A long-lasting antibacterial deodorant for sewage and preparation method thereof

Abstract

The present invention relates to the field of sewage treatment technology, and more particularly to a long-lasting antibacterial deodorant for sewage and a preparation method thereof. The long-lasting antibacterial deodorant for sewage is prepared from raw materials comprising the following components in parts by weight: 0.2-0.3 parts of 1,2-benzisothiazolin-3-one, 0.3-0.5 parts of p-tert-amylphenol, 0.1-0.2 parts of methyl parahydroxybenzoate, 4-6 parts of activated carbon, 4-6 parts of modified sustained-release microspheres, 8-12 parts of a modified polymer, and 2-3 parts of modified lysozyme.

Description

Technical Field

[0001] The present invention relates to the technical field of sewage treatment, and in particular to a long-acting antibacterial deodorant for sewage and a preparation method thereof. Background Art

[0002] With the continuous advancement of technology, the popularity of cleaning appliances such as sweeping robots and floor scrubbers is rapidly increasing. Most of these appliances are equipped with fresh water tanks and waste water tanks. The waste water tank is primarily responsible for recovering wastewater from sweeping and mopping. This recovered wastewater often contains dirt, hair, oil, milk, leftovers, and other contaminants. These contaminants, along with the humid water environment, provide ample conditions for the rapid growth of bacteria and mold, which can easily cause decay and produce sour and unpleasant odors. The presence of unpleasant odors and a large number of pathogenic bacteria can affect mood and even physical health. Existing deodorization methods commonly include fragrance masking, porous material adsorption, and chemical solution reaction. Fragrance masking often uses chemical or natural fragrances to mask odors, while porous material adsorption often uses activated carbon, molecular sieves, porous ceramics, and other materials to absorb odors. However, these two methods cannot eradicate odors and harmful bacteria; they can only reduce the olfactory stimulation of odors to a certain extent. The chemical solution reaction method mostly uses oxidizing fungicides (such as 84 disinfectant, potassium permanganate) or quaternary ammonium fungicides (such as benzalkonium chloride, decanium chloride). This method can eradicate odors and harmful bacteria, but it is a one-time sterilization method and fungicides need to be added again every time the water is changed, and it cannot achieve long-term antibacterial and deodorizing effects. Summary of the Invention

[0003] In view of the shortcomings of the existing technology, the present invention provides a long-lasting antibacterial deodorant for sewage and a preparation method thereof.

[0004] The present invention is achieved through the following technical solutions:

[0005] A long-lasting antibacterial deodorant for sewage is prepared by using raw materials comprising the following components in parts by weight: 0.2-0.3 parts of 1,2-benzisothiazolin-3-one, 0.3-0.5 parts of p-tert-amylphenol, 0.1-0.2 parts of methyl parahydroxybenzoate, 4-6 parts of activated carbon, 4-6 parts of modified sustained-release microspheres, 8-12 parts of modified polymers, and 2-3 parts of modified lysozyme.

[0006] Furthermore, the raw materials for preparing the modified sustained-release microspheres include the following components in parts by weight: 3-4 parts of Artemisia annua, 2-3 parts of licorice, 1-1.5 parts of jasmine, 1-2 parts of ginkgo leaves, 2-3 parts of cloves, 1-1.5 parts of Echinacea, 2-3 parts of dodecylamine (DDA), 5-7.5 parts of calcium nitrate tetrahydrate (CNT), and 6-10 parts of dopamine hydrochloride (DA).

[0007] Furthermore, the preparation method of the modified sustained-release microspheres comprises the following steps:

[0008] L1. Artemisia annua, licorice, jasmine, ginkgo biloba, cloves, and echinacea were dried and ground through a 60-mesh sieve. The mixture was added to deionized water at a ratio of 1 g:10-15 mL. The mixture was sonicated at 80 Hz for 20-30 min, filtered, and the filtrate was concentrated under reduced pressure to 20% of its original volume and freeze-dried to obtain the plant extract.

[0009] L2. Mix deionized water and anhydrous ethanol, add DDA, and dissolve with stirring at 40°C. Then, add tetraethyl orthosilicate (TEOS), triethyl phosphate (TEP), and CNTs dissolved in deionized water, sequentially, every 30 minutes. Stir and react for 3 hours. Let the mixture stand for 18-22 hours, centrifuge at 10,000 rpm for 10-15 minutes, wash the precipitate with deionized water and ethanol, freeze-dry, calcinate at 650°C for 3 hours, cool naturally, and grind into a fine powder to obtain porous microspheres.

