Pichia farinosa NHG and application of Pichia farinosa to in degradation of ammonia nitrogen
A technology of powdered Pichia pastoris and ammonia nitrogen degradation, applied in the field of microorganisms, can solve the problems of secondary pollution, high operating cost of ammonia nitrogen and ammonia gas discharge, and achieve the effects of high safety, improving the environment in the house, and increasing the utilization rate
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Embodiment 1
[0028] Example 1. Separation, purification and identification of powdery Pichia pastoris NHG
[0029] 1. Separation and purification of powdered Pichia pastoris NHG
[0030] 1. Enrichment of ammonia nitrogen degrading bacteria
[0031] Take 20 g of chicken manure samples fermented for 3 days, inoculate them in a conical flask filled with 100 ml of sterilized distilled water (add sterilized glass beads to the bottle), vibrate at 30°C and 150 r / min for 30 min to fully break up the bacteria micelles, and statically Set for 20min. Take 5ml of the supernatant, inoculate it into a conical flask containing 100ml of enrichment medium, culture it with shaking at 30°C and 150r / min for 2 days, take 1ml of the culture medium and inoculate it again into a new enrichment medium, and repeat continuously Enriched for 10 generations.
[0032] 2. Screening, isolation and purification of ammonia nitrogen degrading bacteria
[0033] Draw 1ml of the enriched culture solution, and after 10 -1 ...
Embodiment 2
[0044] Example 2, research on the function of powdery Pichia pastoris NHG to degrade ammonia nitrogen
[0045] 1. Research on the degradation characteristics of Pichia pastoris NHG to high concentration ammonia nitrogen
[0046] 1. Preparation of Medium A
[0047] Medium A: Glucose was used as the sole carbon source, and the carbon source and (NH 4 ) 2 S0 4 The concentration of ammonia nitrogen is 300mg / L, C / N is 10, and pH is 7.0.
[0048] 2. Pick a single clone of Pichia powder pastoris NHG, inoculate it in 100mL YPD liquid medium, and culture it with shaking at 30°C and 150r / min for 24-36h (to make the cell concentration in the culture system reach 8.0-9.0×10 8 cfu / mL), centrifuged at 4000rpm for 10min, collected the bacteria, and washed 3 times with sterile saline.
[0049] 3. Take the bacterial cells obtained in step 2 and resuspend them with sterile physiological saline to obtain a bacterial concentration of 8.0×10 8 cfu / mL bacterial suspension.
[0050] 4. Inocul...
Embodiment 3
[0075] Embodiment 3, the influence of powdered Pichia pastoris NHG on the amount of excrement ammonia emission
[0076] 1. Pick a single clone of Pichia powdery NHG, inoculate it in 100mL YPD liquid medium, and culture it with shaking at 30°C and 150r / min for 24-36h (to make the cell concentration in the culture system reach 8.0-9.0×10 8 cfu / mL), centrifuged at 4000rpm for 10min, collected the bacteria, and washed 3 times with sterile saline.
[0077] 2. Take the bacterial cells obtained in step 1 and resuspend them with sterile physiological saline to obtain a bacterial concentration of 8.0×10 8 cfu / mL bacterial suspension.
[0078]3. Take a large beaker (1L), put a small beaker in the large beaker (the small beaker is equipped with 20ml of absorption liquid; the absorption liquid is 0.01mol / L sulfuric acid solution), put 100g of fresh chicken manure in the large beaker The sample and 10ml of the bacterial suspension obtained in step 2 (using an equal volume of sterile sali...
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