A microbial deodorizing agent for Chinese medicine extract attached liquid, preparation method and application

Abstract

The present invention relates to the technical field of microbial deodorization, and in particular to a microbial deodorizing agent for Chinese medicine extract attached liquid, a preparation method and an application thereof. The microbial deodorizing agent for Chinese medicine extract attached liquid comprises the following raw materials in parts by weight: 45-50 parts of a composite microbial bacterial liquid and 30-45 parts of a composite adsorbent. The deodorizing agent provided by the present invention performs deodorization from multiple angles, wherein the composite microbial bacterial liquid can inhibit the growth of odor-producing microorganisms through "ecological occupation", and degrade odor molecules through its own growth and reproduction metabolism. Even if there is a small amount of odor that has not been removed, it can be covered by the fragrance generated in the metabolism process of the microorganism itself. The composite adsorbent with a porous structure is used in combination, so that malodorous gases such as ammonia and hydrogen sulfide in the environment can be decomposed and adsorbed, and the reproduction of pathogenic bacteria in the environment can be effectively inhibited. Therefore, the deodorizing agent finally prepared has broad application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of microbial deodorization, and in particular to a microbial deodorizing agent for Chinese medicine extract attached liquid, a preparation method and application thereof. Background Art

[0002] Traditional Chinese medicine wastewater contains a large amount of organic matter, trace elements, plant fiber and other components, and has great potential for resource utilization. At the same time, microbial deodorization and degradation composite bacterial agents have the advantages of being environmentally friendly and low-cost. They are currently an important research direction for the treatment of malodorous environments. Microbial deodorization utilizes the efficient adsorption, absorption and degradation of special microorganisms that can transform or degrade malodorous substances to purify sulfur-containing, nitrogen-containing and other malodorous gases generated or emitted during life and production, and convert malodorous components such as hydrogen sulfide, mercaptans and ammonia into harmless and odorless substances, thereby achieving the goal of improving environmental air quality and protecting health.

[0003] In recent years, there have been some studies in the field of deodorization at home and abroad. The deodorizing preparations reported include paraformaldehyde, zeolite, organic acid, enzyme preparation, urease inhibitor, microorganism, Chinese herbal medicine, etc. At present, there are many kinds of microbial deodorants on the market, but they are mainly single or compound lactic acid bacteria, and the deodorizing effect is not good. It is urgent to develop new efficient and safe composite microbial deodorizing and disinfecting products to meet the market demand for green development of agriculture and animal husbandry and welfare breeding.

[0004] The invention patent with publication number CN114672433B discloses a composite microbial deodorant, a preparation method and an application thereof. The invention obtains a composite microbial deodorant by compounding Loderma longisporum, Lactobacillus zeae, Lactobacillus plantarum and Lactobacillus paracasei. However, the deodorization range of the biological deodorant is small, and the effective ingredients in the composite microbial deodorant are easily lost in some high temperature or extreme environments, resulting in a relatively single application field.

[0005] Therefore, according to the above-mentioned related technologies, it is urgent to develop a microbial deodorizing agent for extracting the auxiliary liquid of traditional Chinese medicine, a preparation method and an application thereof. Summary of the invention

[0006] In view of this, the purpose of the present invention is to propose a microbial deodorant for Chinese medicine extract attached liquid, a preparation method and an application, so as to solve the problem that the application of deodorant agents in the prior art is relatively limited.

[0007] Based on the above purpose, the present invention provides a microbial deodorizing agent for Chinese medicine extract attached liquid, a preparation method and an application.

[0008] A microbial deodorizing agent for Chinese medicine extract attached liquid, comprising the following raw materials in parts by weight: 45-50 parts of composite microbial liquid, 30-45 parts of composite adsorbent;

[0009] The composite microbial liquid is a composite liquid of plant lactobacillus and aroma-producing yeast;

[0010] The composite adsorbent is obtained by mixing polymer-coated Lactobacillus plantarum and modified mesoporous silica spheres in a mass ratio of 20-30:10-15;

[0011] The polymer-encapsulated Lactobacillus plantarum is sodium alginate-pectin-encapsulated Lactobacillus plantarum;

[0012] The modified mesoporous silicon spheres are copper-doped mesoporous silica.