[0010] L3. DA was dissolved in 10 mM Tris-HCl buffer (pH 8.5) at a concentration of 10 mg / mL to obtain a DA solution. The plant extract and porous microspheres were added to deionized water. After ultrasonic dispersion, the mixture was stirred at room temperature for 24 h, centrifuged at 10,000 rpm for 10-15 min, and the precipitate was dried at 50°C. The solution was added to the DA solution, stirred at 150-200 rpm for 24 h, and centrifuged. The precipitate was washed with deionized water and ethanol and dried to obtain modified sustained-release microspheres.

[0011] Furthermore, in step L2, the volume ratio of deionized water to anhydrous ethanol is 5:16.

[0012] Furthermore, in step L2, the ratio of DDA to anhydrous ethanol is 1 g:20 mL.

[0013] Furthermore, in step L2, the volume ratio of TEOS, TEP and anhydrous ethanol is 10:1:50.

[0014] Furthermore, in step L2, the mass concentration of the CNT in deionized water is 0.5 g / mL.

[0015] Furthermore, the raw materials for preparing the modified polymer include the following components in parts by weight: 1-1.5 parts of 3-acrylamidophenylboronic acid (AAPBA), 4-6 parts of acrylamide (AM), 1.5-2.5 parts of N-(3-aminopropyl)acrylamide (NAPA), and 1.2-1.8 parts of 3-hydroxysuccinimide maleimidopropionate (SMMP).

[0016] Furthermore, the preparation method of the modified polymer comprises the following steps:

[0017] V1. Add AM, AAPBA, and NAPA to the DMSO solution and stir thoroughly. Vacuum-freeze-thaw-degassed the mixture twice. Add azobisisobutyronitrile (AIBN), seal the container, vacuum-freeze-thaw-degassed the mixture again, and react at 60°C for 10-12 hours. Cool to room temperature, add 10 volumes of ethanol, filter, wash the filter cake with ethanol, and freeze-dry.

[0018] V2. The product obtained in step V1 was added to PBS buffer (pH 7.4) and mixed. SMMP was added and stirred at room temperature at 200-300 rpm for 3-4 h. The product was dialyzed against deionized water (MWCO 5-7 kDa) for 24 h and freeze-dried to obtain a modified polymer.

[0019] Furthermore, in step V1, the mass concentration of AM in DMSO is 20 mg / mL.

[0020] Furthermore, in step V1, the amount of AIBN used is 0.5 wt%-0.6 wt% of AM.

[0021] Furthermore, in step V2, the mass concentration of the SMMP in the PBS buffer solution is 2 mg / mL.

[0022] Furthermore, the preparation method of the modified lysozyme comprises the following steps:

[0023] X1. Dissolve lysozyme in PBS containing 4 mM EDTA, pH 8.0, to obtain a lysozyme solution. Add 2-iminothiolane hydrochloride (Traut's reagent) to PBS containing 4 mM EDTA, pH 8.0, to obtain a Traut's reagent solution.

[0024] X2. Mix the Traut's reagent solution obtained in step X1 with the lysozyme solution. Aerate the reaction system with nitrogen and protect from light. Stir at 150 rpm for 1-2 h. Dialyze against PBS buffer at 4°C and freeze-dry to obtain the modified lysozyme.

[0025] Furthermore, in step X1, the mass concentration of the lysozyme in the PBS buffer solution is 2 mg / mL.

[0026] Furthermore, in step X1, the mass concentration of the Traut's reagent in PBS buffer is 2-4 mg / mL.

[0027] Furthermore, in step X1, the mass ratio of the lysozyme to Traut's reagent is 25:1.

[0028] Furthermore, the present invention also provides a method for preparing the long-lasting antibacterial deodorant for sewage, comprising the following steps:

[0029] S1. Add p-tert-amylphenol, methyl parahydroxybenzoate, and activated carbon to anhydrous ethanol, stir at 200-300 rpm for 4-6 hours, and dry under vacuum at 40°C to obtain a mixture.

[0030] S2. Add the modified polymer and modified sustained-release microspheres to a PBS buffer solution at pH 7.4, stir at 60°C and 600 rpm for 1 h, cool, add the modified lysozyme, stir at 100-200 rpm for 2 h, add the mixture obtained in step S1 and 1,2-benzisothiazolin-3-one, stir at 100-150 rpm for 2 h, let stand for 30 min, extrude into granules, shape, and dry to obtain a long-lasting antibacterial deodorant for sewage.

[0031] Furthermore, in step S1, the mass concentration of the activated carbon in anhydrous ethanol is 50-60 mg / mL.

