Microbial agent for degrading sulfur-containing malodorous substances as well as preparation method and application of microbial agent

Abstract

The invention discloses a microbial agent capable of degrading sulfur-containing malodorous substances. The microbial agent is prepared by compounding at least two of isothermal layer bacillus GDUTAN16, pseudomonas aeruginosa, enterococcus faecalis and rhodopseudomonas palustris according to a ratio of 50: 0: 0: 0-0: 0: 0: 50. The invention further discloses a preparation method of the microbial agent freeze-dried powder and application of the microbial agent freeze-dried powder in degradation of sulfur-containing malodorous substances in environments such as water, soil and atmosphere. The freeze-dried powder is prepared by freeze-drying a microbial agent and a freeze-drying protective agent according to a mass ratio of 1: 1-1: 50. According to the method, sulfur-containing malodorous substances can be specifically and efficiently removed, the effect of removing odor in water, soil, atmosphere and other environments is achieved, the treatment time can be shortened, and the method has the advantages of being good in effect, easy to implement, low in cost and the like; in addition, the freeze-dried powder is quick in microbial activity recovery after rehydration, easy to store and transport, high in activity after recovery and good in application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial degradation, and more specifically relates to a microbial agent for degrading sulfur-containing malodorous substances, a preparation method and application thereof. Background Art

[0002] Among urban domestic waste, kitchen waste accounts for the largest proportion (about 50%). Due to its high content of water, organic matter, oil and other characteristics, it is very easy to produce a large amount of smelly waste gas during its stacking, transportation, and landfill, such as hydrogen sulfide, ammonia, ethanol, acetaldehyde, propionaldehyde, ethyl acetate, methyl mercaptan, dimethyl disulfide, olefins, etc., which seriously affect people's daily life, threaten the living environment and physical and mental health of landfill workers and surrounding residents, and have attracted widespread public concern.

[0003] In the prior art, the methods for treating malodorous gases mainly include physical methods, chemical methods and biological methods. Among them, the biological method has been valued by environmental workers because of its advantages of low cost, good treatment effect and no secondary pollution. The biological method mainly uses the biochemical action of microorganisms to absorb and transform malodorous substances into nutrients for their own growth and metabolize them out of the body to achieve the purpose of removing odor. Microbial degradation of waste gas generally goes through three steps: 1) pollutants in the waste gas diffuse from the gas phase into the liquid phase; 2) pollutants dissolved in the liquid further diffuse into the biofilm due to the concentration difference, and are adsorbed and absorbed by the microorganisms therein; 3) pollutants entering the microbial cells are decomposed and utilized by microorganisms. For the malodorous waste gas released during garbage disposal, there are currently three main biological deodorization technologies used: biofiltration, biotrickling and biowashing. Among them, for the malodorous waste gas generated by kitchen waste treatment, the biotrickling method is mainly used, relying on the biofilm on the filler to decompose the malodorous substances into simple and harmless inorganic substances such as CO2 and H2O to eliminate the odor-causing components. It can be seen that the biofilm is the key to determining the deodorization effect.

[0004] However, there are currently few types of microbial agents that can be used to degrade malodorous waste gas, and they are all broad-spectrum agents that cannot specifically degrade malodorous substances. As a result, the acclimatization cycle and biofilm formation time are long, and the deodorization efficiency is low. Therefore, it is very necessary to develop a microbial agent that can efficiently degrade malodorous substances, has a simple production process, and is low in cost, to provide a new treatment method for the treatment of malodorous pollutants and enrich the microbial agent resource library. Summary of the invention

[0005] In order to overcome the above-mentioned defects and shortcomings in the prior art, the present invention provides a microbial agent for degrading sulfur-containing malodorous substances. The invention has a simple production process and low cost, and can greatly improve the treatment efficiency of malodorous substances, thereby providing a new and efficient microbial agent and method for treating malodorous pollutants and enriching the microbial agent resource library.

[0006] The second object of the present invention is to provide a method for preparing the microbial agent for degrading malodorous sulfur-containing substances which can degrade malodorous waste gas and sulfur-containing wastewater.

[0007] The third object of the present invention is to provide a freeze-dried powder of a microbial agent for degrading sulfur-containing malodorous substances prepared according to the above preparation method.

[0008] The fourth object of the present invention is to provide the application of the microbial agent for degrading sulfur-containing malodorous substances.

