Biological oil and odor removing agent for oil separation tank and preparation method thereof

Abstract

The application provides a biological oil removal and odor removal agent for an oil separation tank and a preparation method thereof, and belongs to the technical field of oil removal and odor removal agents. The biological oil removal and odor removal agent comprises the following raw materials in parts by weight: slow-release bacterial balls 50-70 parts, fermentation products 15-25 parts, and a composite surfactant 5-10 parts. The composite surfactant comprises a first surfactant and a second surfactant, and the mass ratio of the first surfactant to the second surfactant is 3-5:10. The biological oil removal and odor removal agent for the oil separation tank can better separate and emulsify oil stains and reduce oil stain residues. Meanwhile, lipase enzymatically hydrolyzes the oil stains to generate free fatty acids, relevant bacteria act on the fatty acids to promote further oxidation reaction of the fatty acids, and carbon dioxide and water are generated. The biological oil removal and odor removal agent also has a good bacteriostatic effect. The active bacteria in the application are embedded in microspheres, which can effectively prevent the inactivation of the active bacteria. After being added into water, the microsphere shells slowly break, the active bacteria are slowly released, and the active bacteria and the microspheres jointly play a role in better oil removal and odor removal, and therefore, the biological oil removal and odor removal agent has a wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of oil and odor removal agents, and particularly relates to a biological oil and odor removal agent for an oil separation tank and a preparation method thereof. BACKGROUND

[0002] The rapid development of the catering industry has generated a large amount of wastewater containing oil and fat. After the oil and fat enters the drainage pipeline, it is cooled and solidified and adheres to the inner wall of the pipeline, causing the pipeline to narrow or even be blocked. If a large amount of oil and fat enters the water body, it will form an oil film to block oxygen exchange, causing aquatic organisms to die of hypoxia and destroying the ecology. The accumulated oil and fat provides a breeding environment for mice, cockroaches, mosquitoes and flies, and bacteria breed, increasing the risk of disease transmission. The oil separation tank for catering is a key equipment for kitchen wastewater treatment. Its core advantage is to separate oil and fat, suspended solids and food residues from wastewater through physical separation principle, effectively reducing the risk of pipeline blockage, reducing the load of sewage treatment plants, and avoiding the damage of direct discharge of oil and fat to the water ecology. However, there are still significant pain points in its actual application:

[0003] 1. Odor emission: The long-stored oil and fat is prone to corruption and fermentation under high temperature, breeding anaerobic bacteria and releasing hydrogen sulfide and other foul-smelling gases, affecting the kitchen environment and surrounding air quality;

[0004] 2. Fluctuation of oil separation efficiency: The traditional oil separation tank has poor separation effect on small oil droplets, and the oil-water separation efficiency is low when the flow is overloaded or the oil is seriously emulsified;

[0005] 3. Complex cleaning and maintenance: The accumulated oil needs to be manually removed regularly (usually 1-2 times per week), and the operation process is dirty and easy to cause secondary pollution. Some equipment with unreasonable structure also has the problem of oil accumulation in dead corners which is difficult to completely remove.

[0006] Due to these pain points of the oil separation tank, more effective methods are needed to reduce the oil and environmental odor entering the drainage pipeline. SUMMARY

[0007] The present application aims to provide a biological oil and odor removal agent for an oil separation tank and a preparation method thereof, which can better separate and emulsify oil and dirt, reduce oil and dirt residues, and at the same time, lipase can enzymatically hydrolyze oil and dirt to generate free fatty acids. The related bacteria act on the fatty acids to promote further oxidation reaction to generate carbon dioxide and water. The biological oil and odor removal agent also has good antibacterial effect. The active bacteria in the present application are embedded in microspheres, which can effectively prevent the inactivation of the active bacteria. After being added to water, the microsphere shell slowly breaks and releases the active bacteria, which work together to better remove oil and odor, and have a broad application prospect.

[0008] The technical scheme of the present application is as follows:

[0009] The application provides a bio-oil and odor removing agent for oil separation tanks, which comprises the following raw materials in parts by weight: slow-release bacterial balls 50-70 parts, fermentation products 15-25 parts, and a composite surfactant 5-10 parts, wherein the composite surfactant comprises a first surfactant and a second surfactant in a mass ratio of 3-5:10; the first surfactant is C10-16 alkyl derivatized benzene sulfonic acid or C12-15 fatty alcohol polyoxyethylene ether; and the second surfactant has a structural formula as shown in formula I.

[0010]

[0011] Formula I

[0012] wherein m=3-6.

