Microbial peculiar smell removing agent suitable for environment and preparation method of microbial peculiar smell removing agent

Abstract

The invention discloses a microbial odor remover suitable for environment and a preparation method thereof, and relates to the technical field of daily chemical products. The preparation method comprises the following steps: adding pediococcus acidilactici and bacillus velezensis into a citric acid-disodium hydrogen phosphate buffer solution according to a ratio, standing and activating, adding bromelain, lipase, rosmarinic acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA (Ethylene Diamine Tetraacetic Acid) mixture, homogenizing, adding konjac glucomannan-chitosan gel and a trehalose-glycerol compound according to a ratio, stirring, filtering, and drying to obtain a finished product. And subpackaging in a dark place to prepare the microbial odor remover suitable for the environment. Through the synergistic effect of microorganisms, enzymes and a natural deodorization medium, a multi-stage peculiar smell elimination system is constructed, the dual functions of immediate neutralization and continuous bacteriostasis are achieved, the application scene is wide, corrosion and residues to base materials such as fabric and plastic are avoided, and the concentration of harmful gas can be more effectively controlled and pathogenic microorganisms can be inhibited in large-scale breeding application.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of daily chemical products, in particular to a microbial odor eliminator suitable for the environment and a preparation method thereof. BACKGROUND

[0002] Odor problems are widespread in pet products, home environments and large-scale breeding fields, mainly due to volatile malodorous substances such as ammonia, hydrogen sulfide, indole and other volatile malodorous substances produced by protein corruption, fat oxidation and microbial metabolism. The existing deodorization technology has obvious limitations: chemical masking agents only cover the odor temporarily and may cause secondary pollution; physical adsorption materials are easy to release pollutants after saturation; traditional microbial preparations are difficult to continuously exert their efficacy in complex substrates due to low activity of bacterial strains and poor environmental adaptability. Especially for high organic load scenes such as manure in breeding farms and rotten kitchen waste, conventional products often have problems such as inactivation of bacterial flora, sudden decrease of enzymatic efficiency or incompatibility of ingredients, resulting in incomplete deodorization and short effective period. In recent years, although some researches have tried to use a composite bacteria-enzyme synergistic system, the core problems have not been solved, i.e. the survival of microorganisms in acidic / alkaline environments, the catalytic stability of enzymes in oil interfaces, and the targeted degradation ability of stubborn sulfur and nitrogen compounds. Therefore, it is urgent to develop an environmentally friendly microbial odor eliminator that can penetrate the barrier of organic pollutants, has instant deodorization and long-term bacteriostatic function, and has no toxic side effects on animals and plants. SUMMARY

[0003] The present application aims to provide a microbial odor eliminator suitable for the environment and a preparation method thereof, and its preparation method and application, to solve the problems raised in the background art.

[0004] To achieve the above-mentioned purpose, the present application provides the following technical solutions:

[0005] A microbial odor eliminator suitable for the environment, comprising the following raw material components by weight fraction:

[0006] Pediococcus acidilactici 15-25 parts;

[0007] Bacillus velezensis 15-25 parts;

[0008] Papain 5-10 parts;

[0009] Lipase 3-8 parts;

[0010] Rosmarinic acid nanoemulsion 10-20 parts;

[0011] Tea polyphenol 5-12 parts;

[0012] Citrus polyphenol-EDTA mixture 8-15 parts;

[0013] Konjac glucomannan-chitosan gel 10-18 parts;

[0014] Citric acid-disodium hydrogen phosphate buffer 20-30 parts;

[0015] Trehalose-glycerol complex 8-15 parts.

[0016] Further, the Pediococcus acidilactici needs to be microencapsulated with sodium alginate, and the specific operation is as follows:

[0017] The Pediococcus acidilactici is inoculated in MRS liquid medium with pH of 6.2, and fermented at 37°C for 24h to obtain a bacterial concentration of ≥1x10 9 CFU / mL; the bacterial solution is mixed with a sodium alginate solution with a mass fraction of 2.5%, and dropped into a CaCl2 solution with a mass fraction of 2% through a microcapsule generator, and the microcapsules are collected after solidification for 15min and freeze-dried for storage;

[0018] The volume ratio of the bacterial solution to the sodium alginate solution is 1:3;

[0019] The voltage of the microcapsule generator is 6-10kV, and the needle diameter is 0.5mm.

[0020] It should be noted that microencapsulation with sodium alginate can improve the survival rate and slow-release efficiency of the bacterial body. The microcapsule structure resists the strong acid environment of vomit / excrement, prevents the bacterial body from being inactivated too early, and the enzymes produced by the Pediococcus acidilactici in the environment gradually hydrolyze the capsule wall network, realizing the progressive colonization of Pediococcus acidilactici on the surface of contaminated organic matter and releasing Pediococcus acidilactici to directionally decompose nitrogen-containing foul-smelling molecules such as uric acid and indole, and convert them into odorless lactic acid.