[0009] The above-mentioned object of the present invention is achieved through the following technical scheme: a microbial agent for degrading sulfur-containing malodorous substances is prepared by compounding at least two of Bacillus stratosphericus GDUTAN16, Pseudomonas aeruginosa, Enterococcus faecalis, and Rhodopseudomonas palustris.

[0010] Preferably, the stratosphere Bacillus (Bacillus stratosphericus)

[0011] The volume ratio of GDUTAN16, Pseudomonas aeruginosa, Enterococcus faecalis, and Rhodopseudomonas palustris is 50:0:0:0-0:0:0:50.

[0012] The Bacillus stratosphericus GDUTAN16 is deposited in the China Center for Type Culture Collection with a deposit number of CCTCC NO: M 20211470.

[0013] The Pseudomonas aeruginosa is screened from landfill leachate. The screening steps of Pseudomonas aeruginosa include: firstly, the strain is domesticated with dimethyl trisulfide as a domestication substrate, and the concentration of the domestication substrate dimethyl trisulfide is 1 mg / L, 5 mg / L, 10 mg / L, 15 mg / L, 20 mg / L and 25 mg / L in sequence; then, domestication, the domestication includes the steps of: firstly, taking 10 mL of landfill leachate and adding it to an inorganic salt culture medium containing 1 mg / L dimethyl trisulfide, and after domestication at a temperature of 37°C for 5 days, the inoculation amount is transferred to the next dimethyl trisulfide concentration for domestication, and the concentration is gradually increased for domestication by analogy; after the domestication is completed, the bacterial solution with different domestication substrate concentrations is diluted by 10 -1 ~10 -7 times, and then take 0.2 mL of each and evenly spread it on the inorganic salt culture medium with dimethyl trisulfide as the carbon source, and select single bacteria with fast growth rate and neat edges. The degradation experiment was carried out after the single colony was enriched and cultured, and the colony with the best degradation effect was preserved and identified as a strain, thereby obtaining the Pseudomonas aeruginosa of the present invention; the components and proportions of the inorganic salt culture medium include the following: K2HPO4·3H2O 0.001-1.2g / L, KH2PO4 0.001-1.2g / L, NH4Cl 0.001-0.4g / L, MgSO4·7H2O 0.001-0.2g / L, FeSO4·7H2O0.001-0.01g / L, CaCl2·2H2O 0.001-0.2g / L, MnSO4·4H2O 0.001-0.2g / L, CuSO4·2H2O 0.001-0.01g / L, ZnSO4·7H2O 0.001-0.2g / L, CaCl2·6H2O 0.001-0.09g / L, Na2MoO4·2H2O 0.001-0.12g / L, H3BO30.001-0.006g / L.

[0014] The Enterococcus faecalis can be purchased from Shanghai Bofeng Biotechnology Co., Ltd. with an ATCC number of ATCC 2921229212.

[0015] The Rhodopseudomonas palustris can be purchased from Beina Chuanglian Biotechnology Co., Ltd. in the mall, and the strain number is BNCC336448.

[0016] The preparation method of the microbial agent for degrading sulfur-containing malodorous substances comprises compounding the above-mentioned different types of sulfur-containing malodorous substance degrading bacteria for purifying sulfur-containing malodorous substances in the environment, and mixing the compounded microbial strains with a lyophilization protective agent to prepare a microbial lyophilized powder.

[0017] Preferably, the preparation method of the microbial agent for degrading sulfur-containing malodorous substances includes the following steps: picking up separated and purified single colonies of Bacillus stratosphericus, Pseudomonas aeruginosa, Enterococcus faecalis or Rhodopseudomonas palustris and inoculating them into liquid culture medium respectively, culturing them in a shaker at 0-37°C and a rotation speed of 0-200rpm for 12-24h to obtain a degradation bacterial fermentation liquid; centrifuging the bacterial fermentation liquid, discarding the supernatant, and collecting the microbial sludge; adding a lyophilization protective agent and vortexing, mixing thoroughly and freeze-drying to obtain a microbial agent.

[0018] The liquid culture medium is one of nutrient broth culture medium, LB culture medium, ATYP culture medium, MAC culture medium, PCA culture medium or TSA culture medium.

[0019] The preparation method of the lyophilized protective agent comprises the following steps: dissolving 1-10 g of lyophilized protective substance in 1-100 ml of sterile water to obtain lyophilized protective agent liquid.