[0032] Furthermore, in step S2, the mass concentration of the modified polymer in PBS buffer is 0.16-0.2 g / mL.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] The present invention provides a long-lasting antibacterial and deodorizing agent for sewage and a preparation method thereof. The agent utilizes antibacterial ingredients such as 1,2-benzisothiazolin-3-one and p-tert-amylphenol, along with activated carbon for odor absorption. Modified sustained-release microspheres and polymers are combined with modified lysozyme and plant extracts to enhance bactericidal efficacy and biocompatibility. The multi-components work synergistically to provide long-lasting antibacterial effects in sewage. The agent also removes odors such as ammonia and hydrogen sulfide through adsorption and neutralization reactions. The agent is suitable for deodorizing and sterilizing sewage tanks in equipment such as sweeping robots and floor scrubbers. The agent utilizes 1,2-benzisothiazolin-3-one, p-tert-amylphenol, and methyl parahydroxybenzoate as chemical antibacterial ingredients, which can penetrate bacterial cell membranes, inhibit microbial growth, and effectively kill bacteria. The addition of lysozyme synergizes with the chemical bactericide to enhance the bactericidal rate. Extracts from plants such as Artemisia annua, licorice, and cloves contain antibacterial active ingredients such as flavonoids and terpenoids. The synergistic effects of these components effectively enhance the bacteriostasis rate. Activated carbon deodorizes by physical adsorption, and the active substances in the plant extracts can react with malodorous gases such as hydrogen sulfide. The cross-linked network structure of the modified polymer, modified lysozyme, and modified sustained-release microspheres can capture odor molecules. The present invention loads plant extracts on porous microspheres to promote the sustained release of active ingredients and improve long-term effectiveness. The surface of the porous microspheres loaded with plant extracts is modified with a polydopamine coating to enhance adhesion. The catechol groups of polydopamine are introduced on the surface of the porous microspheres to form borate ester bonds with the phenylboronic acid groups in the modified polymer. Through cross-linking of the microsphere-polymer network, the antibacterial active substances are encapsulated, the burst release is reduced, and the long-term effectiveness is improved. The modified sustained-release microspheres and the modified polymer form a "double controlled release channel". When bacterial proliferation causes a decrease in local pH, the borate ester bond breaks, releasing more antibacterial ingredients. The reduced bacterial count can maintain long-term sustained release. The present invention adopts AM, AAPBA containing phenylboronic acid groups and NAPA containing amino groups as reaction monomers, polymerizes to form modified polymers, introduces phenylboronic acid groups and amino groups on the molecular chain, and the phenylboronic acid groups can target and bind to bacterial surface polysaccharides (such as lipopolysaccharide) in their free state in addition to forming boronate bonds, thereby destroying the integrity of the cell membrane; at the same time, the catechol group of the polydopamine coating has antioxidant activity and inhibits the formation of bacterial biofilms. The amino group is modified by SMMP, and a maleimide group is introduced, which is covalently bonded to the sulfhydryl group of the modified lysozyme to fix the lysozyme to the polymer network, thereby improving stability and reducing enzyme inactivation. The present invention modifies lysozyme by Traut's reagent, and the sulfhydryl group on the lysine residue of the lysozyme is modified. The modified lysozyme is fixed to the polymer through a sulfhydryl-maleimide bond. The porous structure of the activated carbon can not only adsorb odor molecules (such as ammonia and hydrogen sulfide) in sewage, but also can serve as a physical carrier to adsorb chemical bactericides, slowly release the adsorbed drugs, and prolong the antibacterial effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only for the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0036] Figure 1 The antibacterial effects of the antibacterial deodorants described in Example 1 and Comparative Examples 1-3 of the present invention;

[0037] Figure 2 The deodorizing effects of the antibacterial deodorants of Examples 1-3 and Comparative Examples 1-3 of the present invention;

[0038] Figure 3 The long-lasting antibacterial effect of the antibacterial deodorant described in Example 2 and Comparative Examples 1-3 of the present invention;

[0039] Figure 4 This is a scanning electron micrograph of the modified sustained-release microspheres described in Example 3 of the present invention;

[0040] Figure 5 This is a scanning electron microscope image of the antibacterial deodorant described in Example 1 of the present invention. DETAILED DESCRIPTION

[0041] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with specific examples, but the present invention is not limited to the following examples. It should be noted that, unless otherwise specified, the chemical reagents involved in the present invention were purchased through commercial channels.

[0042] Example 1: A long-lasting antibacterial deodorant for sewage, prepared by preparing raw materials including the following components in parts by weight: 0.3 parts of 1,2-benzisothiazolin-3-one, 0.5 parts of tert-amylphenol, 0.2 parts of methyl parahydroxybenzoate, 6 parts of activated carbon, 6 parts of modified sustained-release microspheres, 12 parts of modified polymers, and 3 parts of modified lysozyme.

[0043] The raw materials for preparing the modified sustained-release microspheres include the following components in parts by weight: 4 parts of Artemisia annua, 3 parts of licorice, 1.5 parts of jasmine, 2 parts of ginkgo leaves, 3 parts of cloves, 1.5 parts of echinacea, 3 parts of dodecylamine (DDA), 7.5 parts of calcium nitrate tetrahydrate (CNT), and 10 parts of dopamine hydrochloride (DA).