[0013] Preferably, the preparation method of the composite microbial liquid is as follows:

[0014] Step S1. Inoculate the aroma yeast in PDB medium for culture, and inoculate the obtained aroma yeast seed liquid into the corresponding fermentation material for fermentation for 2-3 days to obtain bacterial liquid 1:

[0015] Step S2. Inoculate Lactobacillus plantarum in MRS medium for culture, and inoculate the obtained Lactobacillus plantarum seed liquid into the corresponding fermentation material for fermentation for 2-3 days to obtain bacterial liquid 2:

[0016] Step S3. Mixing bacterial solution 1 and bacterial solution 2 to obtain a composite microbial bacterial solution;

[0017] The total number of viable bacteria of the plant lactobacillus and the aroma-producing yeast is 1000-10000 / mL.

[0018] Preferably, the preparation method of the polymer-coated Lactobacillus plantarum is as follows:

[0019] Step A1. Lactobacillus plantarum was isolated from Lactobacillus plantarum powder, and the strain was preserved at -80°C using 40% glycerol. The Lactobacillus plantarum strain was inoculated into 100 mL of MRS medium at a 1% inoculum amount, and cultured at 37°C for 16 hours. The culture solution was centrifuged at 8000 pm and 4°C for 10 minutes, the supernatant was discarded, and the Lactobacillus plantarum cells were resuspended and washed twice with 0.85% sterile saline, and finally resuspended to 2×10 10 CFU / mL for future use;

[0020] Step A2. Prepare sodium alginate solution and pectin solution with sterile distilled water, stir at 20-25°C for 100-120 min, mix the sodium alginate solution and pectin solution, and then add 2.5-3 mL 2×10 10CFU / mL of bacterial liquid, and then spray CaCl2 solution with a syringe, solidify for 25-30 minutes, filter, and finally resuspend in sterile water and wash 3-5 times to obtain polymer-coated Lactobacillus plantarum.

[0021] Preferably, the concentration of the sodium alginate solution in step A2 is 2%-3.5%, and the concentration of the pectin solution is 0.5%-2.5%.

[0022] Preferably, the volume ratio of the sodium alginate solution to the pectin solution in step A2 is 5-6:7-8

[0023] Preferably, the volume of the syringe in step A2 is 50 mL, the inner diameter is 2 cm, and the concentration of the CaCl2 solution is 1%-1.5%.

[0024] Preferably, the preparation method of the modified mesoporous silica spheres is as follows:

[0025] Step B1. Dissolve hexadecyltrimethylammonium bromide in deionized water, stir at 580-600 r / min at 20-25°C until dissolved, add concentrated ammonia and ethyl acetate, stir for 5-8 min, and obtain a mixed solution A;

[0026] Step B2. Add metal salt solution and ethylenediaminetetraacetic acid to the mixed solution A, then add ethyl orthosilicate, add deionized water after 5-10 minutes, stir for 22-24 hours, and then centrifuge at a speed of 3800-4000 r / min for 4-6 minutes, and pour off the supernatant to obtain solid B;

[0027] Step B3. Add solid B to anhydrous ethanol and ultrasonicate for 4-6 minutes. Centrifuge at 3800-4000 r / min to obtain a precipitate. Dry the precipitate at 75-80°C for 8-10 hours, then heat it to 550°C at a heating rate of 1-2°C / min and keep it warm for 4-5 hours to obtain modified mesoporous silica spheres.

[0028] Preferably, the usage ratio of hexadecyltrimethylammonium bromide, deionized water, concentrated aqueous ammonia and ethyl acetate in step B1 is 140-150 mg: 100-120 mL: 4-6 mL: 1.3-1.5 mL;

[0029] In step B2, the dosage ratio of the mixed solution A, the metal salt solution, tetraethyl orthosilicate and deionized water is 250-260 mL: 55-60 mL: 0.5-0.75 mL: 51-56 mL, the molar ratio of the metal ions in the ethylenediaminetetraacetic acid and the metal salt solution is 1-1.1: 1, the metal salt solution is any one of Zn(NO3)2·9H2O, Cu(NO3)2·3H2O and CuCl2·2H2O, and the concentration of the metal salt solution is 0.6-0.65 mg / mL;

[0030] The usage ratio of solid B and anhydrous ethanol in step B3 is 5-10 g: 50-60 mL.

