Preparation of nitrite reductase and method for preparing nitrite reductase preparation
A technology of reductase and nitrous acid, which is applied in the field of preparation of biological enzymes and enzyme preparations, achieves the effects of short production cycle, reduction of human diseases, and promotion of quality improvement
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Embodiment 1
[0030] 1. Culture Medium Preparation
[0031] (1) Formula of slant seed culture medium: 1-2wt% of nutrient broth powder, 1.8-2wt% of agar, pH6.5-7.5.
[0032] (2) Fermentation medium formula: glucose 1-2wt%, peptone 0.1wt%, sodium chloride 0.1-0.5wt%, dipotassium hydrogen phosphate 0.1-0.3wt%, potassium dihydrogen phosphate 0.05-0.2wt%, MgSO 4 ·7H 2 O 0.02-0.06wt%, MnSO 4 ·H 2 O 0.001-0.003wt%, CaCl 2 2H 2 O 0.002-0.004wt%, pH6.5-7.5.
[0033] 2. Dissolve the lyophilized powder of Bacillus megaterium in physiological saline at a weight ratio of 1:4, pick a ring with an inoculation loop, inoculate it on the slant seed medium, and cultivate it at 37°C for 20 hours to obtain an activated slant; ferment with 10mL The culture medium washes the activated slant bacteria under sterile conditions, puts it into a 250mL shake flask, fills each bottle with 100mL seed medium, and inoculates 2-4 seeds on each slant, at 25-30°C, with a rotation speed of 100-240r / min Cultivate at a con...
Embodiment 2
[0042] 1. Obtain the cell preparation
[0043] (1) Formula of slant seed medium: glucose 15.0g, NaNO 2 1.0g, K 2 HPO 4 1.35g, KH 2 PO 4 0.70g, MgSO 4 ·7H 2 O 0.20g, agar 20.0g, dilute to 1L, adjust the pH to 7.0, and sterilize at 121°C for 20min.
[0044] (2). Fermentation medium formula: glucose 15.0, peptone 2.0g, Na 2 HPO 4 1.35g, KH 2 PO 4 0.70g, MgSO 4 ·7H 2 O 0.10g, NaCl 1.0g, CaCl 2 2H 2 O 0.02g, MnSO 4 ·H 2 O 0.01g, NaNO 2 , 0.138g, dilute to 1L, adjust the pH to 7.0, and sterilize at 121°C for 20min.
[0045] 2. Dissolve the lyophilized powder of Bacillus megaterium in physiological saline at a weight ratio of 1:2, pick a ring with an inoculation loop, inoculate it on the slant medium, and culture it at 37° C. for 20 hours to obtain an activated slant. Then use the inoculation loop to pick 3-4 rings and inoculate them in 250mL shake flasks, each bottle contains 50mL of seed culture medium, and culture at 30-35°C for 24 hours at a constant tempera...
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