Lipstatin fermentation method and fermentation medium
A fermentation method and culture medium technology, applied in the directions of fermentation, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of unsuitability for large-scale industrial production, complex and difficult fermentation process, and high cost of fermentation raw materials, and achieve control It is convenient and simple, realizes the effect of large-scale production and improved fermentation level
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Embodiment 1
[0066] a. Plate separation and slant culture: configure the spore medium according to the following ratio: glucose 4g / L, yeast extract 4g / L, malt extract 10g / L, calcium carbonate 2g / L, agar powder 18g / L, add alkali to adjust pH to 6.75, sterilized at 115°C for 30 minutes, and made into plate medium and slant medium for later use. Dilute the spore glycerol tubes of prastastatin-producing strains stored in liquid nitrogen cryogenically with sterile saline, spread them on the above-mentioned plate culture medium, and cultivate them for 12 days at a temperature of 28°C and a humidity of 40-50%. Inoculate the slant culture medium with a single colony that is good, full of spores, and uniform in color, and continue to cultivate for 8 days; select the slant surface that is completely covered with a uniform, white and dense bacterial layer from the above mature test tube slant and place it at 4 °C. Store in the refrigerator.
[0067] b. Slant shake flask fermentation screening of str...
Embodiment 2
[0075] Plate separation and slant culture, bacterial strain slant shake flask fermentation screening, shake flask seed culture, primary seed tank culture and secondary seed tank culture medium and culture conditions are the same as in Example 1, fermenter medium formula and dissolved oxygen rebound Pre-cultivation conditions are the same as in Example 1, the dissolved oxygen DO rebound time is 29h, after the rebound, adjust the air volume and rotating speed to control the dissolved oxygen according to the method in Example 1, and the dissolved oxygen starts to drop to zero at 89h after fermentation, and water is replenished once at 89h, 113h and 137h. The volume of water added each time is 5% of the volume of the fermentation broth, the fermentation period is 161 hours, and the tank titer is 13570ug / ml.
Embodiment 3
[0077] Plate separation and slant culture, bacterial strain slant shake flask fermentation screening, shake flask seed culture, medium and culture conditions of one-level seed tank culture are the same as embodiment 1, and the culture medium in the second-level seed tank and fermenter culture process will be Sunflower oil is replaced by an equivalent amount of soybean oil, the secondary seed tank culture conditions are the same as in Example 1, the culture conditions in the fermenter before the dissolved oxygen rebound are the same as in Example 1, the dissolved oxygen DO rebound time is 32h, and after the rebound, follow the method in Example 1 Adjust the air volume and speed to control the dissolved oxygen. Ferment until 116h and the dissolved oxygen starts to drop to zero. Water is replenished once at 116h, 140h and 164h. The volume of each replenishment is 5% of the volume of the fermentation broth. The potency of 187h in the fermentation cycle is 11890ug / ml.
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