[0031] A method for preparing a deodorizing agent made of microorganisms extracted from traditional Chinese medicine comprises the following steps: uniformly mixing a composite microorganism liquid and a composite adsorbent to obtain the deodorizing agent.

[0032] The invention discloses an application of a deodorizing agent made from microorganisms attached to traditional Chinese medicine extraction, wherein the deodorizing agent is used for decomposing malodorous gases in the environment and inhibiting the reproduction of pathogenic bacteria in the environment.

[0033] Beneficial effects of the present invention:

[0034] The deodorizing agent provided by the present invention can deodorize from multiple angles. The composite microbial liquid therein can inhibit the growth of odor-producing microorganisms through "ecological occupation" and degrade odor molecules through its own growth and reproduction metabolism. Even if there is a small amount of odor that has not been removed, it can be masked by the fragrance produced during the metabolism of the microorganisms themselves. The composite adsorbent with a porous structure can be used in combination to decompose and adsorb malodorous gases such as ammonia and hydrogen sulfide in the environment, and effectively inhibit the reproduction of pathogenic bacteria in the environment. In addition, since the composite microbial liquid in the present invention can be derived from traditional Chinese medicine wastewater, it can not only solve the problem of traditional Chinese medicine wastewater treatment, but also develop an efficient and environmentally friendly deodorizing agent. Therefore, the deodorizing agent finally prepared has broad application prospects. DETAILED DESCRIPTION

[0035] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments.

[0036] The plant lactobacillus used in the present invention is disclosed in the invention patent with the authorization announcement number of "CN114015586B" and the name of "a deodorizing and degrading composite bacterial agent and its preparation method and application", the deposit number is CGMCCNO.23388, and the deposit date is September 14, 2021;

[0037] The aroma-producing yeast used in the present invention is Pichia pastoris, which is disclosed in an invention patent with authorization announcement number "CN114015586B" and name "A deodorizing and degrading composite bacterial agent, its preparation method and application", with a preservation number of CGMCCNO.23387 and a preservation date of September 14, 2021.

[0038] Embodiment 1: A method for preparing a microbial deodorant agent of a Chinese medicine extract, comprising the following steps:

[0039] Step S1. Inoculate the aroma yeast in PDB medium for culture, and inoculate the obtained aroma yeast seed liquid into the corresponding fermentation material for fermentation for 2 days to obtain bacterial liquid 1:

[0040] Step S2. Inoculate Lactobacillus plantarum in MRS medium for culture, and inoculate the obtained Lactobacillus plantarum seed liquid into the corresponding fermentation material for fermentation for 2 days to obtain bacterial liquid 2:

[0041] Step S3. Mixing bacterial solution 1 and bacterial solution 2 to obtain a composite microbial bacterial solution, wherein the total number of viable bacteria of Lactobacillus plantarum and aroma yeast is 1000 / mL;

[0042] Step S4. Lactobacillus plantarum was isolated from the Lactobacillus plantarum powder, and the strain was preserved at -80°C using 40% glycerol. The Lactobacillus plantarum strain was inoculated into 100 mL of MRS medium at a 1% inoculum amount, and cultured at 37°C for 16 hours. The culture solution was centrifuged at 8000 pm and 4°C for 10 minutes, the supernatant was discarded, and the Lactobacillus plantarum cells were resuspended and washed twice with 0.85% sterile saline, and finally resuspended to 2×10 10 CFU / mL for future use;

[0043] Step S5. Prepare sodium alginate solution and pectin solution with sterile distilled water, stir them at 20°C for 100 min, mix 50 mL of sodium alginate solution and 70 mL of pectin solution, and add 2.5 mL of 2×10 10 CFU / mL of bacterial solution, and then sprayed with a 1% CaCl2 solution using a syringe with a volume of 50mL and an inner diameter of 2cm, solidified for 25min, filtered, and finally resuspended with sterile water and washed 3 times to obtain polymer-coated Lactobacillus plantarum, wherein the concentration of sodium alginate solution was 2% and the concentration of pectin solution was 0.5%;

[0044] Step S6. Dissolve 140 mg of hexadecyltrimethylammonium bromide in 100 mL of deionized water, stir at 580 r / min at 20°C until dissolved, add 4 mL of concentrated ammonia water and 1.3 mL of ethyl acetate, stir for 5 min, and obtain a mixed solution A;

