CN111893054B - Bacillus subtilis KY07 and application thereof in deodorization - Google Patents
Bacillus subtilis KY07 and application thereof in deodorization Download PDFInfo
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Abstract
The invention relates to a Bacillus subtilis KY07 and application thereof in deodorization, wherein the Bacillus subtilis KY07 is classified and named and is preserved in China center for type culture collection with the preservation number of CCTCC NO: M2020152. The strain can effectively degrade ammonia nitrogen and hydrogen sulfide, can effectively degrade odor generated by fermentation of kitchen garbage when used for treating the kitchen garbage, reduces odor pollution, and has the advantages of high efficiency and no need of additional energy consumption. The method for treating kitchen garbage odor can obviously reduce energy consumption in the process of recycling kitchen garbage, save energy and reduce emission, and bring more economic value for recycling kitchen garbage.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a bacillus subtilis KY07 and application thereof in deodorization.
Background
The aerobic fermentation resource utilization of the kitchen waste is one of novel treatment modes for treating the kitchen waste at present, however, a large amount of ammonia gas and hydrogen sulfide can be generated in the aerobic fermentation process, and both the ammonia gas and the hydrogen sulfide have pungent smell and corrosivity, so that the human health is injured, and the environment is polluted. Ammonia gas and hydrogen sulfide are the most main components of the fermentation odor of the kitchen waste.
The ammonia gas is very easy to dissolve in water and reacts with acid to generate ammonium salt, the ammonium salt exists mainly in two forms of ammonium sulfate and ammonium nitrate, ammonium nitrogen is one of the main existing forms of the ammonia gas, the ammonium nitrogen is easily changed into the ammonia gas to escape from a water environment under an alkaline condition, the ammonia can stimulate the mucous membrane of the upper respiratory tract of a human, and a serious person can poison the human and threaten the life of the human.
The hydrogen sulfide is a nerve poison, has the odor of smelly eggs, is a main component of malodorous gas, is one of air pollutants with high toxicity, not only brings stimulation and dislike to the sense of people, but also directly harms the body health of people. The low-concentration hydrogen sulfide can generate obvious stimulation effects, such as lacrimation, ophthalmalgia and the like to cause conjunctivitis, and is very easy to be absorbed by intestines, stomachs and lungs to cause chest pain, chest distress, nausea and the like, and dyspnea, palpitation, syncope and the like can be caused in severe cases; high concentration hydrogen sulfide can paralyze the olfactory nerve ending of a human, so that the human cannot smell the odor of the hydrogen sulfide, thereby causing the human to be paralyzed, syncope and even die.
The ammonia gas and hydrogen sulfide generated by the aerobic fermentation of the kitchen waste pollute the environment and bring serious influence to the physical and mental health of surrounding residents. Therefore, how to remove ammonia and hydrogen sulfide efficiently and at low cost is one of the technical problems of aerobic fermentation and resource utilization of kitchen waste, and although some commercial biomass waste treatment equipment is provided with a deodorization system, the problems of poor deodorization effect, high energy consumption, high cost and the like still exist.
Disclosure of Invention
The invention aims to provide a bacillus subtilis KY07 and application thereof in deodorization, wherein the bacillus subtilis KY07 can effectively degrade ammoniacal nitrogen and hydrogen sulfide in the fermentation process of kitchen garbage, and effectively reduce odor pollution caused by fermentation of the kitchen garbage.
Therefore, the first aspect of the invention provides a Bacillus subtilis KY07 which is classified and named as Bacillus subtilis KY07 and is preserved in China center for type culture Collection (CCTCC for short), wherein the preservation unit address is Wuhan university No. 299 in Wuchang district, Wuhan city, Hubei, China center for type culture Collection, the preservation number is CCTCC NO: M2020152, and the preservation date is 2020, 5 and 28 days.
The bacillus subtilis KY07 has the following properties: after culturing for 48h at 37 ℃ in an LB solid plate culture medium, colonies are light yellow, opaque, neat in edges, wrinkled in the middle, sporulated, positive in gram stain, and rod-shaped thalli are observed under a microscope.