[0020] The freeze-drying protective substance is at least one of skimmed milk powder, glycerol, rhamnolipid and mannitol.

[0021] The sterile water is prepared by sterilizing deionized water at 120° C. for 30 minutes.

[0022] Preferably, the mass ratio of the microbial sludge and the lyophilization protective agent when mixed is 1:1-1:50.

[0023] A freeze-dried powder of a microbial agent for degrading sulfur-containing malodorous substances. The microbial agent for degrading sulfur-containing malodorous substances is pre-frozen at -20 to -80°C for 0 to 6 hours and then freeze-dried at 0 to -40°C for 10 to 72 hours to obtain the microbial dry powder.

[0024] Preferably, the effective viable bacteria count of the freeze-dried powder of the microbial agent for degrading sulfur-containing malodorous substances is 1.0×10 6 -1×10 10 CFU / g.

[0025] The microbial agent for degrading sulfur-containing malodorous substances is used for removing sulfur-containing malodorous substances in the environment.

[0026] The microbial agent for degrading sulfur-containing malodorous substances is a composite microorganism, and its application in environmental purification such as water, soil and atmosphere should also be within the protection scope of the present invention.

[0027] Optionally, the application refers to the use of active microorganisms to remove sulfur-containing malodorous substances in the environment.

[0028] The environment is water, air or soil polluted by sulfur-containing malodorous substances.

[0029] The sulfur-containing malodorous substance is at least one sulfur-containing malodorous substance including dimethyl disulfide, dimethyl trisulfide, methyl mercaptan and hydrogen sulfide.

[0030] The microbial agent for degrading sulfur-containing malodorous substances of the present invention can grow and reproduce with the malodorous gas generated in the process of treating urban sludge, livestock and poultry feces, sewage treatment, domestic garbage, etc. as the only carbon source and energy source, and has good performance in degrading malodorous waste gas. In addition, the preparation method of the microbial agent for degrading sulfur-containing malodorous substances of the present invention is simple, low in cost, high in activity, strong in pertinence, significant in deodorizing effect, high in efficiency, wide in applicability, short in biofilm formation time and strong in stability when used in biological equipment, and has incomparable advantages over the commercially available agents.

[0031] Compared with the prior art, the beneficial effects of the present application are as follows: it can specifically and efficiently remove sulfur-containing malodorous substances in water bodies, soil, atmosphere and other environments, achieve the effect of removing odors, and can also shorten the processing time, with good effects, simplicity and low cost; in addition, the freeze-dried powder of the composite microbial agent prepared with the help of a freeze-dried protective agent can quickly restore the activity of the microorganisms after rehydration, so that the microbial agent has the advantages of easy storage and transportation, and strong activity after recovery, and has a good application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is a diagram showing the degradation effect of Example 1;

[0033] Figure 2 This is a diagram showing the degradation effect of Example 2;

[0034] Figure 3 This is a photo of a dry powder sample of a microbial agent for degrading sulfur-containing odorous substances.

[0035] Specific implementation method

[0036] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0037] Example 1 Degradation effect of stratosphere bacillus on dimethyl disulfide

[0038] The isolated and purified Bacillus stratosphericus GDUTAN16 was streaked on a nutrient agar medium and cultured at 37°C for 24 h. A single colony was picked with an inoculation loop and inoculated into 50 ml of nutrient broth medium. After inoculation, the culture was placed in a shaker at 37°C and 200 rpm for 12 h. The enrichment culture was used to obtain the OD 600 The fermentation liquid of the degradation bacteria is 1.0.

[0039] The microbial content in the microbial fermentation broth is about 1×10 9 CFU / ml. The composition and dosage (g / L) of the nutrient broth medium are: 3.0 beef extract, 10.0 peptone, 5.0 NaCl; the composition and dosage (g / L) of the nutrient agar medium are: 10.0 peptone, 3.0 beef extract powder, 5.0 sodium chloride, 15.0 agar.

[0040] Take 50 ml of the fermentation liquid of the degradation bacteria and 50 ml of the inorganic salt solution in a glass headspace bottle to mix to form a microbial degradation system, add 150 mg / L of dimethyl disulfide to the system, place it in a shaker at a temperature of 37 ° C and a rotation speed of 200 rpm, and degrade it for 0.5, 1, and 2 days. After that, take 1 ml of the headspace gas to determine the dimethyl disulfide content. The degradation effect is judged according to the dimethyl disulfide concentration. The degradation effect is as follows: Figure 1 As shown, 46.5% of dimethyl disulfide was removed after 2 days of treatment.

