Saccharomyces cerevisiae strain capable of rapidly assimilating ammonia and application thereof
By screening and applying Saccharomyces cerevisiae HBX01 to prepare microbial preparations, the problem of removing ammonia and hydrogen sulfide from livestock and poultry manure was solved, achieving efficient and stable deodorization and reducing the concentration of harmful gases in the manure.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- JIANGNAN UNIV
- Filing Date
- 2022-11-28
- Publication Date
- 2026-05-29
AI Technical Summary
The lack of efficient and environmentally adaptable single microbial strains in existing technologies for the removal of ammonia and hydrogen sulfide from livestock and poultry manure leads to unstable effects of existing microbial deodorizers, making it difficult to achieve efficient deodorization.
A rapidly ammonia-assimilating Saccharomyces cerevisiae hbx01 strain is provided, which is used to prepare microbial preparations containing Saccharomyces cerevisiae, including live cell cultures or solid preparations, for spraying in livestock and poultry manure to adsorb and assimilate ammonia and hydrogen sulfide.
This brewing yeast can significantly reduce the concentration of ammonia and hydrogen sulfide in livestock and poultry manure. Within 24 hours, ammonia levels dropped from 16.6 mg/m3 to 8.2 mg/m3, and within 48 hours, they dropped to 4.8 mg/m3. Hydrogen sulfide levels dropped from 0.26 mg/m3 to 0.04 mg/m3, which is more effective than commercially available deodorizers.
Abstract
Description
Technical Field
[0001] This invention relates to a rapidly ammonia-assimilating brewing yeast and its applications, belonging to the field of microbial technology. Background Technology
[0002] Animal husbandry is an important part of agriculture. With the continuous improvement of living standards in my country, the demand for meat, eggs, and dairy products has further increased, promoting the large-scale and intensive development of my country's animal husbandry. However, odor pollution caused by livestock and poultry manure has become a prominent environmental pollution problem in my country. The odor from manure is mostly composed of harmful gaseous substances, mainly including ammonia and hydrogen sulfide, which harm the respiratory system of livestock and poultry, reduce their resistance, induce various infectious diseases, and affect their healthy growth. At the same time, the unregulated emission of odorous gases also pollutes the surrounding environment and affects the health of residents. According to data from the "China Ambient Air Quality Management Assessment Report," my country's agricultural NH3 emissions are approximately 10 million tons, of which 90% are from agriculture and livestock and poultry farming. Based on the NH3 emission inventory of major livestock and poultry farming provinces, livestock and poultry sources account for approximately 75% of the total NH3 emissions.
[0003] There are numerous technologies for treating fecal odor both domestically and internationally, primarily including physical, chemical, biological, or combinations of these methods. The principle behind all these odor control technologies is to alter the phase or structure of odorous substances or reduce their intensity through physical, chemical, or biological processes, thereby achieving the goal of removing or reducing the odor. Physical methods mainly rely on the transformation between solid, liquid, and gas phases to eliminate odors and reduce the degree of olfactory perception. However, the chemical properties are not fundamentally changed, and under certain conditions, the odor can still be released as a gaseous substance. These methods are only suitable for treating low-concentration, small-scale odors, and are expensive, with the risk of secondary pollution if not handled properly. Chemical deodorization technology mainly uses chemical reagents to react with odorous substances, altering their chemical structure and transforming them into odorless or less odorous substances. A well-developed chemical deodorization method has high efficiency, capable of completely oxidizing and decomposing odorous substances; however, it requires numerous and complex equipment, consumes a lot of energy, is costly, and has a short duration of effect. Biological deodorization has advantages such as low operating and maintenance costs, high deodorization efficiency, convenient operation, and good safety, and has attracted the attention of researchers at home and abroad.
[0004] Livestock and poultry manure is a complex micro-ecosystem with diverse indigenous microbial communities. Screening for highly efficient microorganisms that can inhibit the production of ammonia and hydrogen sulfide and assimilate existing ammonia, and then allowing them to thrive dominantly in the manure, is crucial for controlling biological odors. However, due to the lack of screening methods for single, highly efficient deodorizing microbial strains, current methods often result in single microorganisms with weak environmental adaptability, making it difficult for them to become the dominant flora in odor-emitting sources and achieve effective deodorization. Therefore, current microbial deodorizers are mostly mixed strain formulations, and the deodorizing efficacy of each strain component in the composition is not disclosed. However, due to the diversity of microbial composition, metabolic types, respiratory types, and functions, this complex microbial metabolic system often cannot be precisely controlled, leading to inconsistent and unreliable deodorization effectiveness. Summary of the Invention
[0005] The technical problem to be solved by the present invention is to provide a yeast strain that rapidly assimilates ammonia and its application in the removal of ammonia and hydrogen sulfide in farms.