[0013] As a further improvement of the application, the preparation method of the slow-release bacterial balls and the fermentation products is as follows:

[0014] S1. Preparation of a culture medium: glyceryl trioleate, glycerol, proteose peptone, yeast extract, inorganic salt, alanine, methionine and glutathione are added into deionized water, sterilized, and a culture medium is prepared;

[0015] S2. Activation of bacterial strains: Pseudomonas fluorescens CICC 22031, Pseudomonas putida CICC 20576, Bacillus subtilis CICC 10732, Bacillus licheniformis CICC 10291, Bacillus thuringiensis CICC 20556 and Bacillus velezensis CICC 24560 are inoculated into LB culture medium respectively, activated and cultured, and bacterial seed solutions are prepared;

[0016] S3. Fermentation culture: the bacterial seed solutions are inoculated into the culture medium, and fermentation culture is carried out; filtration, washing and freeze-drying are carried out to obtain enhanced bacterial bodies; the filtrate is layered, the water layer is collected, and freeze-drying is carried out to prepare fermentation products;

[0017] S4. Preparation of slow-release bacterial balls: gelatin and sodium alginate are dissolved in water, the enhanced bacterial bodies are added, stirring and mixing are carried out, an emulsifier is added, fish oil is added dropwise, emulsification is carried out, calcium chloride solution is added dropwise, normal temperature solidification is carried out, centrifugation is carried out, washing is carried out, and freeze-drying is carried out to prepare slow-release bacterial balls.

[0018] As a further improvement of the present invention, in step S1, the mass ratio of trioleic acid glyceride, glycerol, peptone, yeast extract, inorganic salts, alanine, methionine, and glutathione is 15-20:5-10:1-2:1-3:1-2:0.5-1:0.2-0.5:0.1-0.3, and the inorganic salts include magnesium sulfate, calcium chloride, and ferric chloride, with a mass ratio of 2-4:3-5:1-3; in step S2, the activation culture temperature is 25-30℃, the speed is 100-200 r / min, and the time is 18-30 h, and the bacterial count of the inoculum is 10. 8 -10 9 cfu / mL.

[0019] As a further improvement of the present invention, the *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyssus* CICC mentioned in step S3 are also included. The inoculation amounts of the 24560 bacterial spore liquid were 0.5-1 v / v%, 0.5-1 v / v%, 0.5-1 v / v%, 0.5-1 v / v%, 1-1.5 v / v%, and 1-1.5 v / v%. The fermentation culture temperature was 25-30℃, 100-200 r / min, and fermentation culture time was 36-48 h. In step S4, the mass ratio of gelatin, sodium alginate, cell enhancer, and emulsifier was 7-10:8-12:5-10:0.1-0.3. The emulsifier was selected from at least one of Tween-20, Tween-40, Tween-60, Tween-80, and Tween-85.

[0020] As a further improvement of the present invention, the method for synthesizing the second surfactant is as follows:

[0021] T1. Dehydroabsic acid, catalyst, and thionyl chloride were mixed and heated to prepare intermediate 1, with the following structure: ;

[0022] T2. Intermediate 1 was reacted with 3-dimethylaminopropylamine and a base to obtain intermediate 2, with the following structure: ;

[0023] T3. Perfluorooctanoic acid and benzoyl chloride are mixed and reacted to prepare perfluorooctanoyl chloride, with the following structure: CF3(CF2)6COCl;

[0024] T4. Intermediate 3 was prepared by reacting perfluorooctanoyl chloride, 3-dimethylaminopropylamine, and a base, with the following structure: ;

[0025] T5. Intermediate 2, intermediate 3 and dibromoalkane are mixed and reacted to obtain the product.

[0026] As a further improvement of the present invention, the catalyst in step T1 is 4-dimethylaminopyridine, the molar ratio of dehydroabietic acid to thionyl chloride is 1:1.2-1.5, the amount of catalyst added is 15-20% of the molar amount of dehydroabietic acid, the heating reaction temperature is 70-75°C, and the time is 2-4 h; in step T2, the molar ratio of intermediate 1 and 3-dimethylaminopropylamine to the base is 1:1-1.2:2-4, the reaction time is 1-3 h, and the base is triethylamine, NaOH, or KOH.

[0027] As a further improvement of the present invention, the molar ratio of perfluorooctanoic acid and benzoyl chloride in step T3 is 1:1-1.2; the molar ratio of perfluorooctanoic acid chloride, 3-dimethylaminopropylamine, and base in step T4 is 1:1-1.2:2-4, the reaction time is 1-3 hours, and the base is triethylamine, NaOH, or KOH.

[0028] As a further improvement of the present invention, the molar ratio of intermediate 2, intermediate 3 and dibromoalkane in step T5 is 1:1-1.1:1, the reaction temperature is 75-85℃, and the reaction time is 44-52h. The dibromoalkane is selected from at least one of 1,3-dibromopropane, 1,4-dibromobutane, 1,5-dibromopentane, and 1,6-dibromohexane.

[0029] This invention further protects a method for preparing the above-mentioned biological oil removal and deodorizing agent for grease traps, comprising the following steps:

[0030] (1) The first surfactant and the second surfactant are mixed evenly to obtain a composite surfactant;

[0031] (2) Mix the slow-release bacterial balls, fermentation products and compound surfactant evenly to prepare a biological oil removal and deodorizing agent for oil separators.

[0032] The present invention further protects a method of using the above-mentioned biological oil-removing and deodorizing agent for grease traps, wherein the biological oil-removing and deodorizing agent for grease traps is mixed with water to prepare a 5-10 wt% suspension, which is then dripped into the grease trap for oil removal and deodorization treatment.