[0045] Step S7. Add 55 mL of 0.6 mg / mL Zn(NO3)2·9H2O solution and ethylenediaminetetraacetic acid to 250 mL of mixed solution A, then add 0.5 mL of ethyl orthosilicate, add 51 mL of deionized water after 5 min, stir for 22 h, centrifuge at 3800 r / min for 4 min, pour off the supernatant, and obtain solid B, wherein the molar ratio of ethylenediaminetetraacetic acid to the metal ions in the metal salt solution is 1:1;

[0046] Step S8. Add 5 g of solid B to 50 mL of anhydrous ethanol and perform ultrasonic treatment for 4 min. Centrifuge at 3800 r / min to obtain a precipitate. Dry the precipitate at 75° C. for 8 h. Then heat the precipitate to 550° C. at a heating rate of 1° C. / min and keep the temperature for 4 h to obtain modified mesoporous silica spheres.

[0047] Step S9. 20 g of polymer-coated Lactobacillus plantarum and 10 g of modified mesoporous silica spheres are mixed evenly to obtain a composite adsorbent, and 45 g of the composite microbial culture liquid and 30 g of the composite adsorbent are mixed evenly to obtain a deodorizing bacterial agent.

[0048] Embodiment 2: A method for preparing a microbial deodorant agent of a Chinese medicine extract, comprising the following steps:

[0049] Step S1. Inoculate the aroma yeast in PDB medium for culture, and inoculate the obtained aroma yeast seed liquid into the corresponding fermentation material for fermentation for 2 days to obtain bacterial liquid 1:

[0050] Step S2. Inoculate Lactobacillus plantarum in MRS medium for culture, and inoculate the obtained Lactobacillus plantarum seed liquid into the corresponding fermentation material for fermentation for 2 days to obtain bacterial liquid 2:

[0051] Step S3. Mixing bacterial solution 1 and bacterial solution 2 to obtain a composite microbial bacterial solution, wherein the total number of viable bacteria of Lactobacillus plantarum and aroma yeast is 4000 / mL;

[0052] Step S4. Lactobacillus plantarum was isolated from the Lactobacillus plantarum powder, and the strain was preserved at -80°C using 40% glycerol. The Lactobacillus plantarum strain was inoculated into 100 mL of MRS medium at a 1% inoculum amount, and cultured at 37°C for 16 hours. The culture solution was centrifuged at 8000 pm and 4°C for 10 minutes, the supernatant was discarded, and the Lactobacillus plantarum cells were resuspended and washed twice with 0.85% sterile saline, and finally resuspended to 2×10 10 CFU / mL for future use;

[0053] Step S5. Sodium alginate solution and pectin solution were prepared with sterile distilled water, stirred at 22°C for 108 min, 55 mL of sodium alginate solution and 74 mL of pectin solution were mixed, and 2.7 mL of 2×10 10 CFU / mL of bacterial solution, and then sprayed with a 1.2% CaCl2 solution using a syringe with a volume of 50mL and an inner diameter of 2cm, solidified for 27min, filtered, and finally resuspended with sterile water and washed 4 times to obtain polymer-coated Lactobacillus plantarum, wherein the concentration of sodium alginate solution was 2.5% and the concentration of pectin solution was 1%;

[0054] Step S6. Dissolve 144 mg of hexadecyltrimethylammonium bromide in 108 mL of deionized water, stir at 586 r / min at 22°C until dissolved, add 5 mL of concentrated ammonia water and 1.35 mL of ethyl acetate, stir for 6 min, and obtain a mixed solution A;

[0055] Step S7. Add 56 mL of 0.62 mg / mL Cu(NO3)2·3H2O solution and ethylenediaminetetraacetic acid to 254 mL of mixed solution A, then add 0.6 mL of ethyl orthosilicate, add 53 mL of deionized water after 8 min, stir for 23 h, centrifuge at 3880 r / min for 5 min, pour off the supernatant, and obtain solid B, the molar ratio of ethylenediaminetetraacetic acid to the metal ions in the metal salt solution is 1.08:1;

[0056] Step S8. 7 g of solid B was added to 54 mL of anhydrous ethanol and ultrasonicated for 5 min. The mixture was centrifuged at 3900 r / min to obtain a precipitate. The precipitate was dried at 78° C. for 9 h, and then heated to 550° C. at a heating rate of 1.5° C. / min and kept warm for 4.4 h to obtain modified mesoporous silica spheres.

