Semi-solid fermentation method of natamycin and natamycin extracting method
A semi-solid fermentation, natamycin technology, applied in the directions of fermentation, chemical instruments and methods, microorganism-based methods, etc., can solve the problems of instability of natamycin, affecting biological activity, and reducing extraction yield, etc. The extraction process is simplified, the yield is improved, and the material utilization rate is high.
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Embodiment 1
[0047] Use the L-80 strain of Streptomyces luteus S.gilvosporeus as the production strain, and use Gao's No. 1 medium as the slant medium, inoculate and cultivate for 7 days, until the spore pile on the surface of the slant turns from white to grayish yellow, enter 4 ℃ refrigerator. Take it out after one week in the refrigerator, and prepare the spore suspension with sterile distilled water. The spores in the spore suspension are preferably about 100 million per milliliter.
[0048] The slant spores were cultured through secondary expansion to obtain seed liquid. The formula of the seed medium is peptone 1.2%, potassium chloride 0.8%, and glucose 1.8%. Glucose is sterilized separately at 115°C for 20 minutes. Before disinfection, the pH was 7.2. After disinfection, glucose was added in an aseptic manner, and the pH after mixing was 6.9. The primary seeds are packed with 25ml of seed liquid in a 250ml Erlenmeyer flask, cultivated at 29°C, 180rpm, on a large-amplitude shaker...
Embodiment 2
[0059] The Y-42 strain of S. gilvosporeus was used as the production strain. The slant culture medium is glucose 1.2%, yeast extract 0.4%, malt extract 0.3%, peptone 0.5%, agar 2.0%, glucose is sterilized separately, and the method is the same as above. Before disinfection, the pH value was 7.2. After inoculation with spores of the Y42 strain, it was cultured at a constant temperature of 28°C for 7 days, stored in a refrigerator at 4°C for 7 days, and then the spore suspension was made with sterile distilled water.
[0060] Secondary seed expansion is still used for seed expansion cultivation, and the seed medium formula is 2.0% glucose, 0.6% peptone, 0.6% corn steep liquor, 0.8% KCL, pH7.0. Glucose is sterilized separately, and the method is the same as above. The primary seeds were cultivated in a 50L seed tank, feeding 30L, at 28°C to 29°C, with a ventilation rate of 1:0.6 (V / V) and stirring at 220rpm, and cultivated for 24 hours. The secondary seeds are carried out in a ...
Embodiment 3
[0068]This example is carried out on a semi-solid fermentation bed. The used production strain is the L-333 strain of Streptomyces luteus S. gilvosporeus. The cultivation of its slant strain and the preparation of the spore suspension are the same as in Example 2.
[0069] Seed expansion cultivation adopts three-stage expansion cultivation method. The formulation of the seed medium is: 1.0% starch, 2.0% glucose, 1.5% bean cake powder (hot pressed), 0.2% corn steep liquor, 0.5% yeast powder, 0.03% potassium dihydrogen phosphate, and 0.8% light calcium carbonate. pH7.2 before disinfection. Glucose is sterilized separately and mixed with other substances after sterilization.
[0070] 500ml Erlenmeyer flask for primary seed culture. Pack 50ml of seed culture medium in the bottle, and inoculate 5ml of spore suspension of L-333 bacterial strain. Incubate at 29°C for 16 hours on a large-amplitude shaker at 200 rpm. Secondary seed culture is carried out in 2000ml Erlenmeyer flas...
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