[0021] Further, the Bacillus velezensis is coated with a polydopamine coating on the surface, and the specific operation is as follows:

[0022] The Bacillus velezensis is cultured in GYS medium containing 0.1% mass fraction of MnSO4 at 48-52°C for 36-48h to obtain a spore suspension; the spore suspension is mixed with a polydopamine solution with a concentration of 0.1-0.3mg / mL, the pH is adjusted to 8.5, and the reaction is carried out for 3-6h, and the Bacillus velezensis with polydopamine adhesion coating on the surface is obtained after centrifugal washing;

[0023] The volume ratio of the spore suspension to the polydopamine solution is 1:(3-5).

[0024] It should be noted that the polydopamine coating enhances the initial adhesion of bacillus velezensis spores on the surface of hydrophobic organic pollutants (oil, protein particles) through the adhesion characteristics of catechol / quinone groups. After spore germination, the bacteria secrete extracellular proteases to degrade surrounding pollutants (such as putrefactive proteins) to generate small molecule peptides / amino acids for bacterial growth, while secreting amine oxidase to decompose putrescine / cadaverine into aldehyde and hydrogen peroxide, and finally metabolized into harmless substances. Polydopamine does not directly participate in the decomposition of pollutants, but indirectly enhances the biodegradation process by improving the retention efficiency of the bacteria on the pollution interface.

[0025] Further, the bromelain needs to be encapsulated by ZIF-8 metal framework, and the specific operation is as follows:

[0026] 0.5 mmol (0.149 g) Zn(NO3)2·6H2O was dissolved in 5 mL methanol solution in an ice water bath at 2-6℃, the pH was adjusted to 6-8, 20 mg of bromelain was added, 0.4 mmol (0.033 g) 2-methyl imidazole was added in 10 mL methanol solution, stirred for 8-10 min, and then centrifuged to collect the precipitate, washed and dried to obtain ZIF-8 encapsulated bromelain;

[0027] The methanol solution is prepared by mixing methanol and pure water at a volume ratio of 3:7;

[0028] The amount ratio of Zn(NO3)2·6H2O, methanol, bromelain, 2-methyl imidazole and methanol solution is (14.5-15.3) g:500 mL:(1.8-2.2) g:(3-3.6) g:1000 mL.

[0029] It should be noted that bromelain can target and decompose protein-based malodor sources, destroy microbial biofilms, and block the enzymatic reaction of microorganisms converting tryptophan into skatole, thereby reducing one of the sources of malodor from the source. Zn 2+ is combined with deprotonated 2-methyl imidazole through coordination bond, Zn 2+ is attracted by the negative charge on the enzyme surface, bromelain acts as a template to provide heterogeneous nucleation sites, and ZIF-8 grows epitaxially on the enzyme surface to form a ZIF-8 framework that protects the enzyme from inactivation in acidic environments (pH <4).

[0030] Further, the lipase needs to be modified by polyethylene glycol-phospholipid, and the specific operation is as follows:

[0031] Mix the lipase with 1,2-distearyl-phosphatidylethanolamine-polyethylene glycol in a phosphate buffer at pH 7.4, and oscillate the reaction at 35℃ for 3h. Remove the free phospholipid by ultrafiltration, and freeze-dry to obtain the polyethylene glycol-phospholipid modified lipase;

[0032] The molecular weight of the polyethylene glycol in the 1,2-distearoyl-phosphatidyl ethanolamine-polyethylene glycol is 2000.

[0033] The ratio of the lipase, 1,2-distearoyl-phosphatidyl ethanolamine-polyethylene glycol and phosphate buffer is 1g:(18-22)g:(200-300)mL.

[0034] It should be noted that the phospholipid layer can enhance the catalytic efficiency of the lipase at the oil-water interface. The lipase has an affinity for the oil-water interface and can catalyze the hydrolysis of water-insoluble lipid substances at a high rate on the oil-water interface, acting on the hydrophilic-hydrophobic interface layer of the system. The hydrophobic phospholipid tail of the 1,2-distearoyl-phosphatidyl ethanolamine-polyethylene glycol inserts into the hydrophobic pocket of the lipase, and the hydrophilic chain of the polyethylene glycol extends outward, forming a "hydrophobic insertion anchoring-hydrophilic shielding" complex structure. Surface modification allows the lipase to be oriented and arranged at the oil-water interface, increasing the catalytic efficiency and allowing rapid hydrolysis of short-chain fatty acid esters produced by kitchen waste spoilage. The long chain of polyethylene glycol can prevent the aggregation of lipase in an organic solvent environment (such as urine), maintain catalytic activity, and delay the conformational collapse of lipase in a high-temperature environment (45°C), ensuring stability in high-temperature environments.

[0035] Further, the preparation steps of the rosemary nanoemulsion are as follows:

[0036] Mix rosemary acid, Tween 80 and isopropyl myristate and heat to 60-70°C; slowly add 60-70°C pure water, and process with a homogenizer at 10000-12000r / min for 8-12min; cool to room temperature and sterilize through a 0.22μm filter membrane;

[0037] The mass ratio of the rosemary acid, Tween 80, isopropyl myristate and pure water is 1:4:1:54.