[0044] The preparation method of modified sustained-release microspheres comprises the following steps:

[0045] L1. 4 g of Artemisia annua, 3 g of licorice, 1.5 g of jasmine, 2 g of ginkgo biloba, 3 g of cloves, and 1.5 g of Echinacea were dried, ground, and passed through a 60-mesh sieve. The mixture was added to deionized water at a ratio of 1 g:15 mL. The mixture was sonicated at 80 Hz for 30 min, filtered, and the filtrate was concentrated under reduced pressure to 20% of its original volume and freeze-dried to obtain the plant extract.

[0046] L2. Mix 18.72 mL of deionized water and 60 mL of anhydrous ethanol, add 3 g of DDA, and dissolve with stirring at 40°C. Then, at 30-min intervals, add 12 mL of tetraethyl orthosilicate (TEOS), 1.2 mL of triethyl phosphate (TEP), and 7.5 g of CNT dissolved in 15 mL of deionized water. Stir and react for 3 h. Let stand for 22 h, then centrifuge at 10,000 rpm for 15 min. Wash the precipitate with deionized water and ethanol, freeze-dry, calcine at 650°C for 3 h, cool naturally, and grind into a fine powder to obtain porous microspheres.

[0047] L3. Dissolve 10 g of DA at 10 mg / mL in 1000 mL of 10 mM Tris-HCl buffer (pH 8.5) to obtain a DA solution. Add the plant extract and porous microspheres to deionized water. After ultrasonic dispersion, stir at room temperature for 24 h, centrifuge at 10,000 rpm for 15 min, dry the precipitate at 50°C, add it to the DA solution, stir at 200 rpm for 24 h, and centrifuge. Wash the precipitate with deionized water and ethanol, and dry to obtain modified sustained-release microspheres.

[0048] The raw materials for preparing the modified polymer include the following components in parts by weight: 1.5 parts of 3-acrylamidophenylboronic acid (AAPBA), 6 parts of acrylamide (AM), 2.5 parts of N-(3-aminopropyl)acrylamide (NAPA), and 1.8 parts of 3-hydroxysuccinimide maleimidopropionate (SMMP).

[0049] The preparation method of the modified polymer comprises the following steps:

[0050] V1. Add 6 g of AM, 1.5 g of AAPBA, and 2.5 g of NAPA to 300 mL of DMSO solution and stir thoroughly. Vacuum-freeze-thaw-degassed the mixture twice. Add 36 mg of AIBN, seal the container, and vacuum-freeze-thaw-degassed the mixture again. React at 60°C for 12 h. Cool to room temperature, add 10 volumes of ethanol, filter, wash the filter cake with ethanol, and freeze-dry.

[0051] V2. The product obtained in step V1 was added to 900 mL of PBS buffer (pH 7.4) and mixed. 1.8 g of SMMP was added and stirred at 300 rpm for 4 h at room temperature. The product was dialyzed against deionized water (MWCO 7 kDa) for 24 h and freeze-dried to obtain a modified polymer.

[0052] The preparation method of modified lysozyme comprises the following steps:

[0053] X1. Dissolve 2 g of lysozyme in 1000 mL of PBS buffer containing 4 mM EDTA, pH 8.0, to obtain a lysozyme solution. Add 0.08 g of 2-iminothiolane hydrochloride (Traut's reagent) to 20 mL of PBS buffer containing 4 mM EDTA, pH 8.0, to obtain a Traut's reagent solution.

[0054] X2. Mix the Traut's reagent solution obtained in step X1 with the lysozyme solution. Aerate the reaction system with nitrogen and protect from light. Stir at 150 rpm for 2 h. Dialyze against PBS buffer at 4°C and freeze-dry to obtain the modified lysozyme.

[0055] This embodiment also provides a method for preparing the long-lasting antibacterial deodorant for sewage, comprising the following steps:

[0056] S1. Add 0.5 g of p-tert-amylphenol, 0.2 g of methyl parahydroxybenzoate, and 6 g of activated carbon to 100 mL of anhydrous ethanol. Stir at 300 rpm for 6 h. Dry under vacuum at 40°C to obtain a mixture.

[0057] S2. Add 12 g of the modified polymer and 6 g of the modified sustained-release microspheres to 60 mL of PBS buffer (pH 7.4), stir at 60°C and 600 rpm for 1 h, cool, add 3 g of the modified lysozyme, stir at 200 rpm for 2 h, add the mixture obtained in step S1 and 0.3 g of 1,2-benzisothiazolin-3-one, stir at 150 rpm for 2 h, let stand for 30 min, extrude into granules, shape, and dry to obtain a long-lasting antibacterial deodorant for sewage. The scanning electron microscope image is shown in FIG. Figure 5 shown.

