CN111821847A - Composite biological deodorant and preparation method thereof - Google Patents
Composite biological deodorant and preparation method thereof Download PDFInfo
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
- B01D53/84—Biological processes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L9/00—Disinfection, sterilisation or deodorisation of air
- A61L9/01—Deodorant compositions
- A61L9/013—Deodorant compositions containing animal or plant extracts, or vegetable material
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L9/00—Disinfection, sterilisation or deodorisation of air
- A61L9/14—Disinfection, sterilisation or deodorisation of air using sprayed or atomised substances including air-liquid contact processes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2251/00—Reactants
- B01D2251/95—Specific microorganisms
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2257/00—Components to be removed
- B01D2257/30—Sulfur compounds
- B01D2257/304—Hydrogen sulfide
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2257/00—Components to be removed
- B01D2257/40—Nitrogen compounds
- B01D2257/406—Ammonia
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2257/00—Components to be removed
- B01D2257/70—Organic compounds not provided for in groups B01D2257/00 - B01D2257/602
- B01D2257/702—Hydrocarbons
- B01D2257/7022—Aliphatic hydrocarbons
- B01D2257/7025—Methane
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02C—CAPTURE, STORAGE, SEQUESTRATION OR DISPOSAL OF GREENHOUSE GASES [GHG]
- Y02C20/00—Capture or disposal of greenhouse gases
- Y02C20/20—Capture or disposal of greenhouse gases of methane
Abstract
The invention discloses a composite biological deodorant, and relates to the field of deodorants. The composite biological deodorant disclosed by the invention comprises a composite biological deodorant which comprises tea seed cake extracting solution, tea seed shell extracting solution, enterococcus faecalis, bacillus megaterium, bacillus cereus, bacillus methylotrophicus and rhodopseudomonas palustris. The composite biological deodorant provided by the invention is simple to prepare, has lower cost, reasonably utilizes waste resources, and has the characteristics of quick deodorization, long deodorization time, good deodorization effect and wide application range.
Description
Technical Field
The invention belongs to the technical field of deodorizers, and particularly relates to a preparation method of a composite biological deodorizer, a deodorizer and an application method thereof.
Background
The malodor is a gaseous substance which stimulates olfactory organs to cause people to be unpleasant and damages living environment, not only enables bacteria which propagate a large number of diseases to multiply and multiply, but also directly generates strong stimulation to a respiratory system, a nervous system, a circulatory system and an endocrine system through the olfactory system, and becomes important environmental pollution. Malodors are one of seven typical public hazards (air pollution, malodors, noise, water pollution, vibration, soil pollution and foundation subsidence), the substances are various, the influence range is large, and the control is gradually paid attention to people.
Malodorous gases can be divided into five types, namely sulfur-containing compounds, nitrogen-containing compounds, halogens and derivatives, hydrocarbons and oxygen-containing organic substances according to the chemical components of the gases. However, most of malodorous gases are detected to contain ammonia and hydrogen sulfide as main components. At present, there are three main types of methods for treating malodorous gases, namely physical methods, chemical methods and biological methods. The physical deodorization method and the chemical deodorization method adsorb or decompose odor through some active groups or inhibit the growth of odor-producing microorganisms, thereby achieving the purpose of deodorization, and the method has strong selectivity to odor components, slower deodorization speed, higher deodorization cost and easy generation of secondary pollution. As a new technology for controlling malodorous gas pollution, the biological deodorant has the advantages of good effect, wide application range, simple required equipment, easy management, convenient maintenance, no secondary pollution and the like, is generally concerned in the research and application of malodorous prevention at home and abroad, and the deodorization by using plant extract and microorganisms is an economic, efficient and environment-friendly mode which is widely applied.