[0006] This invention provides a strain of Saccharomyces cerevisiae hbx01, which was deposited at the China Center for Type Culture Collection on October 19, 2022, with accession number CCTCC NO:M 20221597.
[0007] The present invention also provides a microbial preparation containing the aforementioned brewing yeast.
[0008] In one embodiment, the microbial preparation includes, but is not limited to, a cell culture containing live cells of the Saccharomyces cerevisiae hbx01.
[0009] In one embodiment, the microbial preparation is a diluted cell culture containing live cells of the brewer's yeast hbx01.
[0010] In one embodiment, the microbial preparation is prepared by mixing the cell cells of the brewing yeast hbx01 with sorbitan monostearate and potato starch, followed by fluidized bed drying and granulation to obtain a solid microbial preparation.
[0011] The present invention also provides the application of the brewing yeast in the removal of ammonia and / or hydrogen sulfide gas.
[0012] In one embodiment, the application is used to reduce the content of ammonia and / or hydrogen sulfide in a livestock farm.
[0013] In one embodiment, the application is to reduce the levels of ammonia and / or hydrogen sulfide in the manure-containing environment of a farm.
[0014] In one embodiment, the application involves spraying a liquid containing the brewing yeast onto the surface of the material to be treated.
[0015] In one embodiment, the liquid containing brewer's yeast is prepared by inoculating the brewer's yeast into a 5brix wort culture medium, fermenting at 20-30°C for at least 18 hours, and then diluting it with water 20-30 times.
[0016] In one embodiment, the material to be processed includes, but is not limited to, feces.
[0017] In one embodiment, the application involves spraying the brewing yeast onto the surface of the material at an amount not less than 0.25% of the material's wet weight.
[0018] In one embodiment, the application involves spraying the liquid containing the brewer's yeast onto the feces, once every 6 to 8 hours, with each spray amount being 0.35% of the wet weight of the feces, for a treatment period of 24 to 48 hours.
[0019] In one embodiment, the application involves spraying the liquid containing the brewer's yeast onto the feces, once every 6 hours, with each spray amount being 0.25% of the wet weight of the feces, for a treatment period of 24–48 hours.
[0020] In one embodiment, the application involves spraying the liquid containing the brewer's yeast onto the feces, once every 8 hours, with each spray amount being 0.35% of the wet weight of the feces, for a treatment period of 24–48 hours.
[0021] Beneficial effects:
[0022] This invention provides a brewing yeast strain capable of rapidly adsorbing and assimilating ammonia and hydrogen sulfide gases. This yeast produces glutamate dehydrogenase with high activity, capable of ammonifying α-ketoglutarate formed during its own metabolism to produce glutamate. The glutamate dehydrogenase activity is 38.6 U / mL, and it can assimilate NH4+ in 12 hours. + 0.8 g / L, can assimilate NH4 in 18 hours + 1.9g / L.
[0023] This invention utilizes the brewing yeast in the treatment of ammonia and hydrogen sulfide; after 24 hours of treatment, the ammonia level in the environment can be reduced from 16.6 mg / m³. 3 Reduced to 8.2 mg / m³ 3 Hydrogen sulfide from 0.26 mg / m³ 3 Reduced to 0.08 mg / m³ 3 After 48 hours, the ammonia level in the environment decreased to 4.8 mg / m³. 3 Hydrogen sulfide decreased to 0.04 mg / m³. 3 .
[0024] Preservation of biological materials
[0025] Saccharomyces cerevisiae hbx01, classified as Saccharomyces cerevisiae hbx01, was deposited on October 19, 2022, at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC NO:M 20221597, located at Wuhan University, Wuhan, China. Detailed Implementation
[0026] Methods for detecting ammonia: Use a two-in-one gas detector to measure the concentration of ammonia and hydrogen sulfide in the environment.