[0058] Example 2: A long-lasting antibacterial deodorant for sewage, prepared by preparing raw materials including the following components in parts by weight: 0.2 parts of 1,2-benzisothiazolin-3-one, 0.3 parts of p-tert-amylphenol, 0.1 parts of methyl parahydroxybenzoate, 4 parts of activated carbon, 4 parts of modified sustained-release microspheres, 8 parts of modified polymers, and 2 parts of modified lysozyme.

[0059] The raw materials for preparing the modified sustained-release microspheres include the following components in parts by weight: 3 parts of Artemisia annua, 2 parts of Licorice, 1 part of Jasmine, 1 part of Ginkgo biloba, 2 parts of Clove, 1 part of Echinacea, 2 parts of Dodecylamine (DDA), 5 parts of Calcium Nitrate Tetrahydrate (CNT), and 6 parts of Dopamine Hydrochloride (DA).

[0060] The preparation method of modified sustained-release microspheres comprises the following steps:

[0061] L1. 3 g of Artemisia annua, 2 g of licorice, 1 g of jasmine, 1 g of ginkgo biloba, 2 g of cloves, and 1 g of Echinacea were dried, ground, and passed through a 60-mesh sieve. The mixture was added to deionized water at a ratio of 1 g:10 mL. The mixture was sonicated at 80 Hz for 20 min, filtered, and the filtrate was concentrated under reduced pressure to 20% of its original volume. The mixture was then freeze-dried to obtain the plant extract.

[0062] L2. Mix 12.5 mL of deionized water and 40 mL of anhydrous ethanol, add 2 parts of DDA, and stir to dissolve at 40°C. Then, at 30-min intervals, add 8 mL of tetraethyl orthosilicate (TEOS), 0.8 mL of triethyl phosphate (TEP), and 5 g of CNT dissolved in 10 mL of deionized water. Stir and react for 3 h. Let stand for 18 h, then centrifuge at 10,000 rpm for 10 min. Wash the precipitate with deionized water and ethanol, freeze-dry, calcine at 650°C for 3 h, cool naturally, and grind into a fine powder to obtain porous microspheres.

[0063] L3. Dissolve 6 g of DA at 10 mg / mL in 600 mL of 10 mM Tris-HCl buffer (pH 8.5) to obtain a DA solution. Add the plant extract and porous microspheres to deionized water. After ultrasonic dispersion, stir at room temperature for 24 h, centrifuge at 10,000 rpm for 10 min, dry the precipitate at 50°C, add it to the DA solution, stir at 150 rpm for 24 h, and centrifuge. Wash the precipitate with deionized water and ethanol, and dry to obtain modified sustained-release microspheres.

[0064] The raw materials for preparing the modified polymer include the following components in parts by weight: 1 part of 3-acrylamidophenylboronic acid (AAPBA), 4 parts of acrylamide (AM), 1.5 parts of N-(3-aminopropyl)acrylamide (NAPA), and 1.2 parts of 3-hydroxysuccinimide maleimidopropionate (SMMP).

[0065] The preparation method of the modified polymer comprises the following steps:

[0066] V1. Add 4 g of AM, 1 g of AAPBA, and 1.5 g of NAPA to 200 mL of DMSO solution and stir thoroughly. Vacuum-freeze-thaw-degassed the mixture twice. Add 20 mg of AIBN, seal the container, vacuum-freeze-thaw-degassed the mixture again, and react at 60°C for 10 h. Cool to room temperature, add 10 volumes of ethanol, filter, wash the filter cake with ethanol, and freeze-dry.

[0067] V2. The product obtained in step V1 was added to 600 mL of PBS buffer (pH 7.4) and mixed. 1.2 g of SMMP was added and stirred at 200 rpm at room temperature for 3 h. The product was dialyzed against deionized water (MWCO 5 kDa) for 24 h and freeze-dried to obtain a modified polymer.

[0068] The preparation method of modified lysozyme comprises the following steps:

[0069] X1. Dissolve 2 g of lysozyme in 1000 mL of PBS buffer containing 4 mM EDTA, pH 8.0, to obtain a lysozyme solution. Add 0.08 g of 2-iminothiolane hydrochloride (Traut's reagent) to 40 mL of PBS buffer containing 4 mM EDTA, pH 8.0, to obtain a Traut's reagent solution.

[0070] X2. Mix the Traut's reagent solution obtained in step X1 with the lysozyme solution. Aerate the reaction system with nitrogen and protect from light. Stir at 150 rpm for 1 h. Dialyze against PBS buffer at 4°C and freeze-dry to obtain the modified lysozyme.