The plant deodorant is a natural plant extract which is formed by extracting essential oil, juice or extract from trees, flowers and plants and other vegetation through micro emulsification or other processes, and the odor components are absorbed or reacted through main active components, so that the environment-friendly natural odor removing method is formed. However, the existing plant deodorant mainly utilizes the extract of plants such as trees, flowers and plants, and the like, so that the vegetation needs to be collected and a large amount of byproducts are produced after the extract is obtained, the ecological environment is damaged, and the environmental pressure is caused. Tea seed meal and tea seed shells are used as byproducts of tea seeds after the tea seed oil is squeezed, wherein the byproducts contain flavone, tea saponin, polysaccharide, tannin and other substances, and are main active ingredients for deodorization in a plant deodorant, and the active ingredients are not efficiently and reasonably utilized for a long time and are mostly discarded, so that huge resource waste is caused. Therefore, the method has great and profound significance for utilizing the waste of the tea seed cake and the tea seed shell and changing waste into valuable. In the prior report, the tea seed cake and the tea seed shell are rarely used as plant deodorant.
The microbial deodorization is to decompose and convert some odor substances into odorless substances or inhibit the generation of the odor substances by using microbes with specific functions so as to achieve the aim of removing odor. The composite deodorant is formed by purposefully compounding two or more single deodorizing components, so that the deodorizing mechanisms of the two or more single deodorizing components complement each other, and the two or more single deodorizing components play a role in different aspects to generate the optimal deodorizing effect. At present, various types of composite deodorizers discovered by domestic and overseas researches are available, but the existing composite deodorizers are troublesome to prepare, short in action time, high in cost and narrow in application range. Chinese patent CN104548175B discloses a composite biological deodorant, which is prepared by compounding plant extract with deodorizing microorganisms with different functions, and has the advantages of quick deodorization and strong deodorization duration, but the deodorant is prepared by using the plant extract, although the source is wide, vegetation in the natural environment is needed, the ecological environment is damaged, the byproducts are required to be additionally treated, the treatment cost is increased, and certain burden is caused to the ecological environment.
Disclosure of Invention
The invention aims to provide a composite biological deodorant which is simple to prepare, low in cost, reasonable in resource utilization, fast in deodorization, long in deodorization time, good in deodorization effect and wide in application range.
In order to solve the technical problems, the invention provides a composite biological deodorant which comprises tea seed meal extracting solution, tea seed shell extracting solution, enterococcus faecalis, bacillus megaterium, bacillus cereus, bacillus methylotrophicus and rhodopseudomonas palustris.
Further, the specific preparation method of the tea seed cake extracting solution comprises the following steps: crushing tea seed meal, sieving the crushed tea seed meal with a 60-mesh sieve, soaking the crushed tea seed meal for 2 hours at the temperature of 60 ℃ in 90% ethanol, wherein the ratio of the tea seed meal to ethanol solution is 1 kg: 12L, then performing ultrasonic extraction at 30 ℃ for 1h, and recovering an ethanol solution from the extract at 60 ℃ under reduced pressure to obtain the tea seed cake extract.
Further, the specific preparation method of the tea seed shell extracting solution comprises the following steps: cleaning tea seed shells, drying, crushing, sieving by a 60-mesh sieve, soaking for 2 hours at the temperature of 60 ℃ in 90% ethanol, wherein the ratio of the tea seed shells to ethanol solution is 1 kg: 12L, then performing ultrasonic extraction at 25 ℃ for 40min, and recovering ethanol solution from the extract at 60 ℃ under reduced pressure to obtain the tea seed shell extract.