[0041] Wherein, the components and proportions of the inorganic salt solution are as follows: K2HPO4·3H2O 1.2 g / L, KH2PO4 1.2 g / L, NH4Cl 0.4 g / L, MgSO4·7H2O 0.2 g / L, FeSO4·7H2O 0.01 g / L, CaCl2·2H2O 0.2 g / L, MnSO4·4H2O 0.2 g / L, CuSO4·2H2O 0.01 g / L, ZnSO4·7H2O 0.2 g / L, CaCl2·6H2O 0.09 g / L, Na2MoO4·2H2O 0.12 g / L, H3BO3 0.006 g / L.

[0042] Example 2 Effect of microbial agents for degrading sulfur-containing malodorous substances on the degradation of dimethyl disulfide

[0043] The preparation method of the microbial agent for degrading sulfur-containing malodorous substances comprises the following steps: Bacillus stratosphericus GDUTAN16 and Pseudomonas aeruginosa are streaked and inoculated on a nutrient agar medium after separation and purification, and cultured at 37°C for 24 hours. Then, a single colony is picked up with an inoculation loop and inoculated into 50 ml of a nutrient broth medium. After inoculation, the culture is placed in a shaker at a temperature of 37°C and a rotation speed of 200 rpm for 12 hours, and an OD value is obtained by enrichment culture. 600 The fermentation liquid of the degradation bacteria is 1.0; the fermentation liquid of the degradation bacteria of the stratosphere Bacillus subtilis and the fermentation liquid of the degradation bacteria of the Pseudomonas aeruginosa are mixed in equal volumes of 1:1 to prepare a microbial agent for degrading sulfur-containing malodorous substances.

[0044] The microbial content in the fermentation liquid of the microstratosphere Bacillus degradation strain and the fermentation liquid of Pseudomonas aeruginosa degradation strain is about 1×10 9 CFU / ml. The composition and dosage (g / L) of the nutrient broth medium are: 3.0 beef extract, 10.0 peptone, 5.0 NaCl; the composition and dosage (g / L) of the nutrient agar medium are: 10.0 peptone, 3.0 beef extract powder, 5.0 sodium chloride, 15.0 agar.

[0045] Take 50 ml of the microbial agent for degrading sulfur-containing odorous substances and 50 ml of the inorganic salt solution in a glass headspace bottle to mix to form a microbial degradation system. Add 150 mg / L of dimethyl disulfide to the system and place it in a shaker at a temperature of 37°C and a rotation speed of 200 rpm. After degradation for 0.5, 1, and 2 days, take 1 ml of the headspace gas to determine the dimethyl disulfide content. The degradation effect is judged according to the dimethyl disulfide concentration. The degradation effect is as follows: Figure 2 As shown, 99.1% of dimethyl disulfide was removed after 2 days of treatment, and there was no rebound in the subsequent time.

[0046] Wherein, the components and proportions of the inorganic salt solution are as follows: K2HPO4·3H2O 1.2 g / L, KH2PO4 1.2 g / L, NH4Cl 0.4 g / L, MgSO4·7H2O 0.2 g / L, FeSO4·7H2O 0.01 g / L, CaCl2·2H2O 0.2 g / L, MnSO4·4H2O 0.2 g / L, CuSO4·2H2O 0.01 g / L, ZnSO4·7H2O 0.2 g / L, CaCl2·6H2O 0.09 g / L, Na2MoO4·2H2O 0.12 g / L, H3BO3 0.006 g / L.

[0047] Example 3 Effect of microbial agents for degrading sulfur-containing malodorous substances on the degradation of dimethyl disulfide

[0048] The preparation method of the microbial agent for degrading sulfur-containing malodorous substances comprises the following steps: Bacillus stratosphericus GDUTAN16, Pseudomonas aeruginosa and Enterococcus faecalis are streaked and inoculated on a nutrient agar medium after separation and purification, and cultured at 37°C for 24 hours; then, a single colony is picked up with an inoculation loop and inoculated into 50 ml of a nutrient broth medium; after inoculation, the culture is placed in a shaker at a temperature of 37°C and a rotation speed of 200 rpm for 12 hours; and an OD value is obtained by enrichment culture. 600 The fermentation liquid of the degradation bacteria is 1.0; the fermentation liquid of the degradation bacteria of the stratosphere Bacillus, the fermentation liquid of the degradation bacteria of the Pseudomonas aeruginosa and the fermentation liquid of the degradation bacteria of the Enterococcus faecalis are mixed in a volume ratio of 1:1:1 to prepare a microbial agent for degrading sulfur-containing malodorous substances.

