A kind of preparation method and application of cytochrome p450 enzyme in bacteria
A cytochrome and bacterial technology, applied in biochemical equipment and methods, chemical instruments and methods, enzymes, etc., can solve the problem of less antibiotics and achieve high extraction rate, high purity, and good activity
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Embodiment 1
[0026] Bacteria culture: Bacillus B-1 was inoculated in the bacterial culture solution with an inoculation amount of 3wt%, and cultured for 16 days in an environment with a temperature of 20°C, a salinity of 30‰, and a pH value of 8 to obtain a bacteria solution . Among them, the composition of the bacterial culture solution is: sulfadiazine 1mg / L, sodium acetate 2g / L, sodium nitrate 0.1g / L, ammonium chloride 0.1g / L, dipotassium hydrogen phosphate 0.05g / L, yeast extract powder 0.05 g / L, the solvent is sea water.
[0027] Freezing-crushing treatment: use a refrigerated centrifuge to centrifuge at a constant temperature of 4°C at a speed of 15,000r / min for 5 minutes, remove the supernatant, wash the obtained bacteria with sterile artificial seawater, and centrifuge twice to remove the medium. The centrifuge tube was frozen at -18°C for 4.5 hours, then taken out and controlled in an ice-water bath with an ultrasonic breaker at 20KHz (10s / 10s) for 15min (power 1000W, temperature ...
Embodiment 2
[0031] Bacteria culture: Bacillus B-1 was inoculated in the bacterial culture solution with an inoculation amount of 4wt%, and cultured for 10 days in an environment with a temperature of 25°C, a salinity of 20‰, and a pH value of 7.5 to obtain a bacteria solution . Among them, the composition of the bacterial culture solution is: sulfadiazine 1mg / L, sodium acetate 2g / L, sodium nitrate 0.1g / L, ammonium chloride 0.1g / L, dipotassium hydrogen phosphate 0.05g / L, yeast extract powder 0.05 g / L, the solvent is sea water.
[0032] Freezing-crushing treatment: use a refrigerated centrifuge to centrifuge at a constant temperature at 4°C, centrifuge at a speed of 15,000r / min, centrifuge for 5min, remove the supernatant, wash the obtained bacteria with sterile artificial seawater and centrifuge again for 3 times, remove the medium, and centrifuge The tube was frozen at -15°C for 5 hours, then taken out and controlled in an ice-water bath and crushed with an ultrasonic breaker at 25KHZ (1...
Embodiment 3
[0036] Bacteria culture: Bacillus B-1 was inoculated in the bacterial culture solution with an inoculation amount of 5wt%, and cultivated for 8 days in an environment with a temperature of 30°C, a salinity of 35‰, and a pH value of 7 to obtain a bacteria solution . Among them, the composition of the bacterial culture solution is: sulfadiazine 1mg / L, sodium acetate 2g / L, sodium nitrate 0.1g / L, ammonium chloride 0.1g / L, dipotassium hydrogen phosphate 0.05g / L, yeast extract powder 0.05 g / L, the solvent is sea water.
[0037] Freezing-crushing treatment: use a refrigerated centrifuge to centrifuge at a constant temperature at 4°C, centrifuge at a speed of 15,000r / min, centrifuge for 5min, remove the supernatant, wash the obtained bacteria with sterile artificial seawater and centrifuge again for 3 times, remove the medium, and centrifuge The tube was frozen at -20°C for 4 hours, then taken out and controlled in an ice-water bath and crushed with an ultrasonic breaker at 22KHz (10...
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