Furthermore, the viable count of the enterococcus faecalis in the composite biological deodorant is 4.1 multiplied by 108-5.0×108cfu/mL; the viable count of the bacillus megaterium in the composite biological deodorant is 6.7 multiplied by 107-8.5×107cfu/mL; the viable count of the bacillus cereus in the composite biological deodorant is 3.9 multiplied by 107-5.8×107cfu/mL; the viable count of the bacillus methylotrophicus in the composite biological deodorant is 8.9 multiplied by 107-1.4×108cfu/mL; the viable count of the rhodopseudomonas palustris in the composite biological deodorant is 2.8 multiplied by 108-4.7×108cfu/mL。
A preparation method of the composite microbial deodorant comprises the following steps:
(1) mixing the tea seed meal extracting solution and the tea seed shell extracting solution, and then diluting by 100 times with sterile water to obtain a solution A for later use;
(2) solid slant culture: inoculating enterococcus faecalis into a solid MRS culture medium, inoculating bacillus megaterium into a solid LB culture medium, inoculating bacillus cereus and bacillus methylotrophicus into a solid nutrient agar culture medium, and culturing at constant temperature of 30 ℃ for 1-2 days to activate the strains;
inoculating the rhodopseudomonas palustris in a solid LB culture medium, adding paraffin into the liquid surface of the solid LB culture medium for liquid surface sealing, and activating a bacterial strain after short-distance irradiation by a 25W incandescent lamp and anaerobic culture for 8-10 h;
(3) seed liquid culture: inoculating the activated enterococcus faecalis in the step (2) into an MRS culture medium, inoculating the activated bacillus megaterium into an LB culture medium, inoculating the bacillus cereus and the bacillus methylotrophicus into a nutrient agar culture medium, inoculating the activated rhodopseudomonas palustris into the LB culture medium, culturing for 2-3 days in a shaking table with the temperature of 100 plus materials and the speed of 150r/min at 30-32 ℃, and stopping culturing when the density OD600 value of liquid culture bacteria reaches 0.5-1.0 to obtain a seed solution of the required microbial inoculum;
(4) preparing fermentation liquor: mixing the enterococcus faecalis seed liquid obtained in the step (3) and the rhodopseudomonas palustris with the inoculation amount of 15-20%, then inoculating the mixture into a fermentation culture medium, and carrying out sealed fermentation culture for 5-7 days at the temperature of 25-30 ℃ to obtain a solution B;
mixing the bacillus megaterium seed liquid, the bacillus cereus seed liquid and the bacillus methylotrophicus seed liquid obtained in the step (3) according to the inoculation amount of 5-15%, then inoculating the mixture into a fermentation culture medium, and performing sealed fermentation culture for 5-7 days at the temperature of 25-30 ℃ to obtain a solution C;
(5) mixing the solution B and the solution C obtained in the step (4) according to the volume ratio of 2:1 to obtain a mixed solution D;
(6) and (3) mixing the solution A and the solution D obtained in the step (1) and the step (5) according to the volume ratio of (1-1.5) to (0.8-1.0) to obtain the composite biological deodorant.
Furthermore, the volume ratio of the tea seed meal extracting solution to the tea seed shell extracting solution is (1-2) to 1.
Further, the formula of the fermentation medium in the step (4) comprises the following components in parts by weight: 2.0 parts of brown sugar, 1.2 parts of bean cake powder, 0.18 part of magnesium sulfate, 0.24 part of urea and the balance of deionized water.
Further, the preparation conditions of the fermentation medium are as follows: aerobic condition, aeration quantity is 1.0vvm, temperature is 30 ℃, and stirring speed is 150 r/min.
The application method of the composite biological deodorant comprises the steps of diluting the composite biological deodorant by 50-80 times with water, and spraying the composite biological deodorant into air with odor by spraying in Hebei with the amount of 0.1-0.3L/m2。
The invention achieves the following beneficial effects:
1. the tea seed meal extracting solution and the tea seed shell extracting solution contain flavonoid, tea saponin, polysaccharide, tannin, caffeine, theobromine and other substances, the tannin contains catechin and other components, and-OH on a B ring of the flavonoid and the catechin has stronger polarity and is easy to break, and the flavonoid and the-OH are combined with polar gas molecules such as ammonia gas, hydrogen sulfide, methyl mercaptan and the like to generate new organic or inorganic compounds, so that the ammonia gas, the hydrogen sulfide, the methyl mercaptan and other gases are removed. Caffeine and theobromine are alkaloids which react with acidic substances in odor to deodorize. The tannin and the flavonoid can block the normal expression of bacterial protein, influence the structural composition of cells and the catalytic activity of enzyme, and finally cause the loss of the normal physiological function of bacteria, thereby achieving the bacteriostatic effect. The virus has simple structure and is sensitive to tannin, so the tea seed cake extract and the tea seed shell extract also have antiviral effect.