[0038] Further, the preparation steps of the citrus polyphenol-EDTA mixture are as follows:

[0039] Dissolve the citrus polyphenol and disodium ethylenediaminetetraacetate in an ethanol solution, stir and react at 55-65°C for 6-8h, rotary evaporate at 40-50°C, and freeze-dry to obtain the citrus polyphenol-EDTA mixture;

[0040] The total phenol content of the citrus polyphenol is greater than or equal to 60%;

[0041] The mass ratio of the citrus polyphenol and disodium ethylenediaminetetraacetate is (1.6-2):1;

[0042] The ethanol solution is prepared from ethanol and pure water in a volume ratio of 1:1;

[0043] The ratio of the citrus polyphenol and the ethanol solution is 1g:(20-30)mL.

[0044] It should be noted that the citrus polyphenol-EDTA mixture can simultaneously remove ammonia and metal ion odorants. The ortho-diphenol hydroxyl group of citrus polyphenol and the carboxyl group of disodium ethylenediaminetetraacetate (EDTA) can coordinate with metal ions in feces and urine to form a mixed ligand complex, block the catalytic production of thiol-based malodors by metal ions, disrupt the microbial iron uptake pathway, and inhibit the proliferation of common spoilage bacteria (such as Clostridium). In addition, the phenolic hydroxyl proton can combine with NH3 to form ammonium salt, neutralizing ammonia gas.

[0045] Further, the preparation steps of the konjac glucomannan-chitosan gel are as follows:

[0046] Mix konjac glucomannan and chitosan, add 1% acetic acid solution to the mixture at 30-40°C, adjust the pH to 5-5.5, stir for 15-30 min, and obtain konjac glucomannan-chitosan gel;

[0047] The degree of deacetylation of chitosan is ≥90%;

[0048] The amount ratio of konjac glucomannan, chitosan and acetic acid solution is 1g:(1-1.2)g:(30-50)mL.

[0049] Further, the pH of the citric acid-disodium hydrogen phosphate buffer is 5.8, and the concentration of the buffer system is 0.1 mol / L, wherein the mass ratio of citric acid, disodium hydrogen phosphate and pure water is (9.5-10.5):(7.1-7.8):800.

[0050] Further, the trehalose-glycerol complex is prepared by melting and blending trehalose and glycerol at 55-65°C and then quenching;

[0051] The mass ratio of trehalose to glycerol is 1:1.

[0052] Further, when applied to poultry and livestock farms, nitrifying bacteria can be added to the raw material components.

[0053] A preparation method of a microbial odor-removing agent suitable for the environment, comprising the following steps:

[0054] Step one: according to the proportion, add Pediococcus acidilactici and Bacillus velezensis to a citric acid-disodium hydrogen phosphate buffer containing 0.5% sucrose, and activate at 30-40°C for 2-4h;

[0055] Step two: according to the proportion, add bromelain, lipase, rosemary acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture to the system obtained in step one, and homogenize at 28-32°C for 3-8min;

[0056] Step three, according to the proportion, konjac glucoside-chitosan gel and trehalose-glycerol complex are added, stirred, and light-protected to prepare the environment-friendly microbial odor removal agent.

[0057] The use method of the environment-friendly microbial odor removal agent is: after being diluted with pure water to a required proportion, the environment-friendly microbial odor removal agent is sprayed for use.

[0058] Compared with the prior art, the present application has the following beneficial effects:

[0059] The present application builds a multi-stage odor removal system through the synergistic effect of microorganisms, enzymes and natural deodorizing media. The core bacterial population (Pediococcus acidilactici and Bacillus velezensis) maintains high activity and directionally degrades organic pollutants in complex environments after being embedded or surface modified, solving the problem of poor environmental tolerance of traditional probiotics. The enzyme system (bromelain and lipase) is encapsulated or interface modified, and the catalytic efficiency in extreme pH and oil environments is improved, realizing efficient decomposition of protein, fat and other odor-causing molecules. The deodorizing medium (rosemary acid nanoemulsion, tea polyphenol, and citrus polyphenol-EDTA mixture) can eliminate sulfur-containing, nitrogen-containing volatile substances and fecal odor molecules, and accurately capture gaseous odor molecules and metal ions.

[0060] The present application has the dual functions of instant neutralization and continuous bacteriostasis, is suitable for a wide range of applications, has no corrosion and no residue on fabrics, plastics and other substrates, can effectively control the concentration of harmful gases and inhibit the growth of pathogenic microorganisms in large-scale breeding applications, and has a complete mechanism, which can solve the problems of single effect and short duration of traditional deodorizing products. DETAILED DESCRIPTION

[0061] The technical solutions in the embodiments of the present application will be described below in a clear and complete manner. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0062] Preparation Example 1

[0063] Pediococcus acidilactici is from Beijing Yujin Technology Co., Ltd., and the strain number is CICC 10346.

