【摘要】 目的:获得大量红霉素合成中间产物红霉素B,并进一步研究eryK基因的活性位点。方法:通过重叠PCR将eryK基因中包括BC环在内的关键氨基酸序列删除,克隆到同源重组质粒pWHM3上,继而通过染色体同源重组方法将EryK羟基化酶基因失活,构建了缺失C-12羟基化酶活性的红色糖多孢菌突变体。结果与结论:构建了红色糖多孢菌突变菌株AK17,对发酵产物进行TLC和MS分析,结果显示突变体主要合成红霉素B。更多还原

【Abstract】 Objective:To obtain a large amount of erythromycin B and to investigate the activity site in eryK. Methods:The key sequence of the BC loop region in eryK gene was knocked out and the eryK gene with 101 bp deleted was amplified by overlapping PCR, and cloned into vector pWHM3 to construct recombinant plasmid. The Saccharopolyspora erythraea mutant AK17 was constructed through chromosomal homologous recombination technique.Results and Conclusions:The S.erythraea mutant AK17 was constructed. The results of TCL and MS analysis showed that the major fermentation product of AK17 is erythromycin B.更多还原