Method for preparing nuclease P1 through penicillium citrinum semi-continuous fermentation
A technology of Penicillium citrinum and nuclease, which is applied in the field of semi-continuous fermentation of Penicillium citrinum to produce nuclease P1, which can solve the problems of long time consumption and achieve the effect of reducing production cost and shortening the fermentation working hours
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Embodiment 1
[0024] Preparation of fermentation medium: glucose 20g / L, peptone 2g / L, potassium dihydrogen phosphate 0.5g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate 0.2g / L, calcium chloride 0.2g / L, pantothenic acid 0.01g / L, vitamin B1 0.05g / L, pH 6, sterilized at 121°C for 15 minutes, cooled to about 30°C for later use.
[0025] 30L of culture medium was loaded into a 50L stirring tank, and after sterilization, the Penicillium citrinum strain cultured on the wort slant medium was inserted for fermentation and cultivation. Samples were taken at regular intervals to measure the enzyme activity of nuclease P1. After culturing for 56 hours, the enzyme activity stabilized at about 3300U / mL. 24L of fermentation broth was released, and 6L was kept in the tank as the seed solution for the next batch of fermentation. Prepare 24L fermentation medium and put it into the fermenter for further cultivation. After 22 hours, the enzyme activity was stabilized to 4800U / mL, and 24L of ferme...
Embodiment 2
[0028] Preparation of fermentation medium: glucose 30g / L, peptone 3g / L, potassium dihydrogen phosphate 1g / L, dipotassium hydrogen phosphate 1g / L, magnesium sulfate 0.5g / L, calcium chloride 0.2g / L, biotin 0.1 g / L, niacin 0.06g / L, vitamin B2 0.2g / L, pH 7, sterilized at 121°C for 15min, cooled to about 30°C for later use.
[0029] A 1.2T culture medium was placed in a 2T stirring tank, and after sterilization, the Penicillium citrinum strain cultured on a wort slant medium was inserted for fermentation. Samples were taken at regular intervals to measure the enzyme activity of nuclease P1. After 58 hours of cultivation, the enzyme activity stabilized at about 3700U / mL, and 1T of fermentation broth was released, and 0.2T was kept in the tank as the seed solution for the next batch of fermentation. Prepare 1T fermentation medium and put it into the fermenter for further cultivation. After 20 hours, the enzyme activity stabilized to 5200U / mL, released 1T fermentation broth again, an...
Embodiment 3
[0032] Preparation of fermentation medium: glucose 40g / L, peptone 4g / L, potassium dihydrogen phosphate 1g / L, dipotassium hydrogen phosphate 1g / L, magnesium sulfate 0.5g / L, calcium chloride 0.2g / L, pantothenic acid 0.1g / L, niacin 0.2g / L, folic acid 0.5g / L, pH 7, sterilized at 121°C for 15min, cooled to about 30°C for later use.
[0033] A 12T culture medium was put into a 20T stirring tank, and after sterilization, the Penicillium citrinum strain cultured on the wort slant medium was inserted for fermentation. Samples were taken at regular intervals to measure the enzyme activity of nuclease P1. After culturing for 56 hours, the enzyme activity stabilized at about 4500U / mL, 8T of fermentation broth was released, and 4T was kept in the tank as the seed solution for the next batch of fermentation. Prepare 8T fermentation medium and put it into the fermenter for further cultivation. After 20 hours, the enzyme activity stabilized to 5400U / mL, and the 8T fermentation broth was rel...
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