[0027] Methods for detecting hydrogen sulfide content: Use a two-in-one gas detector to measure the concentration of ammonia and hydrogen sulfide in the environment.
[0028] Example 1: Screening of brewing yeast
[0029] (1) Add 2.5% alcohol to YPD medium to enrich and isolate yeast from various farmed animal manure and fermented food;
[0030] (2) Using 5Brix wort as the culture medium, the yeast isolated in step (1) was inoculated. The initial screening was performed when the pH dropped to 4.5 or below within 24 hours. The yeast obtained from the initial screening was subjected to glutamate dehydrogenase activity determination, and yeast with high glutamate dehydrogenase activity was obtained by secondary screening. Determination of yeast glutamate dehydrogenase: Take 2 mL of yeast culture in a centrifuge tube, centrifuge at 10,000×g for 10 min (4℃), discard the supernatant, add 1 mL of extraction solution, and sonicate the yeast (ice bath, power 200W, sonication for 3s, interval 10s, repeated 30 times), centrifuge at 8,000×g for 10 min (4℃), take the supernatant, and determine the enzyme activity using a glutamate dehydrogenase detection kit.
[0031] (3) For the yeast strains obtained from the secondary screening, a 24-hour fermentation experiment was conducted by adding 5 g / L glucose to the yeast nitrogen source basal medium (YNB medium). The ammonia content was measured at 12 h, 18 h, and after fermentation to identify the target yeast, which was then identified and preserved. The results showed that *Saccharomyces cerevisiae* hbx01 could assimilate NH4+ after 12 h of culture. + 0.8 g / L, can assimilate NH4 in 18 hours + 1.9g / L.
[0032] Example 2: Production of glutamate dehydrogenase by Saccharomyces cerevisiae
[0033] Using 5 Brix wort as the culture medium, the *Saccharomyces cerevisiae* hbx01 obtained from screening in Example 1 was inoculated and fermented at 30°C for 18 hours to obtain a bacterial concentration of 5 × 10⁻⁶.7 Yeast culture medium with CFU / mL.
[0034] Determination of yeast glutamate dehydrogenase: Take 2 mL of yeast culture in a centrifuge tube, centrifuge at 10,000 × g for 10 min (4℃), discard the supernatant, add 1 mL of the detection kit extraction solution (containing 0.1 M Tris-HCl at pH 8.3), sonicate to disrupt the yeast (ice bath, power 200 W, sonication for 3 s, interval 10 s, repeated 30 times), centrifuge at 8,000 × g for 10 min (4℃), take the supernatant, and determine the enzyme activity using the glutamate dehydrogenase detection kit. The glutamate dehydrogenase activity was 38.6 U / mL.
[0035] Example 3: Application of Saccharomyces cerevisiae in ammonia assimilation
[0036] Using 5 Brix wort as the culture medium, the *Saccharomyces cerevisiae* hbx01 obtained from screening in Example 1 was inoculated at 5% (v / v) and fermented at 30°C for 18 hours, yielding a bacterial concentration of 5 × 10⁻⁶. 7 Yeast culture medium at CFU / mL was diluted 10 times with water and incubated at 30℃ for 6 hours. The solution was then sprayed onto manure in a closed indoor environment of the farm, once every 6 hours. Each spray was 0.25% of the wet weight of the manure. A control group without spraying was used. Within 24 hours of spraying the yeast culture medium, the ammonia level in the environment increased from 16.6 mg / mL. 3 Reduced to 8.2 mg / m³ 3 Hydrogen sulfide from 0.26 mg / m³ 3 Reduced to 0.08 mg / m³ 3 After 48 hours, the ammonia level in the environment decreased to 4.8 mg / m³. 3 Hydrogen sulfide decreased to 0.04 mg / m³. 3 .
[0037] Example 4: Application of Saccharomyces cerevisiae HBX01 in ammonia assimilation
[0038] Using 5 Brix wort as the culture medium, the *Saccharomyces cerevisiae* hbx01 obtained from screening in Example 1 was inoculated at 5% (v / v) into the 5 Brix wort medium and fermented at 30°C for 18 hours to obtain a bacterial concentration of 5 × 10⁻⁶. 7 Yeast culture medium at CFU / mL. The fermentation broth was diluted 30 times with water and then sprayed onto the feces every 8 hours, with each spray amount being 0.35% of the wet weight of the feces. Within 24 hours, the ammonia level in the environment increased from 16.7 mg / mL. 3 Reduced to 12.4 mg / m³ 3 Hydrogen sulfide from 0.28 mg / m³ 3 Reduced to 0.12 mg / m³ 3After 48 hours, the ammonia level in the environment decreased to 9.8 mg / m³. 3 Hydrogen sulfide decreased to 0.08 mg / m³. 3 .