[0064] The preparation steps of sodium alginate microencapsulated Pediococcus acidilactici are as follows:

[0065] Pediococcus acidilactici is inoculated in MRS liquid medium with a pH of 6.2, and fermented at 37 DEG C for 24 hours until the bacterial concentration is greater than or equal to 1x10 9CFU / mL; the bacterial solution was mixed with a 2.5% sodium alginate solution at a volume ratio of 1:3, dropped into a 2% CaCl2 solution through a microcapsule generator (voltage 8 kV, needle diameter 0.5 mm), and the microcapsules were collected after being solidified for 15 min and stored after freeze-drying.

[0066] Preparation Example 2

[0067] Bacillus velezensis was from Beijing Yujin Technology Co., Ltd., and the strain number was CICC 22799.

[0068] Preparation steps of Bacillus velezensis with a polydopamine coating on the surface:

[0069] Bacillus velezensis was cultured at 50°C for 48 h in a GYS medium containing 0.1% MnSO4 to obtain a spore suspension; the spore suspension was mixed with a 0.2 mg / mL polydopamine solution at a volume ratio of 1:4, the pH was adjusted to 8.5, and the reaction was performed for 4 h; after centrifugal washing, Bacillus velezensis with a polydopamine adhesion coating on the surface was obtained.

[0070] Preparation Example 3

[0071] Preparation steps of ZIF-8 metal framework encapsulated bromelain:

[0072] 0.149 g Zn(NO3)2·6H2O was dissolved in 5 mL of a methanol solution in an ice water bath at 4°C, the pH was adjusted to 8, 20 mg of bromelain was added, 10 mL of a methanol solution containing 33 mg of 2-methylimidazole was added, stirring was performed for 10 min, and the precipitate was collected by centrifugation after standing for 3 min; after washing and drying, ZIF-8 encapsulated bromelain was obtained; the methanol solution was prepared by mixing methanol and pure water at a volume ratio of 3:7.

[0073] Preparation Example 4

[0074] Preparation steps of polyethylene glycol-phospholipid modified lipase:

[0075] 1 g of lipase was mixed with 20 g of 1,2-distearyl-phosphatidylethanolamine-polyethylene glycol (polyethylene glycol molecular weight 2000) in 250 mL of a phosphate buffer with a pH of 7.4, and the reaction was performed by oscillation at 35°C for 3 h; free phospholipids were removed by ultrafiltration, and polyethylene glycol-phospholipid modified lipase was obtained by freeze-drying.

[0076] Preparation Example 5

[0077] The preparation steps of rosemary nanoemulsion are as follows:

[0078] Mixing 1 g rosemary acid, 4 g Tween 80, 1 g isopropyl myristate, heating to 65℃; slowly add 54 g 65℃ pure water, homogenizer 12000r / min for 10 min; after cooling to room temperature, sterilize through 0.22μm filter membrane;

[0079] Preparation Example 6

[0080] The preparation steps of the citrus polyphenol-EDTA mixture are as follows:

[0081] Dissolve 1.82 g citrus polyphenol (total phenol content 60%) and 1 g ethylenediaminetetraacetic acid disodium salt in 25 mL ethanol solution, stir at 60℃ for 6 h, rotary evaporation at 45℃, freeze-drying to obtain a citrus polyphenol-EDTA mixture;

[0082] The ethanol solution is prepared by mixing ethanol and pure water in a volume ratio of 1:1;

[0083] Preparation Example 7

[0084] The preparation steps of the konjac glucomannan-chitosan gel are as follows:

[0085] Mix 1 g konjac glucomannan and 1.05 g chitosan (degree of deacetylation ≥90%) and add to 40 mL acetic acid solution with an acetic acid mass fraction of 1% at 35℃, adjust the pH to 5.5, stir for 25 min to obtain a konjac glucomannan-chitosan gel;

[0086] Preparation Example 8

[0087] Add 9.82 g citric acid and 7.56 g disodium hydrogen phosphate to 800 g pure water, adjust the pH to 5.8 to obtain a citric acid-disodium hydrogen phosphate buffer solution.

[0088] Preparation Example 9

[0089] The preparation steps of the trehalose-glycerol complex are as follows:

[0090] Mix 1 g trehalose and 1 g glycerol at 60℃, then quench to obtain a trehalose-glycerol complex.

[0091] Example 1

[0092] A microbial odor remover suitable for the environment, which comprises the following raw material components in parts by weight:

[0093] Pediococcus acidilactici 20 parts;

[0094] Bacillus velezensis 20 parts;

[0095] Papain 8 parts;

[0096] Lipase 5 parts;

[0097] Rosemary acid nanoemulsion 15 parts;

[0098] Tea polyphenol 8 parts;

[0099] Citrus polyphenol-EDTA mixture 12 parts;

[0100] Konjac glucomannan-chitosan gel 14 parts;

[0101] Citric acid-disodium hydrogen phosphate buffer 25 parts;

[0102] Trehalose-glycerol complex 12 parts.