[0033] The present invention has the following beneficial effects:

[0034] The culture medium of this invention contains multiple components, including glycerol and trioleic acid glycerides as carbon sources. It also includes glucose-free components to maintain cell growth and avoid the "glucose effect" that inhibits lipase synthesis. Glycerol also acts as an osmotic regulator, helping cells maintain intracellular osmotic balance under adverse conditions such as high osmotic pressure, preventing excessive cell dehydration and damage, thereby improving the stress resistance of the fermenting bacteria. A complex nitrogen source (including peptone and yeast extract) is used to control the carbon-to-nitrogen ratio, promoting the shift of metabolic flux to lipase synthesis during enzyme production. Peptone is rich in nitrogen and trace elements, which helps cell growth and reproduction, improving cell survival rate and stress resistance. Yeast extract is rich in protein, amino acids, vitamins, and minerals, providing comprehensive nutrition for the fermenting bacteria, promoting cell growth and metabolism, and enhancing cell stress resistance. Mg is added to the inorganic salts of this invention. 2+ Ca 2+ Stabilize enzyme structure, Fe 3+ It promotes electron transport chain activity and enhances enzyme activity. The addition of alanine and methionine as amino acid precursors enhances ribosome synthesis efficiency; glutathione, an important antioxidant, can scavenge reactive oxygen free radicals within the bacteria, reduce the damage of oxidative stress to the bacteria, maintain cell stability and function, thereby improving the stress resistance of the fermenting bacteria and increasing yeast's tolerance to reactive oxygen species; potassium dihydrogen phosphate provides phosphorus and potassium to the fermenting bacteria, participates in the metabolic processes of the bacteria, regulates intracellular acid-base balance and osmotic pressure, and enhances the stress resistance of the bacteria.

[0035] Biological additives provide a large number of microbial strains, which decompose the oils in wastewater by producing a series of metabolic enzymes. In particular, lipases can effectively catalyze the conversion of triglycerides into glycerol and free fatty acids. Since lipases are interfacial activated enzymes, they can only function at the oil (lipid)-water interface. The enzyme is only activated when it comes into contact with the oil-water interface.

[0036] The bacteria of this invention comprise *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyss* CICC 24560. The *Bacillus* produce enzymes that break down fats into glycerol and fatty acids; the *Pseudomonas* further decompose the fatty acids, oxidize unsaturated fatty acids, and decompose them into carbon dioxide and water under sufficient oxygen supply. Using only *Bacillus* poses a risk, such as excessive free fatty acids. In such cases, high calcium levels in the surrounding environment can easily lead to the formation of calcium fatty acid deposits in drainage pipes, gradually causing accumulation. Therefore, the synergistic effect of *Bacillus* and *Pseudomonas* can significantly enhance the oil removal capacity.

[0037] To further enhance interfacial interaction, this invention employs a combination of biological additives and surfactants. The surfactants disperse large oil droplets into micron-sized emulsion droplets, increasing the contact area for lipases and accelerating the enzymatic hydrolysis reaction. The biological additives also eliminate the production of malodorous gases such as hydrogen sulfide from oil spoilage, thus reducing unpleasant odors.

[0038] This invention prepares a second surfactant with strong aggregation ability, good water solubility, good foaming properties, and good calcium soap dispersibility. The preparation method is simple, the synthesis conditions are mild, and the yield is high. Containing a rosin structure, it exhibits antibacterial or bacteriostatic effects against fungi or other molds in decaying wood, further reducing odor and purifying the environment. A hydrophobic flexible chain is introduced into the rigid framework of rosin through an amide group, and a fluorocarbon chain is introduced at the other end, exhibiting good hydrophobic and oleophobic properties. Its oleophobicity is much stronger than that of hydrocarbon chains, making the fluorocarbon-containing surfactant more approachable to and interacting with oil stains, thereby better dissolving and dispersing oil stains and significantly improving the degreasing effect. The fluorocarbon-containing surfactant can penetrate deeper into the oil stains, separating and emulsifying the oil stains from the cleaning surface, reducing oil residue. Simultaneously, the fluorocarbon surfactant can reduce the interfacial tension between oil and water, weakening the interaction between oil and water, making it easier to remove oil stains. It forms a low surface energy molecular film in aqueous solution, reducing the surface tension of the solution, allowing the surfactant to better wet the oily surface, increasing the contact area with the oil stains, and creating favorable conditions for subsequent degreasing processes. The first and second surfactants of this invention have a synergistic effect. Although the surfactants have a certain antibacterial effect and may have a negative impact on the active bacteria of this invention, during storage, the active agent does not come into contact with the surfactant in the microspheres, so the impact is minimal. After being formulated into a solution, the active bacteria can initially exert a highly efficient biological degreasing and deodorizing effect.

[0039] The biological oil and odor removal agent for grease traps prepared by this invention can effectively separate and emulsify oil, reducing oil residue. At the same time, lipase enzymatically hydrolyzes the oil to generate free fatty acids. Related bacteria act on the fatty acids, promoting further oxidation to generate carbon dioxide and water, and also have a good antibacterial effect. The active bacteria of this invention are encapsulated in microspheres, which can effectively prevent their inactivation. After being added to water, the microsphere shells slowly rupture, releasing the active bacteria in a slow-release manner, which work together to better remove oil and odor, and has broad application prospects. Detailed Implementation

[0040] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0041] Preparation Example 1: Preparation of Slow-Release Bacterial Spheres and Fermentation Products

[0042] The method is as follows:

[0043] S1. Preparation of culture medium: 15g trioleic acid glyceride, 5g glycerol, 1g peptone, 1g yeast extract, 1g inorganic salt, 0.5g alanine, 0.2g methionine, and 0.1g glutathione were added to 250mL of deionized water and sterilized to obtain the culture medium.