[0071] This embodiment also provides a method for preparing the long-lasting antibacterial deodorant for sewage, comprising the following steps:

[0072] S1. Add 0.3 g of p-tert-amylphenol, 0.1 g of methyl parahydroxybenzoate, and 4 g of activated carbon to 80 mL of anhydrous ethanol. Stir at 200 rpm for 4 h. Dry under vacuum at 40°C to obtain a mixture.

[0073] S2. Add 8 g of the modified polymer and 4 g of the modified sustained-release microspheres to 50 mL of PBS buffer (pH 7.4), stir at 60°C and 600 rpm for 1 h, cool, add 2 g of the modified lysozyme, stir at 100 rpm for 2 h, add the mixture obtained in step S1 and 0.2 g of 1,2-benzisothiazolin-3-one, stir at 100 rpm for 2 h, let stand for 30 min, extrude into granules, shape, and dry to obtain a long-lasting antibacterial deodorant for sewage.

[0074] Example 3: A long-lasting antibacterial deodorant for sewage, prepared by preparing raw materials including the following components in parts by weight: 0.25 parts of 1,2-benzisothiazolin-3-one, 0.4 parts of p-tert-amylphenol, 0.15 parts of methyl parahydroxybenzoate, 5 parts of activated carbon, 5 parts of modified sustained-release microspheres, 10 parts of modified polymers, and 2.5 parts of modified lysozyme.

[0075] The raw materials for preparing the modified sustained-release microspheres include the following components in parts by weight: 3.5 parts of Artemisia annua, 2.5 parts of licorice, 1.2 parts of jasmine, 1.5 parts of ginkgo leaves, 2.5 parts of cloves, 1.2 parts of echinacea, 2.5 parts of dodecylamine (DDA), 6 parts of calcium nitrate tetrahydrate (CNT), and 8 parts of dopamine hydrochloride (DA).

[0076] The preparation method of modified sustained-release microspheres comprises the following steps:

[0077] L1. 3.5 g of Artemisia annua, 2.5 g of licorice, 1.2 g of jasmine, 1.5 g of ginkgo biloba, 2.5 g of cloves, and 1.2 g of Echinacea were dried, ground, and passed through a 60-mesh sieve. The mixture was added to deionized water at a ratio of 1 g:12 mL. The mixture was sonicated at 80 Hz for 25 min, filtered, and the filtrate was concentrated under reduced pressure to 20% of its original volume. The resulting mixture was then freeze-dried to obtain the plant extract.

[0078] L2. Mix 15.625 mL of deionized water and 50 mL of anhydrous ethanol, add 2.5 g of DDA, and dissolve with stirring at 40°C. Then, at 30-min intervals, add 10 mL of tetraethyl orthosilicate (TEOS), 1 mL of triethyl phosphate (TEP), and 6 g of CNT dissolved in 12 mL of deionized water. Stir and react for 3 h. Let stand for 20 h, then centrifuge at 10,000 rpm for 12 min. Wash the precipitate with deionized water and ethanol, freeze-dry, calcinate at 650°C for 3 h, cool naturally, and grind into a fine powder to obtain porous microspheres.

[0079] L3. DA solution was prepared by dissolving 8 g of DA at 10 mg / mL in 800 mL of 10 mM Tris-HCl buffer (pH 8.5). The plant extract and porous microspheres were added to deionized water, and ultrasonically dispersed. The mixture was stirred at room temperature for 24 h, centrifuged at 10,000 rpm for 12 min, and the precipitate was dried at 50°C. The mixture was added to the DA solution, stirred at 180 rpm for 24 h, and centrifuged. The precipitate was washed with deionized water and ethanol, and dried to obtain modified sustained-release microspheres. The scanning electron micrograph is shown in FIG. Figure 4 shown.

[0080] The raw materials for preparing the modified polymer include the following components in parts by weight: 1.2 parts of 3-acrylamidophenylboronic acid (AAPBA), 5 parts of acrylamide (AM), 2 parts of N-(3-aminopropyl)acrylamide (NAPA), and 1.5 parts of 3-hydroxysuccinimide maleimidopropionate (SMMP).

[0081] The preparation method of the modified polymer comprises the following steps:

[0082] V1. Add 5 g of AM, 1.2 g of AAPBA, and 2 g of NAPA to 250 mL of DMSO solution and stir thoroughly. Vacuum-freeze-thaw-degassed the mixture twice. Add 27.5 mg of AIBN, seal the container, and vacuum-freeze-thaw-degassed the mixture again. React at 60°C for 11 h. Cool to room temperature, add 10 times the volume of ethanol, filter, wash the filter cake with ethanol, and freeze-dry.

[0083] V2. The product from step V1 was added to 750 mL of PBS buffer (pH 7.4) and mixed. 1.5 g of SMMP was added and the mixture was stirred at 250 rpm at room temperature for 3.5 h. The mixture was dialyzed against deionized water (MWCO 6 kDa) for 24 h and freeze-dried to obtain a modified polymer.