[0057] Step S9. 24 g of polymer-coated Lactobacillus plantarum and 12 g of modified mesoporous silica spheres are mixed evenly to obtain a composite adsorbent, and 47 g of composite microbial culture liquid and 35 g of the composite adsorbent are mixed evenly to obtain a deodorizing bacterial agent.

[0058] Embodiment 3: A method for preparing a microbial deodorant agent of a Chinese medicine extract, comprising the following steps:

[0059] Step S1. Inoculate the aroma yeast in PDB medium for culture, and inoculate the obtained aroma yeast seed liquid into the corresponding fermentation material for fermentation for 3 days to obtain bacterial liquid 1:

[0060] Step S2. Inoculate Lactobacillus plantarum in MRS medium for culture, and inoculate the obtained Lactobacillus plantarum seed liquid into the corresponding fermentation material for fermentation for 3 days to obtain bacterial liquid 2:

[0061] Step S3. Mixing bacterial solution 1 and bacterial solution 2 to obtain a composite microbial bacterial solution, wherein the total number of viable bacteria of Lactobacillus plantarum and aroma yeast is 7000 / mL;

[0062] Step S4. Lactobacillus plantarum was isolated from the Lactobacillus plantarum powder, and the strain was preserved at -80°C using 40% glycerol. The Lactobacillus plantarum strain was inoculated into 100 mL of MRS medium at a 1% inoculum amount, and cultured at 37°C for 16 hours. The culture solution was centrifuged at 8000 pm and 4°C for 10 minutes, the supernatant was discarded, and the Lactobacillus plantarum cells were resuspended and washed twice with 0.85% sterile saline, and finally resuspended to 2×10 10 CFU / mL for future use;

[0063] Step S5. Prepare sodium alginate solution and pectin solution with sterile distilled water, stir at 20-25°C for 116 min, mix 58 mL of sodium alginate solution and 78 mL of pectin solution, and add 2.8 mL of 2×10 10 CFU / mL of bacterial solution, and then sprayed with a 1.4% CaCl2 solution using a syringe with a volume of 50mL and an inner diameter of 2cm, solidified for 28min, filtered, and finally resuspended with sterile water and washed 4 times to obtain polymer-coated Lactobacillus plantarum, wherein the concentration of sodium alginate solution was 3% and the concentration of pectin solution was 1.5%;

[0064] Step S6. Dissolve 148 mg of hexadecyltrimethylammonium bromide in 116 mL of deionized water, stir at 592 r / min at 24°C until dissolved, add 5 mL of concentrated ammonia water and 1.4 mL of ethyl acetate, stir for 7 min, and obtain a mixed solution A;

[0065] Step S7. Add 59 mL of 0.63 mg / mL CuCl2·2H2O solution and ethylenediaminetetraacetic acid to 258 mL of mixed solution A, then add 0.7 mL of tetraethyl orthosilicate, add 55 mL of deionized water after 9 min, stir for 23 h, centrifuge at 3950 r / min for 6 min, pour off the supernatant, and obtain solid B, wherein the molar ratio of ethylenediaminetetraacetic acid to the metal ion in the metal salt solution is 1.08:1;

[0066] Step S8. 9 g of solid B was added to 58 mL of anhydrous ethanol and ultrasonicated for 5 min, and centrifuged at 3950 r / min to obtain a precipitate, and the precipitate was dried at 78° C. for 9 h, and then heated to 550° C. at a heating rate of 2° C. / min and kept warm for 5 h to obtain modified mesoporous silica spheres;

[0067] Step S9. 28 g of polymer-coated Lactobacillus plantarum and 13 g of modified mesoporous silica spheres are mixed evenly to obtain a composite adsorbent, and 48 g of composite microbial culture liquid and 40 g of the composite adsorbent are mixed evenly to obtain a deodorizing bacterial agent.