[0044] The inorganic salts include magnesium sulfate, calcium chloride, and ferric chloride in a mass ratio of 2:3:1.

[0045] S2. Activation of bacterial strains: *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyss* CICC 24560 were inoculated into LB medium and activated at 25°C and 100 rpm for 18 h, yielding a bacterial count of 10... 8 -10 9 CFU / mL bacterial seed solution;

[0046] S3. Fermentation Culture: Seed cultures of *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belye* CICC 24560 were inoculated into the culture medium at inoculation rates of 0.5 v / v%, 0.5 v / v%, 0.5 v / v%, 0.5 v / v%, 1 v / v%, and 1 v / v%, respectively. The culture was carried out at 25°C and 100 r / min for 36 h. After filtration, washing, and freeze-drying, enhanced bacterial cells were obtained. The filtrate was allowed to stand and separate into layers. The aqueous layer was collected, freeze-dried, and the fermentation product was obtained.

[0047] S4. Preparation of slow-release bacterial balls: Dissolve 7g of gelatin and 8g of sodium alginate in 250mL of water, add 5g of enhanced bacterial cells, stir and mix evenly, add 0.1g of Tween-20, dropwise add to 500mL of fish oil, emulsify at 8000r / min for 15min, dropwise add 10mL of 5wt% calcium chloride solution, solidify at room temperature for 30min, centrifuge, wash, freeze dry to obtain slow-release bacterial balls.

[0048] Preparation Example 2: Preparation of Slow-Release Bacterial Spheres and Fermentation Products

[0049] The method is as follows:

[0050] S1. Preparation of culture medium: 20g trioleic acid glyceride, 10g glycerol, 2g peptone, 3g yeast extract, 2g inorganic salt, 1g alanine, 0.5g methionine, and 0.3g glutathione were added to 250mL of deionized water and sterilized to obtain the culture medium.

[0051] The inorganic salts include magnesium sulfate, calcium chloride, and ferric chloride in a mass ratio of 4:5:3.

[0052] S2. Activation of bacterial strains: *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyss* CICC 24560 were inoculated into LB medium and activated at 30°C and 200 rpm for 30 h, yielding a bacterial count of 10... 8 -10 9 CFU / mL bacterial seed solution;

[0053] S3. Fermentation Culture: Seed cultures of *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belye* CICC 24560 were inoculated into the culture medium at inoculation amounts of 1 v / v%, 1 v / v%, 1 v / v%, 1 v / v%, 1.5 v / v%, and 1.5 v / v%, respectively. Fermentation was carried out at 30℃ and 200 r / min for 48 h. After filtration, washing, and freeze-drying, enhanced bacterial cells were obtained. The filtrate was allowed to stand and separate into layers; the aqueous layer was collected and freeze-dried to obtain the fermentation product.

[0054] S4. Preparation of slow-release bacterial balls: Dissolve 10g of gelatin and 12g of sodium alginate in 250mL of water, add 10g of enhanced bacterial cells, stir and mix evenly, add 0.3g of Tween-40, dropwise add to 500mL of fish oil, emulsify at 8000r / min for 15min, dropwise add 10mL of 5wt% calcium chloride solution, solidify at room temperature for 30min, centrifuge, wash, freeze dry to obtain slow-release bacterial balls.

[0055] Preparation Example 3: Preparation of Slow-Release Bacterial Spheres and Fermentation Products

[0056] The method is as follows:

[0057] S1. Preparation of culture medium: 17g trioleic acid glyceride, 7g glycerol, 1.5g peptone, 2g yeast extract, 1.5g inorganic salt, 0.7g alanine, 0.35g methionine, and 0.2g glutathione were added to 250mL of deionized water and sterilized to obtain the culture medium.

[0058] The inorganic salts include magnesium sulfate, calcium chloride, and ferric chloride in a mass ratio of 3:4:2.

[0059] S2. Activation of bacterial strains: *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyss* CICC 24560 were inoculated into LB medium and activated at 27°C and 150 rpm for 22 h, yielding a bacterial count of 10... 8 -10 9 CFU / mL bacterial seed solution;

[0060] S3. Fermentation Culture: Seed cultures of *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyssiensis* CICC 24560 were inoculated into the culture medium at inoculation rates of 0.7 v / v%, 0.7 v / v%, 0.7 v / v%, 0.7 v / v%, 1.2 v / v%, and 1.2 v / v%, respectively. Fermentation was carried out at 27°C and 150 rpm for 42 h. The cultures were then filtered, washed, and freeze-dried to obtain enhanced bacterial cells. The filtrate was allowed to separate into layers, the aqueous layer was collected, and freeze-dried to obtain the fermentation product.