2. The invention utilizes the combined microbial inoculum to convert odor substances such as ammonia gas, hydrogen sulfide, methane and the like into chemical nitrogen and other substances, can quickly remove and cover the odor substances, has the inhibiting effect on harmful bacteria by the microbial inoculum and metabolites thereof, prevents organic substances from being converted into the odor substances, and simultaneously can utilize the odor substances to reproduce and synthesize self protein or secondary metabolites under aerobic conditions, thereby realizing long-acting deodorization.
3. The preparation method of the tea seed meal extracting solution and the tea seed shell extracting solution is prepared by soaking the tea seed meal extracting solution and the tea seed shell extracting solution in ethanol and then extracting the tea seed meal extracting solution and the tea seed shell extracting solution by ultrasonic waves, and the preparation method is quick and simple in operation, short in time and high in extraction rate.
4. The tea seed cake extracting solution is an active substance extracted from tea seed cake, the tea seed shell extracting solution is an active substance extracted from tea seed shells, the tea seed cake and the tea seed shells are byproducts of tea seeds after the tea seeds squeeze tea seed oil, most of the tea seed cake and the tea seed shells are discarded or used as fertilizers, and huge resource waste and treatment cost are caused. The invention takes the extracting solution extracted by processing the tea seed dregs and the tea seed shells as the deodorant, reasonably utilizes the waste, reduces the cost, saves the waste treatment cost, protects the environment and is an environment-friendly deodorant.
5. According to the deodorant disclosed by the invention, the tea seed meal extracting solution and the tea seed shell extracting solution are compounded with the microbial bacteria with different functions, the active substances in the tea seed meal extracting solution and the tea seed shell extracting solution can be quickly deodorized, the nutrient substances in the tea seed meal extracting solution and the tea seed shell extracting solution can provide nutrition for microbial bacteria propagation, and the characteristic of long deodorization duration is realized.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The composite biological deodorant of the present invention will be described with reference to the following specific examples.
The Enterococcus faecalis strain of the invention is Enterococcus faecalis (Enterococcus faecalis) preserved by Beijing Beinan institute of Biotechnology, and has the resource number: BNCC 102668.
The Bacillus megaterium strain is Bacillus megaterium (Bacillus megaterium) deposited by Beijing Beinanna institute of Biotechnology, and has the following resource number: BNCC 190686.
The Bacillus cereus strain is Bacillus cereus (Bacillus cereus) deposited by Beijing Beinan union of Industrial science and Biotechnology, and has the following resource number: BNCC 191738.
The Bacillus methylotrophicus strain is Bacillus methylotrophicus (Bacillus methylotrophicus) preserved by Beijing Naina institute of Biotechnology, and has the resource number: BNCC 231950.
The rhodopseudomonas palustris of the invention is rhodopseudomonas palustris (Bacillus methylotrophicus) preserved by Beijing Nanagawa union biotechnology research institute, and has the resource number: BNCC 231950.
The solid MRS culture medium of the invention is: 10g of peptone, 10g of beef extract, 5g of yeast extract, 20g of glucose, 801.0mL of tween, 2g of dipotassium phosphate, 5g of sodium acetate, 2g of diammonium citrate, 0.58g of magnesium sulfate heptahydrate, 0.25g of manganese sulfate monohydrate, 18.0g of agar, 1000mL of distilled water and pH of 6.2-6.6. Agar is not added in the components of the MRS culture medium, and other components are the same as the solid MRS culture medium.
The solid LB culture medium of the invention is: 10g of tryptone, 5g of yeast powder, 10g of sodium chloride, 15g of agar, 1000mL of distilled water and 70-7.2 of PH. Agar was not added to the LB medium components, and the other components were the same as the solid LB medium.
The solid nutrient agar culture medium of the invention is: 3g of beef extract, 5g of sodium chloride, 10g of peptone, 20g of agar, 1000Ml of distilled water and pH 7.0-7.4. The nutrient agar medium has no agar added, and other components are the same as those of the solid nutrient agar medium.