[0068] Embodiment 4: A method for preparing a microbial deodorant agent of a Chinese medicine extract, comprising the following steps:

[0069] Step S1. The aroma yeasts are inoculated into PDB medium for culture, and the obtained aroma yeast seed liquids are inoculated into corresponding fermentation materials for fermentation for 3 days to obtain bacterial liquid 1:

[0070] Step S2. Inoculate Lactobacillus plantarum in MRS medium for culture, and inoculate the obtained Lactobacillus plantarum seed liquid into the corresponding fermentation material for fermentation for 3 days to obtain bacterial liquid 2:

[0071] Step S3. Mixing bacterial solution 1 and bacterial solution 2 to obtain a composite microbial bacterial solution, wherein the total number of viable bacteria of Lactobacillus plantarum and aroma yeast is 10,000 / mL;

[0072] Step S4. Lactobacillus plantarum was isolated from the Lactobacillus plantarum powder, and the strain was preserved at -80°C using 40% glycerol. The Lactobacillus plantarum strain was inoculated into 100 mL of MRS medium at a 1% inoculum amount, and cultured at 37°C for 16 hours. The culture solution was centrifuged at 8000 pm and 4°C for 10 minutes, the supernatant was discarded, and the Lactobacillus plantarum cells were resuspended and washed twice with 0.85% sterile saline, and finally resuspended to 2×10 10 CFU / mL for future use;

[0073] Step S5. Prepare sodium alginate solution and pectin solution with sterile distilled water, stir them at 25°C for 120 min, mix 60 mL of sodium alginate solution and 80 mL of pectin solution, and add 3 mL of 2×10 10 CFU / mL of bacterial solution, and then sprayed with a 1.5% CaCl2 solution using a syringe with a volume of 50mL and an inner diameter of 2cm, solidified for 30min, filtered, and finally resuspended with sterile water and washed 5 times to obtain polymer-coated Lactobacillus plantarum, wherein the concentration of sodium alginate solution was 3.5% and the concentration of pectin solution was 2.5%;

[0074] Step S6. Dissolve 150 mg of hexadecyltrimethylammonium bromide in 120 mL of deionized water, stir at 600 r / min at 25°C until dissolved, add 6 mL of concentrated ammonia water and 1.5 mL of ethyl acetate, stir for 8 min, and obtain a mixed solution A;

[0075] Step S7. Add 60 mL of 0.65 mg / mL Cu(NO3)2·3H2O solution and ethylenediaminetetraacetic acid to 260 mL of mixed solution A, then add 0.75 mL of ethyl orthosilicate, add 56 mL of deionized water after 10 min, stir for 24 h, centrifuge at 4000 r / min for 6 min, pour off the supernatant, and obtain solid B, the molar ratio of ethylenediaminetetraacetic acid to the metal ions in the metal salt solution is 1.1:1;

[0076] Step S8. 10 g of solid B was added to 60 mL of anhydrous ethanol and ultrasonicated for 6 min, and centrifuged at 4000 r / min to obtain a precipitate, and the precipitate was dried at 80° C. for 10 h, and then heated to 550° C. at a heating rate of 2° C. / min and kept warm for 5 h to obtain modified mesoporous silica spheres;

[0077] Step S9. 30 g of polymer-coated Lactobacillus plantarum and 15 g of modified mesoporous silica spheres are mixed evenly to obtain a composite adsorbent, and 50 g of the composite microbial culture liquid and 45 g of the composite adsorbent are mixed evenly to obtain a deodorizing bacterial agent.

[0078] Comparative Example 1:

[0079] Compared with Example 1, the present comparative example did not add a composite adsorbent during the preparation of the deodorant bacterial agent, and the remaining steps and parameters were the same, which will not be repeated in this comparative example, and a deodorant was finally obtained.

[0080] Comparative Example 2:

[0081] Compared with Example 1, the present comparative example does not add polymer-encapsulated Lactobacillus plantarum during the preparation of the deodorant bacterial agent, and the remaining steps and parameters are the same, which will not be repeated in this comparative example, and finally a deodorant is obtained.

[0082] Comparative Example 3:

[0083] Compared with Example 1, the modified mesoporous silica spheres were not added in the preparation process of the deodorant bacterial agent in this comparative example, and the remaining steps and parameters were the same, which will not be repeated in this comparative example, and the deodorant was finally obtained.

[0084] Comparative Example 4:

[0085] Compared with Example 1, this comparative example only adjusts the "20g polymer-coated Lactobacillus plantarum and 10g modified mesoporous silica spheres" in the composite adsorbent to "10g polymer-coated Lactobacillus plantarum and 20g modified mesoporous silica spheres", and the remaining steps and parameters are the same, which will not be repeated in this comparative example, and finally a deodorant is obtained.