[0061] S4. Preparation of slow-release bacterial balls: Dissolve 8.5g of gelatin and 10g of sodium alginate in 250mL of water, add 7g of enhanced bacterial cells, stir and mix evenly, add 0.2g of Tween-85, add dropwise to 500mL of fish oil, emulsify at 8000r / min for 15min, add 10mL of 5wt% calcium chloride solution, solidify at room temperature for 30min, centrifuge, wash, freeze dry to obtain slow-release bacterial balls.

[0062] Comparative Preparation Example 1

[0063] The difference from Preparation Example 3 is that Pseudomonas spp. was not inoculated in step S3.

[0064] Specifically as follows:

[0065] S3. Fermentation culture: Bacillus subtilis CICC 10732, Bacillus licheniformis CICC 10291, Bacillus thuringiensis CICC 20556, and Bacillus belyssus CICC 24560 were fermented.

[0066] The inoculum seed solution was inoculated into the culture medium at inoculation amounts of 1.4% v, 1.4% v, 1.2% v, and 1.2% v, respectively. The culture was carried out at 27°C and 150 rpm for 42 hours. The culture was then filtered, washed, and freeze-dried to obtain enhanced cells. The filtrate was allowed to stand and separate into layers. The aqueous layer was collected, freeze-dried, and the fermentation product was obtained.

[0067] Comparative Preparation Example 2

[0068] The difference from Preparation Example 3 is that Bacillus spp. was not inoculated in step S3.

[0069] Specifically as follows:

[0070] S3. Fermentation culture: Seed culture of *Pseudomonas fluorescens* CICC 22031 and *Pseudomonas putida* CICC 20576 was inoculated into the culture medium at inoculation rates of 2.6 v / v%, 2.6 v / v%, 27℃, 150 r / min, and fermented for 42 h. The culture was then filtered, washed, and freeze-dried to obtain enhanced bacterial cells. The filtrate was allowed to stand and separate into layers. The aqueous layer was collected, freeze-dried, and the fermentation product was obtained.

[0071] Comparative preparation example 3

[0072] The difference compared to preparation example 3 is that step S4 was not performed.

[0073] S1. Preparation of culture medium: 17g trioleic acid glyceride, 7g glycerol, 1.5g peptone, 2g yeast extract, 1.5g inorganic salt, 0.7g alanine, 0.35g methionine, and 0.2g glutathione were added to 250mL of deionized water and sterilized to obtain the culture medium.

[0074] The inorganic salts include magnesium sulfate, calcium chloride, and ferric chloride in a mass ratio of 3:4:2.

[0075] S2. Activation of bacterial strains: *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyss* CICC 24560 were inoculated into LB medium and activated at 27°C and 150 rpm for 22 h, yielding a bacterial count of 10... 8 -10 9 CFU / mL bacterial seed solution;

[0076] S3. Fermentation culture: Seed cultures of *Pseudomonas fluorescens* CICC 22031, *Pseudomonas putida* CICC 20576, *Bacillus subtilis* CICC 10732, *Bacillus licheniformis* CICC 10291, *Bacillus thuringiensis* CICC 20556, and *Bacillus belyssiensis* CICC 24560 were inoculated into the culture medium at inoculation rates of 0.7 v / v%, 0.7 v / v%, 0.7 v / v%, 0.7 v / v%, 1.2 v / v%, and 1.2 v / v%, respectively. Fermentation was carried out at 27°C and 150 r / min for 42 h. The cultures were then filtered, washed, and freeze-dried to obtain enhanced bacterial cells. The filtrate was allowed to stand and separate into layers. The aqueous layer was collected, freeze-dried, and the fermentation product was obtained.

[0077] Preparation Example 4: Synthesis of the Second Surfactant

[0078] The method is as follows:

[0079] T1. 0.1 mol dehydroabietic acid and 0.02 mol 4-dimethylaminopyridine were mixed and added to a container. Thionyl chloride was added dropwise, and the mixture was heated to 75°C and stirred for 3 hours. After the reaction was completed, excess thionyl chloride was removed under reduced pressure to obtain intermediate 1. ESI-MS calculated value: C 20 H 28 ClO(M+H) + 319.18, measured value: 319.2, yield: 89%.

[0080] MRI results: 1 H NMR (300MHz, CDCl3) δ6.8-6.95 (m, 3H), 3.15 (m, 1H), 2.85 (t, 2H), 1.84-1.90 ( m, 4H), 1.75 (t, 1H), 1.62 (t, 2H), 1.42 (m, 2H), 1.37 (s, 3H), 1.23-1.25 (m, 9H);

[0081] Synthesis route:

[0082]

[0083] T2. 0.11 mol of 3-dimethylaminopropylamine and 0.3 mol of triethylamine were mixed and added to a container. 100 mL of dichloromethane solution containing 0.1 mol of intermediate 1 was added, and the mixture was stirred for 2 h. The reaction product was washed three times with weakly alkaline water, extracted three times with dichloromethane, and the dichloromethane and triethylamine were removed under reduced pressure to obtain intermediate 2. ESI-MS calculated value: C 25 H 41 N₂O(M+H) + 385.31, measured value: 385.3, yield: 80%.