Example 1 composite biological deodorant F1
1. Consists of the following components:
extracting solution of tea seed cake;
the specific preparation method of the tea seed cake extracting solution comprises the following steps: crushing tea seed meal, sieving with a 60-mesh sieve, soaking for 2 hours at the temperature of 60 ℃ in 90% ethanol, wherein the ratio of the tea seed meal to ethanol solution is 1 kg: 12L, then performing ultrasonic extraction at 30 ℃ for 1h, and recovering ethanol solution from the extract under reduced pressure at 60 ℃ to obtain tea seed cake extract.
Extracting tea seed shells;
the specific preparation method of the tea seed shell extracting solution comprises the following steps: cleaning tea seed shells, drying, crushing, sieving with a 60-mesh sieve, soaking at 60 ℃ in 90% ethanol for 2h, wherein the ratio of the tea seed shells to the ethanol solution is 1 kg: 12L, then performing ultrasonic extraction at 25 ℃ for 40min, and recovering ethanol solution from the extract at 60 ℃ under reduced pressure to obtain the tea seed shell extract.
The viable count of enterococcus faecalis in the composite biological deodorant is 4.1 × 108-5.0×108cfu/mL;
The viable count of the bacillus megaterium in the composite biological deodorant is 6.7 multiplied by 107-8.5×107cfu/mL;
Wax-like bacillusThe number of viable bacteria in the composite biological deodorant is 3.9 × 107-5.8×107cfu/mL;
The viable count of the methylotrophic bacillus in the composite biological deodorant is 8.9 multiplied by 107-1.4×108cfu/mL;
The viable count of the rhodopseudomonas palustris in the composite biological deodorant is 2.8 multiplied by 108-4.7×108cfu/mL。
2. The preparation method of the compound microbial deodorant comprises the following steps:
(1) mixing the above tea seed cake extractive solution and tea seed shell extractive solution at volume ratio of 1:1, and diluting with sterile water 100 times to obtain solution A1.
(2) Solid slant culture: inoculating enterococcus faecalis into solid MRS culture medium, inoculating Bacillus megaterium into solid LB culture medium, inoculating Bacillus cereus and Bacillus methylotrophicus into solid nutrient agar culture medium, and culturing at 30 deg.C for 1-2 days to activate the strain.
Inoculating Rhodopseudomonas palustris in a solid LB culture medium, adding paraffin into the liquid surface of the solid LB culture medium for liquid surface sealing, performing short-distance irradiation by a 25W incandescent lamp, performing anaerobic culture for 8-10h, and activating the strain.
(3) Seed liquid culture: inoculating the activated enterococcus faecalis in the step (2) into an MRS culture medium, inoculating the activated bacillus megaterium into an LB culture medium, inoculating the bacillus cereus and the bacillus methylotrophicus into a nutrient agar culture medium, inoculating the activated rhodopseudomonas palustris into the LB culture medium, culturing for 2-3 days in a shaking table with the temperature of 100 plus materials and the speed of 150r/min at 30-32 ℃, and stopping culturing when the density OD600 value of liquid culture bacteria reaches 0.5-1.0 to obtain a seed solution of the required microbial inoculum.
(4) Preparing fermentation liquor: and (3) mixing the enterococcus faecalis seed solution obtained in the step (3) and the rhodopseudomonas palustris with the inoculation amount of 15-20%, then inoculating the mixture into a fermentation culture medium, and carrying out sealed fermentation culture for 5-7 days at the temperature of 25-30 ℃ to obtain a solution B1.
And (3) mixing the bacillus megaterium seed liquid, the bacillus cereus seed liquid and the bacillus methylotrophicus seed liquid obtained in the step (3) according to the inoculation amount of 5-15%, then inoculating the mixture into a fermentation culture medium, and carrying out sealed fermentation culture for 5-7 days at the temperature of 25-30 ℃ to obtain a solution C1.
(5) And (3) mixing the solution B1 and the solution C1 obtained in the step (4) in a volume ratio of 2:1 to obtain a mixed solution D1.