[0103] A preparation method of a microbial odor cleaner suitable for the environment, comprising the following steps:

[0104] Step one, according to the proportion, lactococcus lactis and bacillus velezensis are added into citric acid-disodium hydrogen phosphate buffer containing 0.5% mass fraction of sucrose, and activated at 35℃ for 2h;

[0105] Step two, according to the proportion, bromelain, lipase, rosemary acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture are added into the system obtained in step one, and homogenized at 30℃ for 5min;

[0106] Step three, according to the proportion, konjac glucomannan-chitosan gel and trehalose-glycerol complex are added, stirred, and packaged in the dark to obtain a microbial odor cleaner suitable for the environment.

[0107] The lactococcus lactis, bacillus velezensis, bromelain, lipase, rosemary acid nanoemulsion, citrus polyphenol-EDTA mixture, konjac glucomannan-chitosan gel, citric acid-disodium hydrogen phosphate buffer and trehalose-glycerol complex used in this example are all prepared by preparation examples 1-9, and other examples are the same.

[0108] Example 2

[0109] A microbial odor cleaner suitable for the environment, comprising the following raw material components by weight fraction:

[0110] Lactococcus lactis 15 parts;

[0111] Bacillus velezensis 15 parts;

[0112] Bromelain 5 parts;

[0113] Lipase 3 parts;

[0114] Rosemary acid nanoemulsion 10 parts;

[0115] Tea polyphenol 5 parts;

[0116] Citrus polyphenol-EDTA mixture 8 parts;

[0117] Konjac glucomannan-chitosan gel 10 parts;

[0118] Citric acid-disodium hydrogen phosphate buffer 20 parts;

[0119] Trehalose-glycerol complex 8 parts.

[0120] A preparation method of a microbial odor cleaner suitable for the environment, comprising the following steps:

[0121] Step one, according to the proportion, lactococcus lactis and bacillus velezensis are added into citric acid-disodium hydrogen phosphate buffer containing 0.5% mass fraction of sucrose, and activated at 35℃ for 2h;

[0122] Step two, according to the proportion, bromelain, lipase, rosemary acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture are added into the system obtained in step one, and homogenized at 30℃ for 5min;

[0123] Step three, according to the proportion, konjac glucomannan-chitosan gel and trehalose-glycerol complex are added, stirred, and packaged in the dark to obtain a microbial odor cleaner suitable for the environment.

[0124] Example 3

[0125] A microbial odor cleaner suitable for the environment, comprising the following raw material components by weight fraction:

[0126] Lactococcus lactis 25 parts;

[0127] Bacillus velezensis 25 parts;

[0128] Bromelain 10 parts;

[0129] Lipase 8 parts;

[0130] Rosemary acid nanoemulsion 20 parts;

[0131] Tea polyphenol 12 parts;

[0132] Citrus polyphenol-EDTA mixture 15 parts;

[0133] Konjac glucomannan-chitosan gel 18 parts;

[0134] Citric acid-disodium hydrogen phosphate buffer 30 parts;

[0135] Trehalose-glycerol complex 15 parts.

[0136] A preparation method of a microbial odor cleaner suitable for the environment, comprising the following steps:

[0137] Step one, according to the proportion, lactobacillus frumenti and bacillus velezensis are added to citric acid-sodium phosphate buffer solution containing 0.5% mass fraction of sucrose, and activated at 35℃ for 2h;

[0138] Step two, according to the proportion, bromelain, lipase, rosemary acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture are added to the system obtained in step one, and homogenized at 30℃ for 5min;

[0139] Step three, according to the proportion, konjac glucomannan-chitosan gel and trehalose-glycerol complex are added, stirred, and packed in the dark to obtain a microbial odor remover suitable for the environment.

[0140] Comparative example 1

[0141] A microbial odor remover suitable for the environment, comprising the following raw material components by weight fraction:

[0142] Lactobacillus frumenti 20 parts;

[0143] Bacillus velezensis 20 parts;

[0144] Bromelain 8 parts;

[0145] Lipase 5 parts;

[0146] Rosemary acid nanoemulsion 15 parts;

[0147] Tea polyphenol 8 parts;

[0148] Citrus polyphenol-EDTA mixture 12 parts;

[0149] Konjac glucomannan-chitosan gel 14 parts;

[0150] Citric acid-sodium phosphate buffer solution 25 parts;

[0151] Trehalose-glycerol complex 12 parts.

[0152] A preparation method of a microbial odor remover suitable for the environment, comprising the following steps:

[0153] Step one, according to the proportion, lactobacillus frumenti and bacillus velezensis are added to citric acid-sodium phosphate buffer solution containing 0.5% mass fraction of sucrose, and activated at 35℃ for 2h;

[0154] Step two, according to the proportion, bromelain, lipase, rosemary acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture are added to the system obtained in step one, and homogenized at 30℃ for 5min;

[0155] Step three, according to the proportion, konjac glucomannan-chitosan gel and trehalose-glycerol complex are added, stirred, and packed in the dark to obtain a microbial odor remover suitable for the environment.

