Method for preparing epothilone by inducing sorangium cellulosum expression
A technology of S. cellulosus and fiber induction, which is applied in the field of microbial fermentation of anti-tumor drugs, can solve the problems of low expression efficiency of epothilone, achieve the effects of shortening fermentation culture time, improving fermentation efficiency, and simple analysis and elution methods
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[0055] Example 1
[0056] S1. Preparation of fermentation strains: Add the lyophilized strains of Solanum cellulosum ATCC25532 into a test tube containing 3 mL of normal saline, shake evenly, and add 1 mL of the solution to a petri dish containing a solid medium for coating Evenly. Solid medium formula (each component is in weight percentage): 2% pulp liquid, 2% potato starch, 1% peptone, 2% corn steep liquor, 0.2% dipotassium hydrogen phosphate, 2% agar powder, pH 7.0, After sterilizing at 121°C for 20 minutes, add 20 mg kanamycin B per liter of medium to the ultra-clean workbench, mix well, and pour it into a petri dish. After incubating for 48 hours in a 28°C incubator. Select the uniformly growing, orange colony moss, scrape it and plant it in a 500mL Erlenmeyer flask containing 100mL liquid medium, and place it in a constant temperature gas bath shaker with a temperature of 28℃ and a rotating speed of 220rpm for 72 hours, when the biomass reaches 10 9 CFU / mL is used as a ...
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[0062] Example 2
[0063] S1. Preparation of fermentation strains: Add the lyophilized strains of Solanum cellulosum ATCC25532 into a test tube containing 3 mL of normal saline, shake evenly, and add 1 mL of the solution to a petri dish containing a solid medium for coating Evenly. Solid medium formula (each component is in weight percentage): 1% pulp liquid, 1% potato starch, 1% peptone, 1% corn steep liquor, 0.1% dipotassium hydrogen phosphate, 1.5% agar powder, pH 7.0, After sterilizing at 121°C for 20 minutes, add 15mg kanamycin B per liter of medium on the ultra-clean workbench, mix well, and pour it into a petri dish. After incubating for 40 hours in a 25°C incubator. Select the uniformly growing, orange colony moss, scrape it and plant it in a 500mL Erlenmeyer flask containing 100mL liquid medium, and place it in a constant temperature air-bath shaker with a temperature of 25℃ and a rotating speed of 240rpm for 60 hours. When the biomass reaches 10 9 CFU / mL is used as a...
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[0069] Example 3
[0070] S1. Preparation of fermentation strains: Add the lyophilized strains of Solanum cellulosum ATCC25532 into a test tube containing 3 mL of normal saline, shake evenly, and add 1 mL of the solution to a petri dish containing a solid medium for coating Evenly. Solid medium formula (each component is in weight percentage): 5% pulp liquid, 2% potato starch 5% peptone, 2% corn steep liquor, 0.2% dipotassium hydrogen phosphate, 2% agar powder, pH 7.0, After sterilizing at 121°C for 20 minutes, add 30 mg kanamycin B per liter of medium on the ultra-clean workbench, mix well, and pour it into a petri dish. After incubating for 50 hours in a 30°C incubator. Select the uniformly growing, orange colony moss, scrape it and plant it in a 500mL Erlenmeyer flask containing 100mL liquid medium, and place it in a constant temperature air-bath shaker with a temperature of 28℃ and a rotation speed of 260rpm for 80 hours, when the biomass reaches 10 9 CFU / mL is used as a f...
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