[0039] Example 5: Preparation of Saccharomyces cerevisiae HBX01 microbial preparation
[0040] Using 5Brix wort as a culture medium, the brewing yeast hbx01 obtained in Example 1 was inoculated and fermented at 30°C for 18-24 hours to obtain a yeast culture broth, which can be used directly or diluted as a liquid preparation of brewing yeast hbx01.
[0041] The yeast culture was centrifuged to collect the cells. An appropriate amount of sorbitan monostearate and potato starch were added, and the mixture was dried and granulated using a fluidized bed to obtain a solid formulation containing Saccharomyces cerevisiae hbx01.
[0042] Comparative Example 1
[0043] Using wort without brewer's yeast as a control, the specific steps were as follows: 5 Brix wort was diluted 10 times with water and then sprayed onto the manure every 6 hours, with each spray amount being 0.25% of the wet weight of the manure. Within 24 hours, the ammonia level in the environment increased from 15.8 mg / m³. 3 Rising to 16.4 mg / m³ 3 Hydrogen sulfide from 0.24 mg / m³ 3 Reduced to 0.22 mg / m³ 3 After 48 hours, the ammonia level in the environment decreased to 14.8 mg / m³. 3 Hydrogen sulfide decreased to 0.20 mg / m³. 3 .
[0044] Comparative Example 2
[0045] The specific implementation method is the same as in Example 3, except that the brewer's yeast is replaced with brewer's yeast S288C. The brewer's yeast is cultivated according to the method in Example 2. The yeast culture solution is diluted 10 times with water and kept at 30°C for 6 hours. Then it is sprayed on the manure in the closed indoor environment of the farm, once every 6 hours. The amount sprayed each time is 0.25% of the wet weight of the manure. The ammonia level in the environment is 11.6 mg / m³ within 24 hours. 3 Hydrogen sulfide 0.16 mg / m³ 3 After 48 hours, the ammonia level in the environment was 10.8 mg / m³. 3 Hydrogen sulfide 0.12 mg / m³ 3 .
[0046] The applicant also attempted to compare the deodorizing effect with that of commercially available deodorizers. The results showed that the Saccharomyces HBX01 screened in this invention had a significantly better effect on reducing ammonia and hydrogen sulfide than commercially available deodorizers.
[0047] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.
Claims
1. A strain of brewer's yeast ( Saccharomyces cerevisiae hbx01 was deposited at the China Center for Type Culture Collection on October 19, 2022, with accession number CCTCC NO: M 20221597.
2. A microbial preparation containing the brewing yeast of claim 1.
3. The microbial preparation according to claim 2, characterized in that, The microbial preparation is a cell culture containing live cells of the brewer's yeast hbx01.
4. The microbial preparation according to claim 2, characterized in that, The microbial preparation is a solid microbial preparation obtained by mixing the cell cells of the brewing yeast hbx01 with sorbitan monostearate and potato starch, followed by fluidized bed drying and granulation.
5. A method for reducing the content of ammonia and hydrogen sulfide in the environment, characterized in that, The brewing yeast hbx01 of claim 1 is used to treat substances in the environment that produce ammonia and hydrogen sulfide; the environment is a farm environment containing manure; the treatment involves spraying a liquid containing the brewing yeast hbx01 of claim 1 onto the surface of the material to be treated.
6. The method according to claim 5, characterized in that, The liquid is prepared by inoculating the brewing yeast hbx01 into a wort culture medium, fermenting it at 20-30°C for at least 18 hours, and then diluting it with water 20-30 times.
7. The method according to claim 5 or 6, characterized in that, Spray the brewing yeast hbx01 described in claim 1 onto the surface of the material at an amount of not less than 0.25% of the wet weight of the material, and treat for at least 24 hours.
8. The application of the brewing yeast hbx01 according to claim 1, or the microbial preparation according to any one of claims 2 to 4, or the method according to any one of claims 5 to 7 in the removal of ammonia and hydrogen sulfide gases.