达托霉素生产菌(ATCC31568)的紫外诱变和抗生素抗性筛选

Ultraviolet mutagenesis and resistance selection of daptomycin-producing strain(ATCC31568)

目的建立一种提高达托霉素生产水平的方法。方法以玫瑰孢链霉菌(ATCC31568)为出发菌株,利用紫外诱变结合抗生素抗性筛选以提升达托霉素生产水平。结果通过对致死率和正突变率的考察,确定紫外诱变时间为45s,利福平和庆大霉素的合适筛选浓度分别为1.4μg/mL、1.0μg/mL,得到的突变株经摇瓶发酵后,采用高效液相色谱仪(HPLC)对最终发酵液进行检测。筛选到的突变株Ge-27最高产量达到79.44 mg/L,比原始菌株提高了42.2%。结论采用紫外诱变结合抗生素抗性筛选得到了达托霉素合成水平提高的菌株。

Objective To set up an efficient method for increasing the production of daptomycin. Methods Ultraviolet mutagenesis and resistance selection was used to increase the production of daptomycin from original strain of Streptomyces roseosporus ATCC31568. Results The results showed that the dose of UV irradiation was determined 45s in term of lethality rate and positive mutation, the minimum inhibitory concentration（MIC） of the Rifampicin and Gentamicin were 1.4 μg/mL＆1.0 μg/mL, the final broth in flasks was determined by HPLC. High daptomycin-producing mutant Ge-27 was isolated in the screening experiments, and its yield of daptomycin reached 79.44 mg/L,which was 42.2% higher than its parental strain. Conclusion The mutants was obtained whose production of daptomycin could be increased by UV mutagenesis and resistance selection.