[0084] MRI results: 1 H NMR (300MHz, CDCl3) δ8.0 (br, 1H) 6.85-6.99 (m, 3H), 3.19-3.32 (m, 3H), 2.85 (t, 2H), 2.25-2.37 (m, 8 H), 1.84-1.90 (m, 4H), 1.77 (t, 1H), 1.60-1.69 (t, 4H), 1.44 (m, 2H), 1.37 (s, 3H), 1.23-1.25 (m, 9H);

[0085] Synthesis route:

[0086]

[0087] T3. Mix 0.1 mol of perfluorooctanoic acid and 0.11 mol of benzoyl chloride, heat to 80°C, stir and react for 1 h, collect the fraction at 125-135°C, which is perfluorooctanoic acid chloride, with a melting point of 74-75°C and a yield of 96%;

[0088] Synthesis route:

[0089]

[0090] T4. 0.11 mol of 3-dimethylaminopropylamine and 0.3 mol of triethylamine were mixed and added to a container. 100 mL of a chloroform solution of perfluorooctanoyl chloride was added, and the mixture was stirred for 2 h. The reaction product was washed three times with weakly alkaline water, extracted three times with chloroform, and dichloromethane and triethylamine were removed under reduced pressure to obtain intermediate 3. ESI-MS calculated value: C 13 H 14 F 15 N₂O(M+H) + 499.08, measured value: 499.1, yield: 83%.

[0091] MRI results: 1 H NMR (300MHz, CDCl3) δ8.0 (br, 1H), 3.25 (t, 2H), 2.39 (t, 2H), 2.14 (s, 6H), 1.58 (m, 2H).

[0092] Synthesis route:

[0093]

[0094] T5. Add 0.1 mol of intermediate 2, 0.11 mol of intermediate 3, and 0.1 mol of 1,4-dibromobutane to a container, add 200 mL of ethanol, heat to 80 °C, stir for 50 h, remove the solvent under reduced pressure, recrystallize three times from ethyl acetate / ethanol, wash, and dry to obtain the product. ESI-MS calculated value: C 42 H 62 Br2F 15 N4O2(M+H) + 1097.29, measured value: 1097.3, yield: 75%.

[0095] MRI results: 1 H NMR (300MHz, CDCl3) δ8.0 (br, 2H), 6.8-6.92 (m, 3H), 3.15-3.35 (m, 25H), 2.95 (m, 2H), 1.92 (m, 4H), 1.5-1.8 (m, 11H), 1.12-1.27 (m, 14H).

[0096] Synthesis route:

[0097]

[0098] Example 1

[0099] This embodiment provides a method for preparing a biological oil removal and deodorizing agent for grease traps, comprising the following steps:

[0100] (1) C10-16 alkyl-derived benzenesulfonic acid and the second surfactant prepared in Preparation Example 4 were mixed evenly, wherein the mass ratio of C10-16 alkyl-derived benzenesulfonic acid and the second surfactant was 3:10, to obtain a composite surfactant;

[0101] (2) Mix 50g of the slow-release bacterial pellets prepared in Preparation Example 1, 15g of the fermentation product prepared in Preparation Example 1, and 5g of the composite surfactant evenly to prepare a biological oil-removing and deodorizing agent for grease traps. Prepare a 5wt% suspension by adding the biological oil-removing and deodorizing agent to the mixed solution. The mixed solution has the following formulation: 1wt‰ magnesium sulfate + 1wt‰ calcium chloride + 1wt‰ ferric chloride + 1wt‰ phosphate buffer + 1wt‰ magnesium sulfate + 1wt‰ calcium chloride + 1wt‰ ferric chloride + 1wt‰ phosphate buffer. Add the solution dropwise to the oil, and allow it to react for 12 hours to remove oil and odor. The addition rate is 100mL of suspension per 10mL of oil. Dry the remaining liquid and analyze the decomposition state of the oil components using Fourier transform infrared spectroscopy. The C=O stretching vibration of the ester bond (RCOOR') in intact triglycerides is at 1740 cm⁻¹. -1 A strong absorption peak is observed at this point, and the C=O vibration peak of free fatty acids shifts to lower frequencies to 1700-1720 cm⁻¹.-1 The products after the reaction were measured at 1711 cm⁻¹. -1 There is a peak nearby, and it is 1746cm high. -1 The decrease in peak area corresponding to the ester bond in fats and oils indicates that some fats and oils have decomposed into free fatty acids; the carboxylic acids (-COOH) and glycerol (-OH) produced by decomposition will be present at 3300 cm⁻¹. -1 A broad peak forms nearby, while the product after the reaction reaches 3457 cm⁻¹. -1 The peak intensity at 3471 cm⁻¹ is greater than that of intact oil. -1 The strength at that location.

[0102] Example 2

[0103] This embodiment provides a method for preparing a biological oil removal and deodorizing agent for grease traps, comprising the following steps:

[0104] (1) C10-16 alkyl-derived benzenesulfonic acid and the second surfactant prepared in Preparation Example 4 were mixed evenly, wherein the mass ratio of C10-16 alkyl-derived benzenesulfonic acid and the second surfactant was 5:10, to obtain a composite surfactant;

[0105] (2) Mix 70g of the slow-release bacterial balls prepared in Preparation Example 2, 25g of the fermentation product prepared in Preparation Example 2 and 10g of the composite surfactant evenly to prepare a biological oil removal and deodorizing agent for oil separator.

