Organic phosphorus degrading enzyme preparation and preparation method thereof
An organophosphorus and enzyme-degrading technology, applied in the field of microorganisms, can solve problems such as toxicity and achieve better results
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Embodiment 1
[0013] 1) Fermentation of seed liquid: Beef extract peptone medium was sterilized and inoculated with Methylobacterium at an inoculum size of 10% by volume, and cultured at 30° C. and 180 r / min for 60 h.
[0014] 2) Fermentation tank fermentation: move the seed liquid into the seed tank according to the inoculation amount of 8% by volume, and carry out fermentation and cultivation. The speed is 300 rpm, and the fermentation time is 60 hours.
[0015] 3) Centrifuge the fermentation broth at 4°C, 7000r / min. for 30min, discard the supernatant, and add 1 / 40 of the fermentation broth volume pH7.5, 0.05mol / L of EDTA and 1 / 15 of the fermentation broth volume to the bacteria sludge Gently shake the sucrose solution to make the bacteria fully suspend, put it in ice bath for 60min, centrifuge at 7000r / min for 30min at 4°C, discard the hypertonic solution, add 1 / 40 of the volume of the fermentation broth to the sludge, and gently Shake well, put in an ice bath for 60 minutes, centrifuge...
Embodiment 2
[0023] The difference from Example 1 is:
[0024] 1) Seed liquid fermentation: Beef extract peptone medium was sterilized and inoculated with Methylobacterium at an inoculum size of 5% by volume, cultured at 35° C. and 180 r / min for 48 hours.
[0025] 2) Fermentation tank fermentation: move the seed liquid into the seed tank according to the inoculation amount of 5% by volume, and carry out fermentation and cultivation. The conditions are: tank temperature 25°C, tank pressure 0.02MPa, ventilation rate 1:1.0v / v / min, stirring The speed is 300 rpm, and the fermentation time is 48 hours.
[0026] 3) Centrifuge the fermentation broth at 4°C, 6000r / min for 40min. Discard the supernatant, add 1 / 30 of the fermentation broth volume pH7.5, 0.05mol / L of EDTA and 1 / 10 of the fermentation broth volume to the sludge Gently shake the sucrose solution to make the bacteria fully suspended, ice bath for 30min, centrifuge at 4°C, 6000r / min for 40min, discard the hypertonic solution, add 1 / 30 of...
Embodiment 3
[0034] The difference from Example 1 is:
[0035] 1) Fermentation of seed liquid: Beef extract peptone medium was sterilized and inoculated with Methylobacterium at an inoculum size of 5% by volume, cultured at 32° C. and 180 r / min for 52 hours.
[0036] 2) Fermentation tank fermentation: transfer the seed liquid into the seed tank according to the inoculation amount of 5% by volume, and carry out fermentation and cultivation. The speed is 300 rpm, and the fermentation time is 52 hours.
[0037] 3) Centrifuge the fermentation broth at 4°C, 8000r / min for 35min. Discard the supernatant, add 1 / 30 of the fermentation broth volume pH7.5, 0.05mol / L of EDTA and 1 / 10 of the fermentation broth volume to the sludge Gently shake the sucrose solution to make the bacteria fully suspended, ice bath for 50min, centrifuge at 8000r / min for 35min at 4°C, discard the hypertonic solution, add 1 / 30 of the fermentation liquid volume of ice water to the bacterial sludge, and gently Shake well, put...
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