(6) And (3) mixing the solution A1 and the solution D1 obtained in the step (1) and the step (5) according to the volume ratio of 1:0.8 to obtain the compound biological deodorant F1.
The formula of the fermentation medium in the embodiment comprises the following components in parts by weight: 2.0 parts of brown sugar, 1.2 parts of bean cake powder, 0.18 part of magnesium sulfate, 0.24 part of urea and the balance of deionized water. The fermentation medium is prepared by mixing the components in parts by weight, and uniformly stirring under aerobic condition at the aeration rate of 1.0vvm and the temperature of 30 ℃ at the speed of 150 r/min.
Example 2 composite biological deodorant F2
1. Consists of the following components:
the composition of example 2 is the same as example 1, with specific reference to example 1.
2. The preparation method of the compound microbial deodorant comprises the following steps:
(1) mixing the above tea seed cake extractive solution and tea seed shell extractive solution at volume ratio of 2:1, and diluting with sterile water 100 times to obtain solution A2 for use.
(2) Solid slant culture: the preparation method in the step (2) is the same as that in the example 1, and the specific steps refer to the example 1.
(3) Seed liquid culture: the preparation method in the step (3) is the same as that in the example 1, and the specific steps refer to the example 1.
(4) Preparing fermentation liquor: the preparation method in the step (4) is the same as that in example 1, and referring to example 1 for the specific steps, solution B2 and solution C2 were obtained.
(5) And (3) mixing the solution B2 and the solution C2 obtained in the step (4) in a volume ratio of 2:1 to obtain a mixed solution D2.
(6) And (3) mixing the solution A2 and the solution D2 obtained in the steps (1) and (5) according to the volume ratio of 1:1 to obtain the compound biological deodorant F2.
The formulation and preparation method of the fermentation medium in this example 2 are the same as those in example 1, please refer to example 1.
Example 3 composite biological deodorant F3
1. Consists of the following components:
the composition of example 3 is the same as example 1, with specific reference to example 1.
2. The preparation method of the compound microbial deodorant comprises the following steps:
(1) mixing the above tea seed cake extractive solution and tea seed shell extractive solution at volume ratio of 1:1, and diluting with sterile water 100 times to obtain solution A3.
(2) Solid slant culture: the preparation method in the step (2) is the same as that in the example 1, and the specific steps refer to the example 1.
(3) Seed liquid culture: the preparation method in the step (3) is the same as that in the example 1, and the specific steps refer to the example 1.
(4) Preparing fermentation liquor: the preparation method in the step (4) is the same as that in example 1, and referring to example 1 for the specific steps, solution B3 and solution C3 were obtained.
(5) And (3) mixing the solution B3 and the solution C3 obtained in the step (4) in a volume ratio of 2:1 to obtain a mixed solution D3.
(6) And (3) mixing the solution A3 and the solution D3 obtained in the step (1) and the step (5) according to the volume ratio of 1.5:0.8 to obtain the compound biological deodorant F3.
The formulation and preparation method of the fermentation medium in this example 3 are the same as those in example 1, please refer to example 1.
Comparative example 1 composite biological deodorant F4
1. Consists of the following components:
comparative example 1 has the same composition as example 1, with specific reference to example 1.
2. The preparation method of the compound microbial deodorant comprises the following steps:
(1) solid slant culture: the preparation method in the step (2) is the same as that in the example 1, and the specific steps refer to the example 1.
(2) Seed liquid culture: the preparation method in the step (3) is the same as that in the example 1, and the specific steps refer to the example 1.
(3) Preparing fermentation liquor: the preparation method in the step (4) is the same as that in example 1, and referring to example 1 for the specific steps, solution B4 and solution C4 were obtained.
(4) And (3) mixing the solution B4 and the solution C4 obtained in the step (3) according to the volume ratio of 2:1 to obtain a mixed solution D4, namely obtaining the compound biological deodorant F4.
The formulation and preparation of the fermentation medium in comparative example 1 are the same as in example 1, please refer to example 1.