DNA sequencing is carried out on the Bacillus subtilis KY07, a sequencing result is compared with a corresponding database, and sequence determination and sequence comparison are carried out on a http:// https:// blast.ncbi.nlm.nih.gov/blast.cgi website, so that the highest sequence homology of the gene sequence of the strain and the registered Bacillus subtilis (Bacillus subtilis) is found and reaches 99%. Thus, the strain was determined to be Bacillus subtilis. The strain obtained by screening is named as Bacillus subtilis KY 07.
Through identification, the bacillus subtilis KY07 can simultaneously degrade ammoniacal nitrogen and hydrogen sulfide in odor generated in the kitchen garbage fermentation process.
In a second aspect of the present invention, there is provided a fermentation method of said bacillus subtilis KY07, comprising:
inoculating the bacillus subtilis KY07 to a fermentation culture medium for fermentation culture; the fermentation medium comprises 5g/L of beef extract, 10g/L of protein, 5g/L of sodium thiosulfate, 6g/L of ammonium chloride and 0.05g/L of manganese sulfate.
Further, the temperature of the fermentation culture is 37 ℃; the fermentation culture conditions are aerobic.
In a third aspect of the present invention, there is provided a microbial inoculum, which comprises bacillus subtilis KY07 and/or a culture solution and/or a bacterial suspension thereof and/or a fermentation product thereof.
Further, the microbial inoculum also comprises a carrier.
In a specific embodiment, the microbial inoculum comprises a fermentation product of bacillus subtilis KY07 and a carrier.
Further, the number of viable bacteria in the microbial inoculum is 1.1 multiplied by 108～2.3×108cfu/g。
Further, the raw material of the carrier is selected from one or more than two of wood chips, bagasse powder, wheat bran, sawdust, straw scraps, reed scraps and bamboo powder.
Further, the preparation method of the carrier comprises the following steps: the raw materials are evenly mixed and then heated to kill harmful pathogenic bacteria.
In a specific embodiment, the raw materials of the carrier are wood chips and bagasse powder; the preparation method of the carrier comprises the steps of uniformly mixing the wood chips and bagasse powder according to the volume ratio of 2-3: 2-3, and heating at 60-80 ℃ to kill harmful pathogenic bacteria.
Further, the volume ratio of the wood chips to the bagasse powder is preferably 3: 2; the temperature of the heat treatment to kill harmful pathogenic bacteria is preferably 60 ℃.
The fourth aspect of the invention provides an application of the bacillus subtilis KY07 or a culture solution or a bacterial suspension thereof or a fermentation product thereof or the microbial inoculum in degrading ammonia nitrogen and/or hydrogen sulfide.
In a fifth aspect of the invention, the application of the bacillus subtilis KY07 or the culture solution or the bacterial suspension thereof or the fermentation product thereof or the microbial inoculum in reducing and/or eliminating kitchen garbage odor is provided.
Further, the odor is generated by biodegradation of kitchen garbage.
According to the sixth aspect of the invention, a method for reducing and/or eliminating kitchen garbage odor is provided, which comprises treating kitchen garbage with bacillus subtilis KY07 or a culture solution or a bacterial suspension thereof or a fermentation product thereof or the microbial inoculum.
Further, the condition of the treatment is aerobic.
Further, the conditions of the treatment are suitable for aerobic fermentation of the bacillus subtilis KY 07.
Further, the method for reducing and/or eliminating kitchen garbage odor comprises the following steps: (1) culturing the bacillus subtilis KY 07; (2) adsorbing the product of step (1) to a carrier; (3) mixing the product of step (2) with the kitchen waste to provide conditions suitable for aerobic fermentation of the bacillus subtilis KY 07.
