Vitamin B12 fermentation supplementing culture medium and feeding method
A feed medium, B12 technology, applied in the field of biopharmaceuticals, can solve the problems of weakened glycine supply and affecting the growth and synthesis of bacteria, so as to relieve substrate inhibition, improve the phenomenon of aging and emptying of bacteria, and balance the growth of bacteria in the later stage. effects on metabolism
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Embodiment 1
[0025] 1. First-level seed cultivation: the volume of seed liquid is 3L
[0026] Seed medium: Add 400-600g of sucrose, 120-150g of corn steep liquor, 40-60g of betaine, 3.5-5g of magnesium oxide, 20-30g of diammonium hydrogen phosphate, 0.5-1g of zinc sulfate, and Calcium 30-50g, 5-6 dimethylbenzimidazole 0.7-1.4g, ammonium sulfate 10-20g, potassium dihydrogen phosphate 10-20g. Sterilization treatment is carried out after the batching is completed. Sterilization conditions: pressure 0.10-0.14MPa; temperature 120-124°C; sterilization time 30 minutes. Cool and keep the pressure with sterile air, the pressure is controlled at 0.01-0.02MPa, and then under the protection of the flame, insert the cultivated Pseudomonas denitrifica mother bottle seeds into the seed medium according to the inoculation amount of 1-3%. Carry out seed culture until the OD600 of the bacteria is 11.3, the pH value is 6.08, and the culture time is 20-40 hours, and the transplanting ratio is controlled at 5...
experiment example 2
[0054] 1. First-level seed cultivation: the volume of seed liquid is 6L
[0055] Seed medium: Add 800-1200g of sucrose, 240-400g of corn steep liquor, 80-120g of betaine, 7-10g of magnesium oxide, 40-60g of diammonium hydrogen phosphate, 1-2g of zinc sulfate, and Calcium 60-100g, 5,6-dimethylbenzimidazole 1.4-3g, ammonium sulfate 20-30g, potassium dihydrogen phosphate 20-30g. Sterilization treatment is carried out after the batching is completed. Sterilization conditions: pressure 0.10-0.14MPa; temperature 120-124°C; sterilization time 30 minutes. Cool and keep the pressure with sterile air, the pressure is controlled at 0.01-0.02MPa, and then under the protection of the flame, insert the cultivated Pseudomonas denitrifica mother bottle seeds into the seed medium according to the inoculation amount of 1-3%. Carry out seed culture until the OD600 of the bacteria is 12.4, the pH value is 6.18, and the culture time is 20-40 hours, and the transplanting ratio is controlled at 5-1...
experiment example 3
[0082] 1. First-level seed cultivation: the volume of seed liquid is 60L
[0083] Seed medium: Add 6400-9600g of sucrose, 1920-3000g of corn steep liquor, 640-1000g of betaine, 56-100g of magnesium oxide, 320-500g of diammonium hydrogen phosphate, 8-12g of zinc sulfate, and Calcium 480-720g, 5,6-dimethylbenzimidazole 11.2-15g, ammonium sulfate 160-240g, potassium dihydrogen phosphate 160-240g. Sterilization treatment is carried out after the batching is completed. Sterilization conditions: pressure 0.10-0.14MPa; temperature 120-124°C; sterilization time 30 minutes. Cool and keep the pressure with sterile air, the pressure is controlled at 0.01-0.02MPa, and then under the protection of the flame, insert the cultivated Pseudomonas denitrifica mother bottle seeds into the seed medium according to the inoculation amount of 1-3%. Carry out seed culture until the OD600 of the bacteria is 12.6, the pH value is 6.12, and the culture time is 20-40 hours, and the transplanting ratio is...
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