Application example 1 application of composite biological deodorant to fresh pig manure
5 sealed sheds with the same size are arranged in a certain pig farm, the length multiplied by the width multiplied by the height is 3m multiplied by 2m multiplied by 1m, fresh pig manure with the same mass is flatly laid at the same position in the sealed shed, 50ml of each of the compound biological deodorants F1, F2, F3 and F4 diluted by 50 times in the embodiment 1-3 is sprayed after being sealed for 30 minutes, 50ml of water is sprayed in a blank group, the ammonia gas and the hydrogen sulfide value are measured by an instrument, the human body feels the stink degree, and the test result is shown in table 1.
TABLE 1 Change in values of ammonia and hydrogen sulfide after treatment of fresh pig manure (unit: ppm)
Application example 2 application of composite biological deodorant to garbage transfer station
5 sealing sheds with the same size are arranged at a certain garbage transfer station, the length multiplied by the width multiplied by the height is 3m multiplied by 2m multiplied by 1m, garbage with the same mass is flatly laid at the same position in the sealing shed, 50ml of the compound biological deodorants F1, F2, F3 and F4 diluted by 80 times in the embodiment 1-3 are sprayed after sealing for 30 minutes, 50ml of water is sprayed in a blank group, the ammonia gas and the hydrogen sulfide value are measured by an instrument, the human body feels the odor degree, and the test result is shown in table 2.
TABLE 2 Change of ammonia and hydrogen sulfide values after garbage disposal in the transfer station (unit: ppm)
Example 3 application of composite biological deodorant to composting site
5 sealing sheds with the same size are arranged in a certain composting field, the length multiplied by the width multiplied by the height is 3m multiplied by 2m multiplied by 1m, the garbage with the same mass is flatly paved at the same position in the sealing shed, 50ml of the compound biological deodorants F1, F2, F3 and F4 diluted by 60 times in the examples 1-3 are sprayed after sealing for 30 minutes, 50ml of water is sprayed in the blank group, the ammonia gas and the hydrogen sulfide value are measured by an instrument, the human body feels the stink degree, and the test result is shown in the table 3.
TABLE 3 variation of values of ammonia and hydrogen sulfide after treatment in composting site (unit: ppm)
From the experimental results of the application examples 1-3, the composite biological deodorant disclosed by the invention has a good deodorization effect when being applied to fresh pig manure, garbage transfer stations and composting farms, and is wide in application range, fast in deodorization and long in deodorization duration. The invention has better deodorization effect and quick deodorization after the tea seed cake extracting solution and the tea seed shell extracting solution are added.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.
Claims (9)
1. The composite biological deodorant is characterized by comprising tea seed cake extracting solution, tea seed shell extracting solution, enterococcus faecalis, bacillus megaterium, bacillus cereus, bacillus methylotrophicus and rhodopseudomonas palustris.
2. The compound biological deodorant according to claim 1, wherein the tea seed cake extract is prepared by a specific method comprising: crushing tea seed meal, sieving the crushed tea seed meal with a 60-mesh sieve, soaking the crushed tea seed meal for 2 hours at the temperature of 60 ℃ in 90% ethanol, wherein the ratio of the tea seed meal to ethanol solution is 1 kg: 12L, then performing ultrasonic extraction at 30 ℃ for 1h, and recovering an ethanol solution from the extract at 60 ℃ under reduced pressure to obtain the tea seed cake extract.
3. The compound biological deodorant according to claim 1, wherein the tea seed shell extract is prepared by a specific method comprising: cleaning tea seed shells, drying, crushing, sieving by a 60-mesh sieve, soaking for 2 hours at the temperature of 60 ℃ in 90% ethanol, wherein the ratio of the tea seed shells to ethanol solution is 1 kg: 12L, then performing ultrasonic extraction at 25 ℃ for 40min, and recovering ethanol solution from the extract at 60 ℃ under reduced pressure to obtain the tea seed shell extract.
