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Pichia powder pastoris nhg and its application in degrading ammonia nitrogen

A powdered Pichia pastoris, ammonia nitrogen degradation technology, applied in the field of microorganisms, can solve the problems of high ammonia nitrogen and ammonia gas emission, secondary pollution and other problems of operating costs, and achieve the effects of improving animal health and welfare and protecting human ecological environment.

Inactive Publication Date: 2018-05-01
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the existing measures to control ammonia pollution in farms use physical methods (such as strengthening the ventilation in the house, building excrement and urine separation facilities, etc.), the operating cost is high, and because the discharge of ammonia nitrogen and ammonia to the surrounding environment has not been fundamentally eliminated, it is still a problem. It will cause secondary pollution, and the use of microbial methods can overcome the defects of the above methods, so it has a high application value in the treatment of ammonia and ammonia nitrogen pollution in animal husbandry

Method used

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  • Pichia powder pastoris nhg and its application in degrading ammonia nitrogen
  • Pichia powder pastoris nhg and its application in degrading ammonia nitrogen
  • Pichia powder pastoris nhg and its application in degrading ammonia nitrogen

Examples

Experimental program
Comparison scheme
Effect test

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 Erlenmeyer 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 ...

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 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.

[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), and 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 ...

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Abstract

The invention discloses Pichia farinosa NHG and application of the Pichia farinosa to in the degradation of ammonia nitrogen. The preservation registration number of the Pichia farinosa NHG provided by the invention is CGMCC (China General Microbiological Culture Collection Center) NO.10561. The invention also protects a culture and a bacterial suspension of the Pichia farinosa NHG. The invention further protects a product; active ingredients of the product are the Pichia farinosa NHG, the culture or the bacterial suspension; the functions of the product are described as (a) or, (b) or, (c) or (d): (a) degrading the ammonia nitrogen; (b) degrading ammonia nitrogen in water; (c) degrading ammonia nitrogen in excrement; (d) reducing the emission of ammonia gas in the excrement, wherein the excrement can be livestock excrement, such as chicken manure. The Pichia farinosa provided by the invention can be applied to the following aspects of (1) improving the water quality problem of water polluted by the ammonia nitrogen, (2) improving the environmental sanitation in livestock and poultry houses, and (3) being used as a feed additive of chicken.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of Pichia powder and the application of the Pichia powder in degrading ammonia nitrogen. Background technique [0002] The waste water and feces produced in the process of livestock and poultry breeding will produce a large amount of malodorous gas during the storage process, among which the harm of ammonia gas is the most obvious. The lighter ones induce the occurrence of respiratory diseases in poultry, pigs and other animals, causing animal stress reactions, and the severe ones Those who can cause poisoning death. At the same time, a large amount of ammonia emission promotes the formation of particulate matter (PM2.5) that can enter the lungs, and diffuses into the atmosphere to react with some acidic substances to form ammonium salt aerosols, leading to eutrophication and acidification of the ecosystem. In addition, if the livestock and poultry br...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16B01D53/84B01D53/58C02F3/34C12R1/84C02F101/16
CPCY02A50/20
Inventor 刘国华刘志云司彦培石鹏君蔡辉益常文环张姝郑爱娟
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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