[0106] Example 3

[0107] This embodiment provides a method for preparing a biological oil removal and deodorizing agent for grease traps, comprising the following steps:

[0108] (1) C12-15 fatty alcohol polyoxyethylene ether and the second surfactant prepared in Preparation Example 4 were mixed evenly, and the mass ratio of C12-15 fatty alcohol polyoxyethylene ether to the second surfactant was 4:10, to obtain a composite surfactant.

[0109] (2) Mix 60g of the slow-release bacterial balls prepared in Preparation Example 3, 20g of the fermentation product prepared in Preparation Example 3 and 7g of the composite surfactant evenly to prepare a biological oil removal and deodorizing agent for oil separator.

[0110] Comparative Example 1

[0111] The difference from Example 3 is that the slow-release bacterial balls and fermentation products were both prepared from Comparative Preparation Example 1.

[0112] Comparative Example 2

[0113] The difference from Example 3 is that the slow-release bacterial balls and fermentation products were both prepared from Comparative Preparation Example 2.

[0114] Comparative Example 3

[0115] Compared with Example 3, the difference is that the fermentation products were all prepared by Comparative Preparation Example 3, and the slow-release bacterial balls were replaced by the enhanced bacterial cells prepared by Comparative Preparation Example 3 of equal mass.

[0116] Comparative Example 4

[0117] The difference from Example 3 is that the composite surfactant is replaced by a single C12-15 fatty alcohol polyoxyethylene ether.

[0118] Comparative Example 5

[0119] The difference from Example 3 is that the composite surfactant was replaced by a single second surfactant prepared in Preparation Example 4.

[0120] Comparative Example 6

[0121] The difference compared to Example 3 is that no compound surfactant was added.

[0122] Test Example 2: Determination of Surface Tension and Critical Micelle Concentration

[0123] Using deionized water as a solvent, the composite surfactants prepared in Examples 1-3 of this invention were formulated into solutions of different concentrations. The surface tension of the aqueous solutions was measured at room temperature using an integrated interfacial parameter measuring instrument. The obtained data were plotted as γ-lgC curves, and the inflection point of the curve was the critical micelle concentration (cmc) of the composition. The surface tension at the critical micelle concentration was also measured. A 50 mg / L solution was prepared, and the oil-water interfacial tension was measured at 65°C using a rotating drop interfacial tensiometer. The results are shown in Table 1.

[0124] Table 1

[0125]

[0126] As can be seen from the table above, the composite surfactant obtained in step (1) of Examples 1-3 of the present invention has good surface properties.

[0127] Test Example 2

[0128] The grease traps prepared in Examples 1-3 or Comparative Examples 1-3 were mixed with a biological oil-removing and deodorizing agent to prepare a 5wt% suspension. The mixed solution had the following formulation: 1wt‰ magnesium sulfate + 1wt‰ calcium chloride + 1wt‰ ferric chloride + 1wt‰ phosphate buffer + 1wt‰ magnesium sulfate + 1wt‰ calcium chloride + 1wt‰ ferric chloride + 1wt‰ phosphate buffer. This solution was added dropwise to the grease at a rate of 100mL of suspension per 10mL of grease, and the reaction was carried out for 12 hours to remove oil and odor. The remaining substances were measured, and the results are shown in Table 2.

[0129] Table 2

[0130]

[0131] As can be seen from the table above, the biological oil removal and deodorizing agents for oil separators prepared in Examples 1-3 of this invention can effectively degrade oils.

[0132] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A biological oil removal and deodorizing agent for grease traps, characterized in that, The composition comprises the following raw materials by weight: slow-release bacteria balls 50-70 parts, fermentation product 15-25 parts, and composite surfactant 5-10 parts, wherein the composite surfactant comprises a first surfactant and a second surfactant, and the mass ratio of the first surfactant to the second surfactant is 3-5:10; the first surfactant is C10-16 alkyl derivatized benzene sulfonic acid or C12-15 fatty alcohol polyoxyethylene ether, and the structural formula of the second surfactant is shown in formula I: Formula I wherein m = 3-6.

2. The bio-oil and odor removing agent for an oil separation tank according to claim 1, characterized by, The preparation method of the slow-release bacteria balls and the fermentation product is as follows: S1. Preparation of culture medium: glyceryl trioleate, glycerol, proteose peptone, yeast extract, inorganic salt, alanine, methionine, and glutathione are added to deionized water, sterilized, and a culture medium is prepared; S2. Activation of bacterial strains: Pseudomonas fluorescens CICC 22031, Pseudomonas putida CICC 20576, Bacillus subtilis CICC 10732, Bacillus licheniformis CICC 10291, Bacillus thuringiensis CICC 20556, and Bacillus velezensis CICC 24560 are inoculated into LB culture medium respectively, activated and cultured, and bacterial seed liquid is prepared; S3. Fermentation culture: the bacterial seed liquid is inoculated into the culture medium, fermented and cultured, filtered, washed, and freeze-dried to obtain enhanced bacteria; the filtrate is allowed to stand and stratify, the water layer is collected, and freeze-drying is performed to prepare a fermentation product; S4. Preparation of slow-release bacteria balls: gelatin and sodium alginate are dissolved in water, the enhanced bacteria are added, stirred and mixed uniformly, an emulsifier is added, and fish oil is added dropwise, emulsified, calcium chloride solution is added dropwise, and solidified at room temperature, centrifuged, washed, and freeze-dried to prepare slow-release bacteria balls.