Compared with the prior art, the invention has the following advantages:
the bacillus subtilis KY07 is obtained by screening, and can efficiently degrade odor generated in the fermentation process of kitchen garbage and reduce odor pollution. In the treatment process, only the bacillus subtilis KY07 is added, and no additional filler or energy is required to be added, so that the odor can be effectively reduced and/or eliminated. The bacillus subtilis KY07 provided by the invention obviously reduces the energy consumption of odor treatment, reduces the energy consumption cost in the process of kitchen garbage resource utilization, saves energy, reduces emission, and brings more economic values for the kitchen garbage resource utilization.
Detailed Description
Exemplary embodiments of the present disclosure will be described in more detail below. It should be understood that the present disclosure may be embodied in various forms and should not be limited by the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
Example 1
Obtaining a cattle manure sample in a cattle manure pile from a Jinhua Jiufeng pasture, and then inoculating the sample into a screening culture medium, wherein the screening culture medium is prepared as follows: 5.0g of beef extract, 10.0g of egg white, 5.0g of sodium thiosulfate, 5.0g of ammonium chloride and 1000ml of water, wherein the pH value is 7.0; 100ml of screening culture medium is prepared, a proper amount of sterile glass beads are added, 1.0g of cow dung sample is inoculated, and then enrichment is carried out for 7 days.
Inoculating the culture solution obtained by enrichment into a separation culture medium by a gradient dilution method, wherein the preparation of the separation culture medium is as follows: 10.0g of glucose, 6.0g of ammonium chloride, 5.0g of sodium thiosulfate, 4.0g of dipotassium hydrogen phosphate, 2.0g of monopotassium phosphate, 0.2g of magnesium chloride and 2ml of trace elements (4.0 g/L of zinc chloride, 5.0g/L of calcium chloride, 5.0g/L of manganese chloride, 5.0g/L of ferrous chloride, 1.0g/L of ammonium molybdate, 2.0g/L of copper chloride and 1.0g/L of cobalt chloride, wherein the pH is adjusted to 7.0), 1000ml of water and 2% of agar powder; and then selecting a single bacterial colony from the separation plate for streaking purification, repeatedly purifying for 2-3 times, performing streaking culture on the obtained pure bacterial colony, and keeping the pure bacterial colony in a refrigerator for later use, wherein the purified bacterial colony is bacillus subtilis KY 07.
Example 2
The preliminary test of ammonia nitrogen and hydrogen sulfide degradation of the strain obtained in example 1 is as follows:
preparing a liquid culture medium: 10.0g of glucose, 6.0g of ammonium chloride, 5.0g of sodium thiosulfate, 4.0g of dipotassium hydrogen phosphate, 2.0g of monopotassium phosphate, 0.2g of magnesium chloride and 2ml of trace elements (4.0 g/L of zinc chloride, 5.0g/L of calcium chloride, 5.0g/L of manganese chloride, 5.0g/L of ferrous chloride, 1.0g/L of ammonium molybdate, 2.0g/L of copper chloride and 1.0g/L of cobalt chloride, wherein the pH is adjusted to 7.0), and 1000ml of water. Adding 100ml of liquid culture medium into a 500ml triangular flask, and sterilizing for 20 minutes at 121 ℃ in an autoclave; 3 parallel experimental groups and 3 control groups were set, wherein 3 experimental groups were inoculated with the strain obtained in example 1 (the inoculum size was 1% of the culture medium), and 3 control groups were not inoculated with any strain.
After inoculation, the cells were incubated at 37 ℃ on a shaker at 200 rpm. The average data of 3 experimental groups were counted: under the condition, the ammonia nitrogen is reduced from 2.0g/L to 0.87g/L within 5 days, the degradation rate is 56.5 percent, and the concentration of sulfate ions generated after the sodium thiosulfate is degraded for 5 days is 1.1 g/L.
Therefore, the strain provided by the invention can degrade ammonia nitrogen and hydrogen sulfide simultaneously, has better degradation capability on ammonia gas and hydrogen sulfide in odor generated in the kitchen garbage fermentation process, and can effectively solve the odor problem in the kitchen garbage fermentation process.