[0049] The microbial content in the fermentation liquid of the stratosphere Bacillus degrading bacteria, the fermentation liquid of Pseudomonas aeruginosa degrading bacteria and the fermentation liquid of Enterococcus faecalis degrading bacteria is about 1×10 9 CFU / ml. The composition and dosage (g / L) of the nutrient broth medium are: 3.0 beef extract, 10.0 peptone, 5.0 NaCl; the composition and dosage (g / L) of the nutrient agar medium are: 10.0 peptone, 3.0 beef extract powder, 5.0 sodium chloride, 15.0 agar.

[0050] Take 50 ml of microbial agent for degrading sulfur-containing malodorous substances and 50 ml of inorganic salt solution and mix them in a glass headspace bottle to form a microbial degradation system. Add 150 mg / L of dimethyl disulfide to the system, place it in a shaker at a temperature of 37°C and a rotation speed of 200 rpm. After degradation for 0.5, 1, and 2 days, take 1 ml of headspace gas to determine the dimethyl disulfide content. The degradation effect is judged according to the dimethyl disulfide concentration. After 1 day of treatment, 98.4% of the dimethyl disulfide is removed, and there is no rebound in the subsequent time.

[0051] The components and proportions of the inorganic salt solution are as follows: K2HPO4·3H2O 1.2g / L, KH2PO4 1.2g / L, NH4Cl 0.4g / L, MgSO4·7H2O 0.2g / L, FeSO4·7H2O 0.01g / L, CaCl2·2H2O 0.2g / L, MnSO4·4H2O 0.2g / L, CuSO4·2H2O 0.01g / L, ZnSO4·7H2O 0.2g / L, CaCl2·6H2O 0.09g / L, Na2MoO4·2H2O 0.12g / L, H3BO3 0.006g / L.

[0052] Example 4 Degradation effect of microbial agents for degrading sulfur-containing malodorous substances on dimethyl trisulfide

[0053] The preparation steps of the microbial agent for degrading sulfur-containing malodorous substances are as follows: Bacillus stratosphericus GDUTAN16 and Pseudomonas aeruginosa after separation and purification are streaked on nutrient agar medium, cultured at 37°C for 24 hours, and then a single colony is picked up with an inoculation loop and inoculated into 50 ml nutrient broth medium. After inoculation, it is placed in a shaker at a temperature of 37°C and a rotation speed of 200 rpm for 12 hours, and the OD value is obtained by enrichment culture. 600 The fermentation liquid of the degradation bacteria is 1.0; the fermentation liquid of the degradation bacteria of the stratosphere Bacillus and the fermentation liquid of the degradation bacteria of the Pseudomonas aeruginosa are mixed in a volume ratio of 2:3 to prepare a microbial agent for degrading sulfur-containing malodorous substances.

[0054] The microbial content in the fermentation liquid of the stratosphere Bacillus degradation strain and the fermentation liquid of Pseudomonas aeruginosa degradation strain is about 1×10 9 CFU / ml. The composition and dosage (g / L) of the nutrient broth medium are: 3.0 beef extract, 10.0 peptone, 5.0 NaCl; the composition and dosage (g / L) of the nutrient agar medium are: 10.0 peptone, 3.0 beef extract powder, 5.0 sodium chloride, 15.0 agar.

[0055] Take 50 ml of microbial agent for degrading sulfur-containing odorous substances and 50 ml of inorganic salt solution and mix them in a glass headspace bottle to form a microbial degradation system. Add 150 mg / L of dimethyl trisulfide to the system, place it in a shaker at a temperature of 37°C and a rotation speed of 200 rpm. After degradation for 0.5, 1, and 2 days, take 1 ml of headspace gas to determine the dimethyl trisulfide content. The degradation effect is judged according to the concentration of dimethyl trisulfide. After 0.5 days of treatment, 98.4% of dimethyl trisulfide is removed, and there is no rebound in the subsequent time.