[0156] The difference between the present comparative example and Example 1 is that the microencapsulated modified P. acidilactici is not added, but the unmodified original P. acidilactici is added.

[0157] Comparative Example 2

[0158] A microbial odor cleaner suitable for the environment includes the following raw material components by weight fraction:

[0159] P. acidilactici 20 parts;

[0160] Original B. velezensis 20 parts;

[0161] Papain 8 parts;

[0162] Lipase 5 parts;

[0163] Rosmarinic acid nanoemulsion 15 parts;

[0164] Tea polyphenol 8 parts;

[0165] Citrus polyphenol-EDTA mixture 12 parts;

[0166] Konjac glucomannan-chitosan gel 14 parts;

[0167] Citric acid-disodium hydrogen phosphate buffer 25 parts;

[0168] Trehalose-glycerol complex 12 parts.

[0169] A preparation method of a microbial odor cleaner suitable for the environment includes the following steps:

[0170] Step one, according to the ratio, P. acidilactici and original B. velezensis are added to citric acid-disodium hydrogen phosphate buffer containing 0.5% mass fraction of sucrose, and activated at 35°C for 2h;

[0171] Step two, according to the ratio, papain, lipase, rosmarinic acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture are added to the system obtained in step one, and homogenized at 30°C for 5min;

[0172] Step three, according to the ratio, konjac glucomannan-chitosan gel and trehalose-glycerol complex are added, stirred, and packaged in the dark to obtain a microbial odor cleaner suitable for the environment.

[0173] The difference between the present comparative example and Example 1 is that the B. velezensis with a polydopamine coating on the surface is not added, but the unmodified original B. velezensis is added.

[0174] Comparative Example 3

[0175] A microbial odor cleaner suitable for the environment, comprising the following raw material components by weight fraction:

[0176] Pediococcus acidilactici 20 parts;

[0177] Bacillus velezensis 20 parts;

[0178] Pronase 8 parts;

[0179] Lipase 5 parts;

[0180] Rosmarinic acid nanoemulsion 15 parts;

[0181] Tea polyphenol 8 parts;

[0182] Citrus polyphenol-EDTA mixture 12 parts;

[0183] Konjac glucomannan-chitosan gel 14 parts;

[0184] Citric acid-disodium hydrogen phosphate buffer 25 parts;

[0185] Trehalose-glycerol complex 12 parts.

[0186] A preparation method of a microbial odor cleaner suitable for the environment, comprising the following steps:

[0187] Step one, according to the proportion, Pediococcus acidilactici and Bacillus velezensis are added to citric acid-disodium hydrogen phosphate buffer containing 0.5% mass fraction of sucrose, and activated at 35°C for 2h;

[0188] Step two, according to the proportion, pronase, lipase, rosmarinic acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture are added to the system obtained in step one, and homogenized at 30°C for 5min;

[0189] Step three, according to the proportion, konjac glucomannan-chitosan gel and trehalose-glycerol complex are added, stirred, and packaged in the dark to obtain a microbial odor cleaner suitable for the environment.

[0190] The difference between the present comparative example and Example 1 is that ZIF-8 metal framework encapsulated bromelain is not added, but unmodified pronase is added.

[0191] Comparative Example 4

[0192] A microbial odor cleaner suitable for the environment, comprising the following raw material components by weight fraction:

[0193] Pediococcus acidilactici 20 parts;

[0194] Bacillus velezensis 20 parts;

[0195] Pronase 8 parts;

[0196] Protease 5 parts;

[0197] Rosmarinic acid nanoemulsion 15 parts;

[0198] Tea polyphenol 8 parts;

[0199] Citrus polyphenol-EDTA mixture 12 parts;

[0200] Konjac glucomannan-chitosan gel 14 parts;

[0201] Citric acid-disodium hydrogen phosphate buffer 25 parts;

[0202] Trehalose-glycerol complex 12 parts.

[0203] A preparation method of a microbial odor cleaner suitable for the environment, comprising the following steps:

[0204] Step one, according to the proportion, lactococcus lactis and bacillus velezensis are added into citric acid-disodium hydrogen phosphate buffer containing 0.5% mass fraction of sucrose, and activated at 35℃ for 2h;

[0205] Step two, according to the proportion, bromelain, pro-fat, rosmarinic acid nanoemulsion, tea polyphenol and citrus polyphenol-EDTA mixture are added into the system obtained in step one, and homogenized at 30℃ for 5min;

[0206] Step three, according to the proportion, konjac glucomannan-chitosan gel and trehalose-glycerol complex are added, stirred, and stored in dark place, to obtain a microbial odor cleaner suitable for the environment.

[0207] The difference between the present comparative example and example 1 is that the polyethylene glycol-phospholipid modified lipase is not added, but the unmodified pro-fat is added.

