位】：沈陽藥科大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R379

【參考文獻】

相關期刊論文 前10條

1 黃知源;生物農(nóng)藥及其在我國的發(fā)展簡況[J];北方蠶業(yè);2001年03期

2 司馬依·薩依木;農(nóng)藥的危害與微生物農(nóng)藥的開發(fā)[J];化學教育;2002年11期

3 李勇,楊慧敏,李銘剛,文孟良;微生物農(nóng)藥的研究和應用進展[J];貴州農(nóng)業(yè)科學;2003年02期

4 Aszalos A ,凌年;高效液相色譜用于抗生素分析[J];國外醫(yī)學.藥學分冊;1984年01期

5 鄧定輝;生物防治技術在植物保護中的應用[J];廣西植保;2000年02期

6 沈寅初;農(nóng)用抗生素研究開發(fā)的新進展[J];國外醫(yī)藥(抗生素分冊);1998年02期

7 張寧波;;農(nóng)用抗生素研究進展[J];湖北農(nóng)業(yè)科學;2006年06期

8 宋秀杰,郝國防;生物農(nóng)藥及生物防治[J];環(huán)境保護;2002年06期

9 高英立;;開展紙層析的一些體會和改進[J];海軍醫(yī)學;1986年04期

10 歐陽諒;微生物來源的殺蟲活性物質(zhì)開發(fā)利用[J];江西農(nóng)業(yè)大學學報;1993年S4期

，

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