[0086] Performance test: The deodorants prepared in Examples 1 to 4 and Comparative Examples 1 to 4 were tested, and the specific operation steps are as follows:

[0087] Referring to the "Water and Wastewater Monitoring and Analysis Methods (Fourth Edition)", multiple groups of 40kg manure fermentation tanks were set up, and 3mL of the deodorant prepared in Example 1-Example 4 and Comparative Example 1-Comparative Example 4 were added to each group of fermentation tanks. Samples were taken after 5d, 10d and 20d, and the volatile solid (VS) concentration, ammonia concentration, hydrogen sulfide concentration, total volatile organic compound (TVOC) concentration, and comprehensive odor concentration (OU) concentration were measured using a gas rapid detector. The initial manure fermentation tank was used as a blank group. The results are shown in Tables 1, 2 and 3:

[0088] Table 1 Summary of test results when samples were added to the fermentation tank for 5 days in Examples 1-4 and Comparative Examples 1-4

[0089]

[0090] Table 2 Summary of test results when samples were added to the fermentation tank for 10 days in Examples 1-4 and Comparative Examples 1-4

[0091]

[0092] Table 3 Summary of test results when samples were added to the fermentation tank for 20 days in Examples 1-4 and Comparative Examples 1-4

[0093]

[0094] Data Analysis:

[0095] It can be seen from Table 1, Table 2 and Table 3 that the deodorant prepared by the present invention has a strong removal effect on volatile solids (VS), ammonia, hydrogen sulfide, total volatile organic compounds and comprehensive odor, and the removal effect increases with time. This is because the deodorizing bacteria agent provided by the present invention can deodorize from multiple angles. The composite microbial bacterial liquid can inhibit the growth of odor-producing microorganisms through "ecological occupation" and degrade odor molecules through its own growth and reproduction metabolism. Even if there is a small amount of odor that is not removed, it can be covered by the fragrance produced in the metabolism process of the microorganism itself. The composite adsorbent with a porous structure can remove malodorous gases such as ammonia and hydrogen sulfide in the environment. The composite adsorbent can decompose and adsorb the odor body and effectively inhibit the reproduction of pathogens in the environment. A small amount of modified mesoporous silica balls in the composite adsorbent are doped with metal ions, which can increase the adsorption capacity of mesoporous silica in the composite adsorbent for odors. At the same time, the presence of metal ions can also promote the composite microbial culture to play its role. The polymer-encapsulated Lactobacillus plantarum in an amount greater than that of the modified mesoporous silica balls can maintain the efficient deodorizing activity of Lactobacillus plantarum for a long time. However, in Comparative Example 4, when the amount of modified mesoporous silica balls is increased to twice the amount of polymer-encapsulated Lactobacillus plantarum, the excessive use of metal ions leads to the inactivation of some microorganisms, which weakens the effect of the deodorant.

[0096] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present invention is limited to these examples. Under the concept of the present invention, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

[0097] The present invention is intended to cover all such substitutions, modifications and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A microbial agent, It is characterized in that The raw materials include the following parts by weight: 45-50 parts of composite microbial liquid and 30-45 parts of composite adsorbent; The composite microbial liquid is a composite liquid of plant lactobacillus and aroma-producing yeast; The plant lactobacillus is deposited in China General Microbiological Culture Collection Center, the deposit address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCCNO.23388; The aroma-producing yeast is Pichia pastoris, with the accession number being CGMCCNO.23387; The composite adsorbent is obtained by mixing polymer-coated Lactobacillus plantarum and modified mesoporous silica spheres in a mass ratio of 20-30:10-15; The polymer-encapsulated Lactobacillus plantarum is a Lactobacillus plantarum encapsulated by a mixture of sodium alginate and pectin; The modified mesoporous silicon spheres are mesoporous silicon dioxide doped with divalent copper ions.

2. A microbial agent according to claim 1, It is characterized in that The preparation method of the composite microbial liquid is as follows: Step S1. Inoculate the aroma yeast in PDB medium for culture, and inoculate the obtained aroma yeast seed liquid into the corresponding fermentation material for fermentation for 2-3 days to obtain bacterial liquid 1: Step S2. Inoculate Lactobacillus plantarum in MRS medium for culture, and inoculate the obtained Lactobacillus plantarum seed liquid into the corresponding fermentation material for fermentation for 2-3 days to obtain bacterial liquid 2: Step S3. Mixing bacterial solution 1 and bacterial solution 2 to obtain a composite microbial bacterial solution; The total number of viable bacteria of the plant lactobacillus and the aroma-producing yeast is 1000-10000 / mL.

