Process for selecting cultivating high yield strain of datomycin by laser inducing rose sporestreptomycete
A technology of Streptomyces roseospora and daptomycin, which is applied in the field of breeding daptomycin high-yielding mutants by means of laser mutagenesis, can solve unseen problems and achieve the effects of simple equipment, easy method and safe operation.
Examples
Embodiment 1
[0013] At room temperature, take Streptomyces roseospora slant and make 10 with sterile water 4 ~10 5 spores / ml of spore suspension, draw 0.2ml and place it in a sterile quartz tank with a total volume of 3.5ml (10×10×35mm), and carry out He-Ne laser (λ=632.8nm, irradiation distance 20cm, laser beam diameter 10mm) ) for 5 minutes of irradiation, with an irradiation power of 10mW. Take 0.1ml of the irradiated suspension and dilute it by 100-1000 times, spread it on Gaoshi No. 1 medium plate, cultivate it at 30°C for 10 days, pick a single strain with good growth and inoculate it in 30ml of liquid fermentation medium (composition For: glucose 0.75g / L, dextrin 3g / L, casein 1g / L, peanut powder 0.5g / L, L-asparagine 0.1g / L, K 2 SO 4 0.5g / L, pH 7.5) in 250ml shake flasks, at 30°C, 200rpm shaker culture 110h, HPLC method detection of daptomycin production in the fermentation broth, selection of genetically stable mutants with high yield of daptomycin. The positive mutation rate o...
Embodiment 2
[0015] At room temperature, take Streptomyces roseospora slant and make 10 with sterile water 4 ~10 5 spores / ml of spore suspension, draw 0.2ml and place it in a sterile quartz tank with a total volume of 3.5ml (10×10×35mm), and carry out He-Ne laser (λ=632.8nm, irradiation distance 20cm, laser beam diameter 10mm) ) for 25 minutes of irradiation, with an irradiation power of 30mW. Take 0.1ml of the irradiated suspension and dilute it by 100-1000 times, spread it on Gaoshi No. 1 medium plate, cultivate it at 30°C for 10 days, pick a single strain with good growth and inoculate it in 30ml of liquid fermentation medium (composition For: glucose 0.75g / L, dextrin 3g / L, casein 1g / L, peanut powder 0.5g / L, L-asparagine 0.1g / L, K 2 SO 4 0.5g / L, pH 7.5) in 250ml shake flasks, at 30°C, 200rpm shaker culture 110h, HPLC method detection of daptomycin production in the fermentation broth, selection of genetically stable mutants with high yield of daptomycin. The positive mutation rate ...
Embodiment 3
[0017] At room temperature, take Streptomyces roseospora slant and make 10 with sterile water 4 ~10 5 spores / ml of spore suspension, draw 0.2ml and place it in a sterile quartz tank with a total volume of 3.5ml (10×10×35mm), and carry out He-Ne laser (λ=632.8nm, irradiation distance 20cm, laser beam diameter 10mm) ) for 30 minutes of irradiation, with an irradiation power of 25mW. Take 0.1ml of the irradiated suspension and dilute it by 100-1000 times, spread it on Gaoshi No. 1 medium plate, cultivate it at 30°C for 10 days, pick a single strain with good growth and inoculate it in 30ml of liquid fermentation medium (composition For: glucose 0.75g / L, dextrin 3g / L, casein 1g / L, peanut powder 0.5g / L, L-asparagine 0.1g / L, K 2 SO 4 0.5g / L, pH 7.5) in 250ml shake flasks, at 30°C, 200rpm shaker culture 110h, HPLC method detection of daptomycin production in the fermentation broth, selection of genetically stable mutants with high yield of daptomycin. The positive mutation rate ...
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Description
Claims
Application Information
- IPC
- C12N13/00; C12N1/20; C12R1/465
- Inventors
- 闻建平; 卢文玉