[0084] The preparation method of modified lysozyme comprises the following steps:

[0085] X1. Dissolve 2 g of lysozyme in 1000 mL of PBS buffer containing 4 mM EDTA, pH 8.0, to obtain a lysozyme solution. Add 0.08 g of 2-iminothiolane hydrochloride (Traut's reagent) to 27 mL of PBS buffer containing 4 mM EDTA, pH 8.0, to obtain a Traut's reagent solution.

[0086] X2. Mix the Traut's reagent solution obtained in step X1 with the lysozyme solution. Aerate the reaction system with nitrogen and protect from light. Stir at 150 rpm for 1.5 h. Dialyze against PBS buffer at 4°C and freeze-dry to obtain the modified lysozyme.

[0087] This embodiment also provides a method for preparing the long-lasting antibacterial deodorant for sewage, comprising the following steps:

[0088] S1. Add 0.4 g of p-tert-amylphenol, 0.15 g of methyl parahydroxybenzoate, and 5 g of activated carbon to 90 mL of anhydrous ethanol. Stir at 250 rpm for 5 h. Dry under vacuum at 40°C to obtain a mixture.

[0089] S2. Add 10 g of the modified polymer and 5 g of the modified sustained-release microspheres to 60 mL of PBS buffer (pH 7.4), stir at 60°C and 600 rpm for 1 h, cool, add 2.5 g of the modified lysozyme, stir at 150 rpm for 2 h, add the mixture obtained in step S1 and 0.25 g of 1,2-benzisothiazolin-3-one, stir at 120 rpm for 2 h, let stand for 30 min, extrude into granules, shape, and dry to obtain a long-lasting antibacterial deodorant for sewage.

[0090] The only difference between Comparative Example 1 and Example 1 is that no modified sustained-release microspheres are added.

[0091] The only difference between Comparative Example 2 and Example 1 is that no modified polymer is added.

[0092] The only difference between Comparative Example 3 and Example 1 is that no modified lysozyme is added.

[0093] Experimental Example 1: Staphylococcus aureus and Escherichia coli were used to verify the antibacterial performance of the antibacterial deodorant. 8 CFU / mL of bacterial solution was mixed with 50 μL of LB culture medium, and 0.05 g of the antibacterial deodorant of Example 1 and Comparative Examples 1-3 was added as the experimental group. The group without antibacterial deodorant was used as the control group. The culture was placed in a constant temperature shaker at 37°C and 120 rpm for 12 h. The final bacterial solution was diluted with 0.9% physiological saline to a concentration of 1×10 3 CFU / mL, 100 μL of diluted bacterial solution was evenly spread on the agar plate medium, placed in a constant temperature incubator at 37℃ for 12 h, the number of colonies was counted, and the sterilization rate was calculated. Sterilization rate (%) = [(number of colonies in the control group - number of colonies in the experimental group) / number of colonies in the control group] × 100%. The results are as follows Figure 1 shown.

[0094] Figure 1 The results showed that the bactericidal rate of Example 1 group reached more than 99%, which was higher than that of Comparative Examples 1 to 3, and had better bactericidal effect, indicating that the antibacterial deodorant of the present invention can effectively kill bacteria, is suitable for sewage, and the components synergize to enhance the effect.

[0095] Experimental Example 2: Prepare standard pollutants: 10 mL of milk, 5 mL of soy sauce, 0.5 g of hair, and 1 g of indoor dust. Take 20 g of the antibacterial deodorant of Examples 1-3 and Comparative Examples 1-3 and add them to a container containing 4 L of tap water and pollutants. After the container is sealed and left to stand at room temperature for 7 days, the odor is detected using the fan smelling method (i.e., place the sample about 20 cm below the nose, fan it gently with your hand, and smell it to avoid inhaling a large amount of gas). The odor detection standard is shown in Table 1. The test results are shown in Table 1. Figure 2 shown.

[0096] Table 1:

[0097]

[0098] Figure 2 The results showed that the odor intensity of Examples 1-3 was lower than that of Comparative Examples 1-3. Comparative Example 1 did not add modified sustained-release microspheres, lacked plant extracts, and could not neutralize odor molecules; Comparative Example 2 did not add modified polymers, and the active ingredients were easily ineffective after sudden release, and the polymer network did not fix the lysozyme, which was easily inactivated, accelerated the reproduction of microorganisms, and produced foul-smelling gases; Comparative Example 3 did not add modified lysozyme, and the odor removal effect was reduced. The activated carbon and plant extracts could only partially adsorb / neutralize the odor.

[0099] Experimental Example 3: According to the method of Experimental Example 1, at the end of each antibacterial cycle (3rd, 5th, 7th, and 15th days), the bacterial solution was added again according to the above process, the plates were coated, and the sterilization rate (%) was calculated. The results are as follows: Figure 3 shown.