Example 3
This example provides a fermentation process of Bacillus subtilis KY 07.
After the bacillus subtilis KY07 is subjected to strain activation, the bacillus subtilis KY07 is inoculated into a fermentation medium (5.0 g of beef extract, 10.0g of egg white, 5.0g of sodium thiosulfate, 6.0g of ammonium chloride, 0.05g of manganese sulfate and 1000ml of water, and the pH value is 7.0) and cultured at 37 ℃ and 200rpm for 24 hours to prepare the bacillus subtilis KY07 fermentation liquid.
Example 4
The embodiment provides a preparation method of a bacillus subtilis KY07 microbial inoculum.
Uniformly mixing the wood chips and bagasse powder in a volume ratio of 3 to 2, heating at 60 ℃ for 1h to kill harmful pathogenic bacteria, thus obtaining the carrier, and using the carrier in the preparation of bacillus subtilis KY07 microbial inoculum.
Spray drying and uniformly mixing the bacillus subtilis KY07 fermentation liquor prepared in the embodiment 3 with a carrier, and carrying out heat preservation treatment at 40 ℃ to obtain the bacillus subtilis KY07 microbial inoculum, wherein the viable count of the microbial inoculum is 1.1-2.3 multiplied by 108cfu/g。
Example 5
KY-WY100A biomass waste treatment equipment produced by two Kangyang environmental science and technology companies is adopted, one equipment uses a traditional microbial inoculum (namely the microbial inoculum provided by matching the biomass waste treatment equipment does not contain deodorant bacteria), and the other equipment adds the Bacillus subtilis KY07 microbial inoculum prepared in the embodiment 4 on the basis of the equivalent traditional microbial inoculum. Kitchen waste with the same source is added into the two devices respectively (the kitchen waste is added according to the mass ratio of 1:1 of the traditional microbial inoculum), the garbage treatment device is operated, and after 24 hours of garbage treatment, the concentration of ammonia gas and hydrogen sulfide is detected when the exhaust port of the deodorization device is exhausted.
Under the condition that the deodorization system is not started, the concentration of ammonia gas discharged from the exhaust port of the equipment using the traditional microbial inoculum is 4.7mg/m3The concentration of hydrogen sulfide is 0.34mg/m3. The concentration of ammonia gas discharged from the exhaust port of the equipment containing the bacillus subtilis KY07 is 1.8mg/m3The concentration of hydrogen sulfide was 0.17mg/m3. The bacillus subtilis KY07 can effectively reduce the emission of ammonia gas and hydrogen sulfide in the aerobic fermentation process of kitchen garbage.
After the deodorization system was turned on, the concentration of ammonia gas discharged from the exhaust port of the apparatus using the conventional microbial inoculum was 0.38mg/m3The concentration of hydrogen sulfide was 0.17mg/m3The energy consumption was 58 kwh/day. The concentration of ammonia gas discharged from the exhaust port of the equipment containing the bacillus subtilis KY07 is 0.31mg/m3Hydrogen sulfide concentration of 0.13mg/m3The energy consumption was 19 kwh/day. The bacillus subtilis KY07 can effectively reduce deodorization energy consumption of equipment and realize energy conservation and emission reduction in the operation process of the kitchen waste aerobic fermentation equipment.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (13)
1. Bacillus subtilis KY07, which is preserved in China center for type culture Collection and has a preservation number of CCTCC NO: M2020152.
2. The fermentation process of bacillus subtilis KY07 of claim 1, comprising:
inoculating the bacillus subtilis KY07 to a fermentation culture medium for fermentation culture; the fermentation medium comprises the following components: 5g/L beef extract, 10g/L egg white, 5g/L sodium thiosulfate, 6g/L ammonium chloride, 0.05g/L manganese sulfate and the balance of water.