3. The bio-oil and odor removing agent for an oil separation tank according to claim 2, characterized by In step S1, the mass ratio of glyceryl trioleate, glycerol, proteose peptone, yeast extract, inorganic salt, alanine, methionine, and glutathione is 15-20:5-10:1-2:1-3:1-2:0.5-1:0.2-0.5:0.1-0.3, and the inorganic salt comprises magnesium sulfate, calcium chloride, and ferric chloride, and the mass ratio is 2-4:3-5:1-3; in step S2, the temperature of the activated culture is 25-30℃, the rotation speed is 100-200 r / min, and the time is 18-30 h, and the bacterial content of the bacterial seed liquid is 108-109 cfu / mL.

4. The bio-oil and odor removing agent for an oil separation tank according to claim 2, characterized by The inoculation amount of the Pseudomonas fluorescens CICC 22031, Pseudomonas putida CICC 20576, Bacillus subtilis CICC 10732, Bacillus licheniformis CICC 10291, Bacillus thuringiensis CICC 20556 and Bacillus velezensis CICC 24560 seed liquid in step S3 is 0.5-1 v / v%, 0.5-1 v / v%, 0.5-1 v / v%, 0.5-1 v / v%, 1-1.5 v / v% and 1-1.5 v / v% respectively, the fermentation temperature is 25-30℃, the fermentation culture is 100-200 r / min, and the fermentation culture is 36-48 h; the mass ratio of the gelatin, sodium alginate, enhanced bacterial body and emulsifier in step S4 is 7-10:8-12:5-10:0.1-0.3, and the emulsifier is at least one selected from Tween-20, Tween-40, Tween-60, Tween-80 and Tween-85.

5. The bio-oil and odor removing agent for an oil separation tank according to claim 1, characterized by The synthesis method of the second surfactant is as follows: T1. Dehydroabietic acid, catalyst and thionyl chloride are mixed and heated to react to produce intermediate 1, the structure of which is as follows: ; T2. Mixing intermediate 1 and 3-dimethylaminopropylamine, a base, to produce intermediate 2, having the structure: ; T3. Full-fluorooctanoic acid and benzoyl chloride are mixed and reacted to prepare perfluorooctanoyl chloride, which has the following structure: CF3(CF2)6COCl; T4. The intermediate 3 is prepared by mixing perfluorooctanoyl chloride and 3- dimethylaminopropylamine, base, as shown below: ; T5. Intermediate 2, intermediate 3 and dibromoalkane are mixed and reacted to prepare the product.

6. The bio-oil and odor removing agent for an oil separation tank according to claim 5, characterized by The catalyst in step T1 is 4-dimethylaminopyridine, the molar ratio of the dehydroabietic acid and thionyl chloride is 1:1.2-1.5, the addition amount of the catalyst is 15-20% of the amount of substance of the dehydroabietic acid, the heating reaction temperature is 70-75℃, and the heating reaction time is 2-4 h; the molar ratio of the intermediate 1, 3-dimethylaminopropylamine and base in step T2 is 1:1-1.2:2-4, the reaction time is 1-3 h, and the base is triethylamine, NaOH or KOH.

7. The bio-oil and odor removing agent for an oil separation tank according to claim 5, characterized by The molar ratio of the full-fluorooctanoic acid and benzoyl chloride in step T3 is 1:1-1.2; the molar ratio of the perfluorooctanoyl chloride, 3-dimethylaminopropylamine and base in step T4 is 1:1-1.2:2-4, the reaction time is 1-3 h, and the base is triethylamine, NaOH or KOH.

8. The bio-oil and odor removing agent for an oil separation tank according to claim 5, characterized by, The molar ratio of the intermediate 2, intermediate 3 and dibromoalkane in step T5 is 1:1-1.1:1, the reaction temperature is 75-85℃, the reaction time is 44-52 h, and the dibromoalkane is at least one selected from 1,3-dibromopropane, 1,4-dibromobutane, 1,5-dibromopentane and 1,6-dibromohexane.

9. A method for producing a bio-oil and odor removing agent for an oil separator according to any one of claims 1 to 8, characterized by, The method comprises the following steps: (1) mixing the first surfactant and the second surfactant uniformly to prepare a composite surfactant; (2) mixing the slow-release bacterial ball, the fermentation product and the composite surfactant uniformly to prepare a biological oil and odor removal agent for an oil separation tank.

10. A method for using the bio-oil and odor removing agent for an oil separation tank according to any one of claims 1 to 8, characterized by, The biological oil and odor removal agent for an oil separation tank is prepared into a suspension of 5-10 wt% by adding water, and then is added dropwise into the oil separation tank for oil and odor removal treatment.

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