[0208] Test:

[0209] Fresh living organic garbage is selected, and is stored in a plastic bucket with a cover that can be sealed and can be evacuated. The volume of the garbage taken is about 1 / 2 to 2 / 3 of the volume of the barrel, and each barrel is labeled for easy inoculation. Then the barrel with garbage is stored in a closed room, the heater is turned on to ensure that the temperature in the room is maintained at 25±2℃, and fermentation is carried out for 11 days. The microbial odor cleaner prepared in examples 1-3 and comparative examples 1-4 is mixed with tap water at a mass ratio of 1:50 to prepare a preparation, which is sprayed for use. The ratio of the inoculum is 1g of preparation per 100g of garbage.

[0210] In the mixed gas of garbage producing foul odor, the content of ammonia and hydrogen sulfide is relatively high, so ammonia and hydrogen sulfide are used as detection indexes, and the specific detection method is as follows:

[0211] I. Ammonia determination: According to the sodium hypochlorite-salicylic acid spectrophotometric method (GB / T 14679-1993), the principle is that ammonia is absorbed by dilute sulfuric acid to form ammonium sulfate. In the presence of sodium nitroprusside, ammonium ions, salicylic acid and sodium hypochlorite react to form a blue compound, and the absorbance is measured at a wavelength of 697 nm using a spectrophotometer according to the color depth.

[0212] According to the measured absorbance, the ammonia content is calculated, and the removal rate of ammonia is calculated, and the calculation formula is as follows:

[0213] η N = [(C0-C1) / C0 x 100%

[0214] η N - ammonia removal rate; C0- ammonia content of the blank sample; C1- ammonia content after treatment;

[0215] The test results are shown in Table 1.

[0216] II. Hydrogen sulfide determination: According to the methylene blue spectrophotometric method (GB / T 11742-1989), the principle is that hydrogen sulfide in the air is absorbed by an alkaline cadmium hydroxide suspension to form cadmium sulfide precipitate. The addition of polyvinyl alcohol ammonium phosphate in the absorption solution can reduce the photolysis of cadmium sulfide. Then, in sulfuric acid solution, hydrogen sulfide reacts with p-aminodimethyl aniline solution and ferric chloride solution to form methylene blue. The color depth is colorimetrically quantified.

[0217] According to the measured absorbance, the hydrogen sulfide content is calculated, and the removal rate of hydrogen sulfide is calculated, and the calculation formula is as follows:

[0218] η S = [(C2-C3) / C2] x 100%

[0219] η S - hydrogen sulfide removal rate; C2- hydrogen sulfide content of the blank sample; C3- hydrogen sulfide content after treatment; The test results are shown in Table 1.

[0220] Table 1

[0221]

[0222] From Table 1, compared with Comparative Examples 1-4, the prepared formulations of Examples 1-3 have high degradation capacity for ammonia, hydrogen sulfide and other substances in odor.

[0223] Although embodiments of the present application have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and alterations can be made thereto without departing from the principles and spirit of the present application, the scope of which is defined by the appended claims and their equivalents.

[0224] In the description, references to "one embodiment," "an example," "certain examples" etc. mean that the particular feature, structure, material, or characteristic being described is included in at least one embodiment or example of the application. The appearances of an item in various examples or embodiments is not necessarily indicative of a frequency of occurrence in the various examples or embodiments. Moreover, descriptions of well-known structures, materials or methods can be omitted so as to not obscure the description of the subject application. In the description, the terms "couple" or "coupled" and "connect" or "connected," mean an indirect or direct electrical association or link.

[0225] The foregoing is merely illustrative of the principles of the application and various modifications can be made by persons skilled in the art without departing from the scope of the application as defined by the appended claims.

Claims

1. A microbial odor eliminator suitable for the environment, characterized by: The following raw materials are included in parts by weight: 15-25 parts of Pediococcus acidilactici; 15-25 parts of Bacillus Velez; 5-10 parts bromelain; 3-8 parts of lipase; 10-20 parts of rosmarinic acid nanoemulsion; 5-12 parts of tea polyphenols; 8-15 parts of citrus polyphenol-EDTA mixture; 10-18 parts of konjac glucomannan-chitosan gel; 20-30 parts of citric acid-disodium hydrogen phosphate buffer; 8-15 parts of trehalose-glycerol complex.

2. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The Pediococcus acidilactici needs to be microencapsulated with sodium alginate, and the specific operation is as follows: Pediococcus acidilactici was inoculated into MRS liquid medium with a pH of 6.2 and fermented at 37°C for 24 h until the bacterial concentration was ≥1×10 9 CFU / mL; the bacterial solution was mixed with a 2.5% sodium alginate solution, and a 2% CaCl2 solution was added dropwise through a microcapsule generator. After solidification for 15 minutes, the microcapsules were collected and freeze-dried for storage; The volume ratio of the bacterial solution to the sodium alginate solution is 1:3; The voltage of the microcapsule generator is 6-10 kV, and the diameter of the needle is 0.5 mm.

3. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The surface of the Bacillus Velez is coated with polydopamine, and the specific operation is as follows: Bacillus velez is cultured in a GYS medium containing 0.1% by mass of MnSO4 at 48-52°C with shaking for 36-48 hours to obtain a spore suspension; the spore suspension is mixed with a polydopamine solution having a concentration of 0.1-0.3 mg / mL, the pH is adjusted to 8.5, the reaction is carried out for 3-6 hours, and after centrifugation and washing, a Bacillus velez with a polydopamine adhesive coating on the surface is obtained; The volume ratio of the spore suspension to the polydopamine solution is 1:(3-5).

4. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The bromelain needs to be encapsulated in a ZIF-8 metal framework, and the specific operation is as follows: Dissolve Zn(NO3)2·6H2O in methanol solution in an ice-water bath at 2-6°C, adjust the pH to 6-8, add bromelain, add a methanol solution containing 2-methylimidazole, stir for 8-10 minutes, let stand for 2-5 minutes, collect the precipitate by centrifugation, wash, and dry to obtain ZIF-8-encapsulated bromelain; The methanol solution is prepared by methanol and pure water in a volume ratio of 3:7; The dosage ratio of the Zn(NO3)2·6H2O, methanol, bromelain, 2-methylimidazole and methanol solution is (14.5-15.3) g:500 mL: (1.8-2.2) g: (3-3.6) g:1000 mL.

5. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The lipase needs to be modified with polyethylene glycol-phospholipid, and the specific operation is as follows: The lipase was mixed with 1,2-distearoyl-phosphatidylethanolamine-polyethylene glycol in a phosphate buffer solution with a pH of 7.4, and the mixture was shaken at 35°C for 3 hours. The free phospholipids were removed by ultrafiltration, and the mixture was freeze-dried to obtain the polyethylene glycol-phospholipid-modified lipase. The molecular weight of the polyethylene glycol in the 1,2-distearoyl-phosphatidylethanolamine-polyethylene glycol is 2000; The dosage ratio of the lipase, 1,2-distearoyl-phosphatidylethanolamine-polyethylene glycol and phosphate buffer is 1 g: (18-22) g: (200-300) mL.

6. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The preparation steps of the rosemary nanoemulsion are as follows: Mix rosmarinic acid, Tween 80, and isopropyl myristate and heat to 60-70°C; slowly add 60-70°C pure water and homogenize at 10,000-12,000 rpm for 8-12 minutes; cool to room temperature and sterilize through a 0.22 μm filter membrane; The mass ratio of the rosmarinic acid, Tween 80, isopropyl myristate and pure water is 1:4:1:

54.

7. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The preparation steps of the citrus polyphenol-EDTA mixture are as follows: Dissolve citrus polyphenols and disodium ethylenediaminetetraacetic acid in ethanol solution, stir and react at 55-65°C for 6-8 hours, rotary evaporate at 40-50°C, and freeze-dry to obtain a citrus polyphenol-EDTA mixture; The total phenol content of the citrus polyphenols is greater than or equal to 60%; The mass ratio of the citrus polyphenols to disodium edetate is (1.6-2):1; The ethanol solution is prepared by mixing ethanol and pure water in a volume ratio of 1:1; The usage ratio of the citrus polyphenols to the ethanol solution is 1 g: (20-30) mL.

8. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The preparation steps of described konjac glucomannan-chitosan gel are as follows: Mixing konjac glucomannan and chitosan, adding acetic acid solution with a mass fraction of 1% acetic acid at 30-40° C., adjusting the pH to 5-5.5, and stirring for 15-30 minutes to obtain konjac glucomannan-chitosan gel; The deacetylation degree of the chitosan is ≥90%; The usage ratio of the konjac glucomannan, chitosan and acetic acid solution is 1g:(1-1.2)g:(30-50)mL.

9. The microbial odor eliminator suitable for use in an environment according to claim 1, characterized in that: The preparation steps of described konjac glucomannan-chitosan gel are as follows: Mixing konjac glucomannan and chitosan, adding acetic acid solution with a mass fraction of 1% acetic acid at 30-40° C., adjusting the pH to 5-5.5, and stirring for 15-30 minutes to obtain konjac glucomannan-chitosan gel; The usage ratio of the konjac glucomannan, chitosan and acetic acid solution is 1g:(1-1.2)g:(30-50)mL.

10. A method for preparing a microbial odor eliminator suitable for use in an environment as claimed in any one of claims 1 to 9, comprising the following steps: Step 1: adding Pediococcus acidilactici and Bacillus Velezii into a citric acid-disodium hydrogen phosphate buffer solution containing 0.5% by mass of sucrose according to the ratio, and standing at 30-40° C. for 2-4 hours for activation; Step 2: Add bromelain, lipase, rosmarinic acid nanoemulsion, tea polyphenols and citrus polyphenol-EDTA mixture according to the ratio to the system obtained in step 1, and homogenize at 28-32°C for 3-8 minutes; Step 3: Add konjac glucomannan-chitosan gel and trehalose-glycerol complex according to the ratio, stir, and package in a dark place to prepare a microbial odor eliminator suitable for the environment.

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