[0056] The components and proportions of the inorganic salt solution are as follows: K2HPO4·3H2O 1.2g / L, KH2PO4 1.2g / L, NH4Cl 0.4g / L, MgSO4·7H2O 0.2g / L, FeSO4·7H2O 0.01g / L, CaCl2·2H2O 0.2g / L, MnSO4·4H2O 0.2g / L, CuSO4·2H2O 0.01g / L, ZnSO4·7H2O 0.2g / L, CaCl2·6H2O 0.09g / L, Na2MoO4·2H2O 0.12g / L, H3BO3 0.006g / L.

[0057] Example 5 Degradation effect of microbial agents for degrading sulfur-containing malodorous substances on dimethyl trisulfide

[0058] The preparation steps of the microbial agent for degrading sulfur-containing malodorous substances are as follows: Bacillus stratosphericus GDUTAN16, Pseudomonas aeruginosa and Enterococcus faecalis are streaked and inoculated on a nutrient agar medium after separation and purification, and cultured at 37°C for 24 hours. Then, a single colony is picked up with an inoculation loop and inoculated into 50 ml of a nutrient broth medium. After inoculation, the culture is placed in a shaker at a temperature of 37°C and a rotation speed of 200 rpm for 12 hours, and the OD value is obtained by enrichment culture. 600 The degradation bacteria fermentation liquid is 1.0; the degradation bacteria fermentation liquid of stratosphere Bacillus, the degradation bacteria fermentation liquid of Pseudomonas aeruginosa and the degradation bacteria fermentation liquid of Enterococcus faecalis are mixed in a volume ratio of 6:9:10 to prepare a microbial agent for degrading sulfur-containing malodorous substances.

[0059] The microbial content in the fermentation liquid of the stratosphere Bacillus degradation strain and the fermentation liquid of Pseudomonas aeruginosa degradation strain is about 1×10 9 CFU / ml. The composition and dosage (g / L) of the nutrient broth medium are: 3.0 beef extract, 10.0 peptone, 5.0 NaCl; the composition and dosage (g / L) of the nutrient agar medium are: 10.0 peptone, 3.0 beef extract powder, 5.0 sodium chloride, 15.0 agar.

[0060] Take 50 ml of microbial agent for degrading sulfur-containing malodorous substances and 50 ml of inorganic salt solution and mix them in a glass headspace bottle to form a microbial degradation system. Add 150 mg / L of dimethyl trisulfide to the system, place it in a shaker at a temperature of 37°C and a rotation speed of 200 rpm. After degradation for 0.5, 1, and 2 days, take 1 ml of headspace gas to determine the dimethyl trisulfide content. The degradation effect is judged according to the concentration of dimethyl trisulfide. After 0.5 days of treatment, 98.8% of the dimethyl trisulfide is removed, and there is no rebound in the subsequent time.

[0061] The components and proportions of the inorganic salt solution are as follows: K2HPO4·3H2O 1.2 g / L, KH2PO4 1.2 g / L, NH4Cl 0.4 g / L, MgSO4·7H2O 0.2 g / L, FeSO4·7H2O 0.01 g / L, CaCl2·2H2O 0.2 g / L, MnSO4·4H2O 0.2 g / L, CuSO4·2H2O 0.01 g / L, ZnSO4·7H2O 0.2 g / L, CaCl2·6H2O 0.09 g / L, Na2MoO4·2H2O 0.12 g / L, H3BO3 0.006 g / L.

[0062] Example 6 Preparation of freeze-dried powder of microbial agent for degrading sulfur-containing malodorous substances

[0063] The preparation method of the freeze-dried powder of microbial agent for degrading sulfur-containing malodorous substances is as follows:

[0064] The first step is to prepare a microbial fermentation liquid, wherein the microorganisms used include Bacillus stratosphericus GDUTAN16, Pseudomonas aeruginosa and Enterococcus faecalis; 25 ml of the Bacillus stratosphericus fermentation liquid and 25 ml of the Pseudomonas aeruginosa fermentation liquid are preliminarily mixed to prepare a mixed liquid A, and then 25 ml of the mixed liquid and 25 ml of the Enterococcus faecalis fermentation liquid are mixed to obtain a mixed liquid B; wherein the number of effective viable microorganisms in the microbial fermentation liquid is 1×10 9 CFU / ml;

[0065] In the second step, 25 ml of the microbial mixture B was centrifuged at 12000 rpm for 5 min, and the supernatant was discarded to obtain the microbial sludge;

[0066] In the third step, 2 g of microbial sludge was mixed with 10 ml of 10% skim milk powder freeze-drying protective agent liquid, and the mixture was fully mixed and pre-frozen at -80°C for 6 h and then freeze-dried at -40°C for 48 h to prepare a freeze-dried powder of a microbial agent for degrading sulfur-containing malodorous substances, such as Figure 3 shown.

