炭样小单孢菌JXNU-1广谱抗生素产物的分离及其理化性质

Isolation and Purification of Antibiotic from the Fermentation Broth of Micromonospora carbonacea JXNU-1 and Its Physical-chemical Properties

摘要:

基于实验室自行分离到的一株具有广谱抗菌活性的炭样小单孢菌JXNU-1, 研究了该菌株广谱抗生素产物的分离纯化工艺及其部分理化性质。结果表明, 该菌产生的抗生素在中性和碱性条件下具有极高的热稳定性, 但在酸性条件下不稳定。发酵液通过离心、过滤的预处理过程后, 抗生素可吸附到717型阴离子交换树脂上, 经2 mol/L的NaCl溶液洗脱除杂, 再用20%的乙醇洗脱, 可得抗生素产物, 且在20%的乙醇溶液中加入2 mol/L的NaCl, 可大大提高抗生素的洗脱效率。抗生素的酒精——盐洗脱液经减压浓缩去酒精、透析除盐后可得抗生素的水溶液。该抗生素水溶液经纸层析分析仅检测到单一活性斑点, HPLC分析表明其纯度达到99%以上。所得抗生素在Molish反应, Benedict反应和二苯胺反应中均呈现阳性, 紫外吸收具有典型的核苷类物质的光吸收特征, 推测该抗生素为一核苷类抗生素。

Abstract:

Based on the strain of Micromonospora carbonacea JXNU-1 with board-spectrum antimicrobial activity, the technology for the isolation and purification of antibiotic from the fermentation broth of the Micromonospora carbonacea, and its physical-chemical properties were studied. The results showed that, the antibiotic was stable under the condition of high temperature and alkali, but not in acid solution. After the pretreatment of centrifugation and filtration to remove the cells and lipids, the antibiotic was absorbed to negative exchange resin, and the impurity was excluded when 2 mol/L NaCl was used as primary eluent. The antibiotic could be eluted with 20% alcohol as eluent, and the eluting speed of the antibiotic was greatly accelerated as 2 mol/L NaCl was added into 20% alcohol as final eluent. Aqueous solution of the antibiotic was yielded from the alcohol-salt eluant by decompression concentration to wipe off alcohol and by dialysis to exclude salt. One active component was detected in antibiotic solution by paper chromatography, and the HPLC purity was over 99%. As the antibiotic shows positive color-forming reaction to Molish reagents, Benedict’s reagents and Diohenvlamine reagents, combined with the characteristics of absorption spectra, it is deduced that the antibiotic belongs to nucleoside antibiotics.