[0100] Figure 3 The results showed that the bactericidal rate of Example 2 was better than that of Comparative Examples 1-3, and the long-term antibacterial effect was better than that of Comparative Examples 1-3. The modified sustained-release microspheres, modified polymers, and modified lysozyme worked synergistically. Comparative Example 1 did not add modified sustained-release microspheres, and the plant antibacterial components could not be sustained-released; Comparative Example 2 did not add modified polymers, and a microsphere-polymer dynamic network could not be formed, lysozyme could not be fixed, and the enzyme was easily inactivated; Comparative Example 3 did not add modified lysozyme, and lacked the bactericidal activity of lysozyme.

[0101] 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. Within the scope 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.

Claims

1. A long-lasting antibacterial deodorant for sewage, characterized in that: The preparation raw materials include the following components in parts by weight: 0.2-0.3 parts of 1,2-benzisothiazolin-3-one, 0.3-0.5 parts of p-tert-amylphenol, 0.1-0.2 parts of methyl parahydroxybenzoate, 4-6 parts of activated carbon, 4-6 parts of modified sustained-release microspheres, 8-12 parts of modified polymer, and 2-3 parts of modified lysozyme; The preparation method of the modified sustained-release microspheres comprises the following steps: L1. By weight, 3-4 parts of Artemisia annua, 2-3 parts of licorice, 1-1.5 parts of jasmine, 1-2 parts of ginkgo biloba, 2-3 parts of cloves, 1-1.5 parts of Echinacea were dried and crushed, ultrasonically extracted with water, concentrated and freeze-dried to obtain a plant extract; L2 was mixed with water and ethanol, 2-3 parts of dodecylamine was added to dissolve, tetraethyl orthosilicate, triethyl phosphate, 5-7.5 parts of calcium nitrate tetrahydrate was added, stirred, allowed to stand, freeze-dried, and calcined to obtain porous microspheres; L3 6-10 parts of dopamine hydrochloride were dissolved in Tris-HCl buffer to obtain a dopamine hydrochloride solution, the plant extract and porous microspheres were added to water, sonicated, stirred, centrifuged, added to the dopamine hydrochloride solution, stirred to obtain modified sustained-release microspheres; The preparation method of the modified polymer comprises the following steps: V1. Add 4-6 parts of acrylamide, 1-1.5 parts of 3-acrylamidophenylboronic acid, and 1.5-2.5 parts of N-(3-aminopropyl)acrylamide to a DMSO solution by weight, degas, add azobisisobutyronitrile, degas, react, add ethanol, filter, wash, and freeze-dry; V2 The product obtained in step V1 was added to PBS buffer, 1.2-1.8 parts of 3-maleimido propionic acid hydroxysuccinimide ester was added, stirred, dialyzed, and lyophilized to obtain a modified polymer; The preparation method of the modified lysozyme comprises the following steps: X1. Take lysozyme and add it to PBS buffer to obtain a lysozyme solution. Add 2-iminothiolane hydrochloride to PBS buffer to obtain a 2-iminothiolane hydrochloride solution. X2. The 2-iminothiolane hydrochloride solution obtained in step X1 was mixed with the lysozyme solution, filled with nitrogen, protected from light, stirred, dialyzed, and lyophilized to obtain modified lysozyme.

2. The long-lasting antibacterial deodorant for sewage according to claim 1, characterized in that: In step L2, the volume ratio of water to ethanol is 5:16; the usage ratio of dodecylamine to anhydrous ethanol is 1 g:20 mL; and the volume ratio of tetraethyl orthosilicate, triethyl phosphate, and anhydrous ethanol is 10:1:

50.

3. The long-lasting antibacterial deodorant for sewage according to claim 2, characterized in that: In step V2, the mass concentration of the 3-maleimidopropionic acid hydroxysuccinimide ester in the PBS buffer solution is 2 mg / mL.

4. The long-lasting antibacterial deodorant for sewage according to claim 3, characterized in that: In step X1, the mass ratio of lysozyme to 2-iminothiolane hydrochloride is 25:

1.

5. A method for preparing the long-lasting antibacterial deodorant for sewage according to any one of claims 1 to 4, characterized in that: The following steps are involved: S1. p-tert-amylphenol, methyl parahydroxybenzoate and activated carbon were added to anhydrous ethanol, stirred, and dried to obtain a mixture; S2. Add the modified polymer and modified sustained-release microspheres to PBS buffer solution, stir, add modified lysozyme, stir, add the mixture obtained in step S1 and 1,2-benzisothiazolin-3-one, stir, let stand, granulate, shape, and dry to obtain a long-lasting antibacterial deodorant for sewage.

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