[0067] After rehydration, gradient dilution and plating verification showed that the number of effective live bacteria in the freeze-dried powder of the microbial agent for degrading sulfur-containing malodorous substances obtained in this example was 1×10 10 CFU / g.

[0068] The 10% skim milk powder freeze-dried protective agent liquid is prepared by adding 10.0 g skim milk powder to ultrapure water to 1.0 L, and sterilized at 121° C. for 30 min before use.

[0069] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A microbial agent for degrading sulfur-containing malodorous substances, characterized in that: The compound is prepared from at least two of the following: Bacillus sp. GDUTAN16, Pseudomonas aeruginosa, Enterococcus faecalis, and Rhodopseudomonas palustris.

2. A microbial agent for degrading sulfur-containing malodorous substances according to claim 1, characterized in that: The volume ratio of the stratosphere bacillus GDUTAN16, Pseudomonas aeruginosa, Enterococcus faecalis, and Rhodopseudomonas palustris is 50:0:0:0-0:0:0:

50.

3. A microbial agent for degrading sulfur-containing malodorous substances according to claim 2, characterized in that: The stratosphere Bacillus GDUTAN16 was deposited in the China Center for Type Culture Collection with a collection number of CCTCC NO: M 20211470; the faecalis Enterococcus was purchased from Shanghai Bofeng Biotechnology Co., Ltd. with an ATCC number of ATCC 2921229212; the palustris Rhodopseudomonas was purchased from Shangcheng Beina Chuanglian Biotechnology Co., Ltd. with a strain number of BNCC336448.

4. A method for preparing a microbial agent for degrading sulfur-containing malodorous substances according to any one of claims 1 to 3, characterized in that: The steps include: inoculating single colonies of stratosphere bacillus, pseudomonas aeruginosa, enterococcus faecalis and red pseudomonas palustris into liquid culture medium respectively, culturing for 12-24 hours in a shaker at 0-37°C and a rotation speed of 0-200rpm to obtain degradation bacteria fermentation liquid; centrifuging the bacteria fermentation liquid, discarding the supernatant, collecting microbial sludge; adding a freeze-drying protective agent, fully mixing and freeze-drying.

5. The method for preparing a microbial agent for degrading sulfur-containing malodorous substances according to claim 4, characterized in that: The liquid culture medium is one of nutrient broth culture medium, LB culture medium, ATYP culture medium, MAC culture medium, PCA culture medium or TSA culture medium.

6. The method for preparing a microbial agent for degrading sulfur-containing malodorous substances according to claim 4, characterized in that: The preparation method of the lyophilized protective agent comprises the following steps: dissolving 1-10 g of lyophilized protective substance in 1-100 ml of sterile water to obtain lyophilized protective agent liquid.

7. The method for preparing a microbial agent for degrading sulfur-containing malodorous substances according to claim 6, characterized in that: The freeze-dried protective substance is at least one of skimmed milk powder, glycerol, rhamnolipid and mannitol; the sterile water is deionized water sterilized at 120° C. for 30 minutes.

8. The method for preparing a microbial agent for degrading sulfur-containing malodorous substances according to claim 7, characterized in that: The mass ratio of the microbial sludge and the freeze-drying protective agent when mixed is 1:1-1:

50.

9. A freeze-dried powder of a microbial agent for degrading sulfur-containing malodorous substances, characterized in that: The composite microbial degradation agent is prepared by the preparation method of a microbial agent for degrading sulfur-containing malodorous substances as described in claim 8, which is pre-frozen at -20 to -80°C for 0-6h and then freeze-dried at 0--40°C for 10-72h.

10. A microbial agent for degrading sulfur-containing malodorous substances is used to remove sulfur-containing malodorous substances in the environment.

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