3. The fermentation process of claim 2, wherein the temperature of the fermentation culture is 37 ℃.
4. The fermentation process of claim 2, wherein the fermentation culture conditions are aerobic.
5. A microbial inoculum comprising bacillus subtilis KY07 of claim 1;
the microbial inoculum also comprises a carrier; the carrier is prepared by uniformly mixing sawdust and bagasse powder in a volume ratio of 2-3: 2-3.
6. The microbial inoculum of claim 5, in which the viable count of the microbial inoculum is 1.1 x 108~2.3×108cfu/g。
7. The microbial inoculum according to claim 5 or 6, wherein the preparation method of the carrier comprises the following steps: the raw materials are evenly mixed and then heated to kill harmful pathogenic bacteria.
8. The microbial inoculum according to claim 5 or 6, wherein the carrier is prepared by uniformly mixing wood chips and bagasse powder in a volume ratio of 2-3: 2-3, and then heating at 60-80 ℃ to kill harmful pathogenic bacteria.
9. The application of the bacillus subtilis KY07 or the microbial inoculum of any one of claims 5 to 8 in degrading ammonia nitrogen and hydrogen sulfide.
10. A method for reducing ammonia nitrogen and hydrogen sulfide in kitchen waste odor, which is characterized by comprising the step of treating the kitchen waste with the bacillus subtilis KY07 according to claim 1 or the microbial inoculum according to any one of claims 5 to 8.
11. The method of reducing ammonia nitrogen and hydrogen sulfide in kitchen waste odor according to claim 10, wherein said treating conditions are aerobic.
12. The method for reducing ammonia nitrogen and hydrogen sulfide in kitchen waste odor according to claim 10, wherein said treating conditions are suitable for aerobic fermentation of said bacillus subtilis KY 07.
13. The method of reducing ammonia nitrogen and hydrogen sulfide in kitchen waste odors of claim 10, comprising: (1) culturing the bacillus subtilis KY 07; (2) adsorbing the product of step (1) to a carrier; (3) mixing the product of step (2) with the kitchen waste to provide conditions suitable for aerobic fermentation of the bacillus subtilis KY 07.
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| CN112831438B (en) * | 2021-01-25 | 2023-05-02 | 瑞燃(上海)环境工程技术有限公司 | Bacillus subtilis for deodorizing kitchen waste and application thereof |
| CN114107115A (en) * | 2021-11-30 | 2022-03-01 | 电子科技大学中山学院 | Composite microbial inoculum for efficiently degrading kitchen waste and preparation method thereof |
| CN113999803A (en) * | 2021-11-30 | 2022-02-01 | 电子科技大学中山学院 | Deodorizing microbial agent and preparation method thereof |
| CN114574383B (en) * | 2022-01-18 | 2024-02-23 | 金华康扬环境科技有限公司 | Efficient kitchen waste degradation composite microbial agent and preparation method and application thereof |
| CN114921356B (en) * | 2022-01-18 | 2024-04-02 | 金华康扬环境科技有限公司 | Household kitchen waste aerobic composting composite microbial agent and preparation method thereof |
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| CN102219309A (en) * | 2011-03-13 | 2011-10-19 | 沧州市中信生物科技有限公司 | SWater purifying agent with quick result |
| KR20170101431A (en) * | 2016-02-29 | 2017-09-06 | 주식회사 고려비엔피 | Probiotic composition for improving environment of livestock barn |
| CN110438024A (en) * | 2019-04-17 | 2019-11-12 | 浙江亲水园生物科技有限公司 | A kind of bacillus subtilis strain and its application |
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| CN102219309A (en) * | 2011-03-13 | 2011-10-19 | 沧州市中信生物科技有限公司 | SWater purifying agent with quick result |
| KR20170101431A (en) * | 2016-02-29 | 2017-09-06 | 주식회사 고려비엔피 | Probiotic composition for improving environment of livestock barn |
| CN110438024A (en) * | 2019-04-17 | 2019-11-12 | 浙江亲水园生物科技有限公司 | A kind of bacillus subtilis strain and its application |
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