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

【摘要】： 本文采用瓊脂擴(kuò)散法從29株土壤放線菌中篩選出了2株具有較強(qiáng)抗真菌活性的菌株(P-127及1043)。采用多相分類方法對(duì)放線菌P-127及1043進(jìn)行了鑒定,結(jié)果表明：菌株P(guān)-127的16SrDNA序列(獲得的gene bank登記號(hào)為EU797798)與Streptomyces padanus有著最相近的進(jìn)化關(guān)系,序列完全相同(同源性達(dá)100%),因此確定菌株P(guān)-127為Streptomyces padanus;菌株1043的16S rDNA(獲得的gene bank登記號(hào)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(duì)P-127及1043的基本發(fā)酵條件進(jìn)行了考察。通過杯碟法檢測(cè)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(guān)-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(duì)菌株P(guān)-127及1043發(fā)酵液中的活性物質(zhì)進(jìn)行了熱、酸堿穩(wěn)定性研究；針對(duì)P-127菌體的活性代謝產(chǎn)物進(jìn)行了pH紙層析、八種溶劑系統(tǒng)紙色譜試驗(yàn),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(duì)P-127菌體的活性代謝產(chǎn)物進(jìn)行分離,得到粗品。再利用制備型高效液相進(jìn)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(cè)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(xiàn)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(xué)位授予單位】：沈陽藥科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R379

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本文編號(hào)：1912548