3. A microbial agent according to claim 1, It is characterized in that The preparation method of the polymer-coated plant lactobacillus is as follows: Step A1. Lactobacillus plantarum was isolated from Lactobacillus plantarum powder, and the strain was preserved at -80°C using 40% glycerol. The Lactobacillus plantarum strain was inoculated into 100 mL of MRS medium at a 1% inoculum amount, and cultured at 37°C for 16 h. The culture solution was centrifuged at 8000 pm and 4°C for 10 min, and the supernatant was discarded. The Lactobacillus plantarum cells were resuspended and washed twice with 0.85% sterile saline, and finally resuspended to 2×10 10 CFU / mL for future use; Step A2. Prepare sodium alginate solution and pectin solution with sterile distilled water, stir at 20-25°C for 100-120 min, mix the sodium alginate solution and pectin solution, and then add 2.5-3 mL 2×10 10 CFU / mL of bacterial solution, and then inject CaCl 2 The solution was solidified for 25-30 minutes, filtered, and finally resuspended with sterile water and washed 3-5 times to obtain polymer-coated Lactobacillus plantarum.

4. A microbial agent according to claim 3, It is characterized in that The concentration of the sodium alginate solution in step A2 is 2%-3.5%, and the concentration of the pectin solution is 0.5%-2.5%.

5. A microbial agent according to claim 3, It is characterized in that The volume ratio of the sodium alginate solution to the pectin solution in step A2 is 5-6:7-8.

6. A microbial agent according to claim 3, It is characterized in that The volume of the syringe in step A2 is 50 mL, the inner diameter is 2 cm, and the CaCl 2 The concentration of the solution is 1%-1.5%.

7. A microbial agent according to claim 1, It is characterized in that The preparation method of the modified mesoporous silica spheres is as follows: Step B1. Dissolve hexadecyltrimethylammonium bromide in deionized water, stir at 580-600 r / min at 20-25°C until dissolved, add concentrated ammonia and ethyl acetate, stir for 5-8 min, and obtain a mixed solution A; Step B2. Add metal salt solution and ethylenediaminetetraacetic acid to the mixed solution A, then add ethyl orthosilicate, add deionized water after 5-10 minutes, stir for 22-24 hours, and then centrifuge at a speed of 3800-4000 r / min for 4-6 minutes, and pour off the supernatant to obtain solid B; Step B3. Add solid B to anhydrous ethanol and ultrasonicate for 4-6 minutes. Centrifuge at 3800-4000 r / min to obtain a precipitate. Dry the precipitate at 75-80°C for 8-10 hours, then heat it to 550°C at a heating rate of 1-2°C / min and keep it warm for 4-5 hours to obtain modified mesoporous silica spheres.

8. A microbial agent according to claim 7, It is characterized in that The usage ratio of hexadecyltrimethylammonium bromide, deionized water, concentrated ammonia water and ethyl acetate in step B1 is 140-150 mg: 100-120 mL: 4-6 mL: 1.3-1.5 mL; The ratio of the mixed solution A, the metal salt solution, ethyl orthosilicate and deionized water in step B2 is 250-260 mL: 55-60 mL: 0.5-0.75 mL: 51-56 mL, the molar ratio of the ethylenediaminetetraacetic acid and the metal ions in the metal salt solution is 1-1.1: 1, and the metal salt solution is Zn(NO 3 ) 2 9H 2 O, Cu(NO 3 ) 2 ·3H 2 O and CuCl 2 ·2H 2 Any one of O, wherein the concentration of the metal salt solution is 0.6-0.65 mg / mL; The usage ratio of solid B and anhydrous ethanol in step B3 is 5-10 g: 50-60 mL.

9. A microbial agent according to any one of claims 1 to 8, It is characterized in that The preparation method of the microbial agent is as follows: the composite microbial liquid and the composite adsorbent are evenly mixed to obtain the microbial agent.

10. A microbial agent according to any one of claims 1 to 8, It is characterized in that The microbial agent is used to decompose malodorous gases in the environment and inhibit the reproduction of pathogenic bacteria in the environment.

Images