4. The composite biological deodorant according to claim 1, wherein the viable count of enterococcus faecalis in the composite biological deodorant is 4.1 x 108-5.0×108cfu/mL;
The viable count of the bacillus megaterium in the composite biological deodorant is 6.7 multiplied by 107-8.5×107cfu/mL;
The viable count of the bacillus cereus in the composite biological deodorant is 3.9 multiplied by 107-5.8×107cfu/mL;
The viable count of the bacillus methylotrophicus in the composite biological deodorant is 8.9 multiplied by 107-1.4×108cfu/mL;
The viable count of the rhodopseudomonas palustris in the composite biological deodorant is 2.8 multiplied by 108-4.7×108cfu/mL。
5. A method for preparing a composite microbial deodorant according to any one of claims 1 to 4, comprising the steps of:
(1) mixing the tea seed meal extracting solution and the tea seed shell extracting solution, and then diluting by 100 times with sterile water to obtain a solution A for later use;
(2) solid slant culture: inoculating enterococcus faecalis into a solid MRS culture medium, inoculating bacillus megaterium into a solid LB culture medium, inoculating bacillus cereus and bacillus methylotrophicus into a solid nutrient agar culture medium, and culturing at constant temperature of 30 ℃ for 1-2 days to activate the strains;
inoculating the rhodopseudomonas palustris in a solid LB culture medium, adding paraffin into the liquid surface of the solid LB culture medium for liquid surface sealing, and activating a bacterial strain after short-distance irradiation by a 25W incandescent lamp and anaerobic culture for 8-10 h;
(3) seed liquid culture: inoculating the activated enterococcus faecalis in the step (2) into an MRS culture medium, inoculating the activated bacillus megaterium into an LB culture medium, inoculating the bacillus cereus and the bacillus methylotrophicus into a nutrient agar culture medium, inoculating the activated rhodopseudomonas palustris into the LB culture medium, culturing for 2-3 days in a shaking table with the temperature of 100 plus materials and the speed of 150r/min at 30-32 ℃, and stopping culturing when the density OD600 value of liquid culture bacteria reaches 0.5-1.0 to obtain a seed solution of the required microbial inoculum;
(4) preparing fermentation liquor: mixing the enterococcus faecalis seed liquid obtained in the step (3) and the rhodopseudomonas palustris with the inoculation amount of 15-20%, then inoculating the mixture into a fermentation culture medium, and carrying out sealed fermentation culture for 5-7 days at the temperature of 25-30 ℃ to obtain a solution B;
mixing the bacillus megaterium seed liquid, the bacillus cereus seed liquid and the bacillus methylotrophicus seed liquid obtained in the step (3) according to the inoculation amount of 5-15%, then inoculating the mixture into a fermentation culture medium, and performing sealed fermentation culture for 5-7 days at the temperature of 25-30 ℃ to obtain a solution C;
(5) mixing the solution B and the solution C obtained in the step (4) according to the volume ratio of 2:1 to obtain a mixed solution D;
(6) and (3) mixing the solution A and the solution D obtained in the step (1) and the step (5) according to the volume ratio of (1-1.5) to (0.8-1.0) to obtain the composite biological deodorant.
6. The preparation method of the composite biological deodorant according to claim 5, wherein the volume ratio of the tea seed meal extracting solution to the tea seed shell extracting solution is (1-2): 1.
7. The preparation method of the compound biological deodorant according to claim 5, wherein the formula of the fermentation medium in the step (4) is as follows according to the weight portion: 2.0 parts of brown sugar, 1.2 parts of bean cake powder, 0.18 part of magnesium sulfate, 0.24 part of urea and the balance of deionized water.
8. The method for preparing a composite biological deodorant according to claim 7, wherein the preparation conditions of the fermentation medium are as follows: aerobic condition, aeration quantity is 1.0vvm, temperature is 30 ℃, and stirring speed is 150 r/min.
9. The method for applying the composite biological deodorant as claimed in any one of claims 1 to 3, wherein the composite biological deodorant is diluted 50 to 80 times with water and sprayed in the air with odor in Hebei with a spray amount of 0.1 to 0.3L/m2。
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