A bio-enzyme reinforcing liquid, a preparation method and application thereof

Abstract

The present application relates to a kind of biological enzyme strengthening liquid and its preparation method and application, by weight parts, include the following components: crude glycerol 10-40 parts, soybean meal 3-6 parts, corn flour 3-6 parts, yeast fermentation liquor 5-40 parts, inorganic composite carrier 1-5 parts, composite biological bacteria 0.5-2 parts, composite biological enzyme 0.1-1 part and water 10-50 parts;The biological enzyme strengthening liquid of the present application is by a large number of glyceric acid small molecules homogenization carbon source and nutrient factor, and the co-metabolic degradation effect of microorganism is strengthened, with corresponding long-chain alkane degrading bacteria and extracellular enzyme, the decomposition of substrate pollutants is strengthened, trace element and electron donor activate the activity of indigenous bacteria, quickly form the symbiotic, metabolic cooperation of specific flora system, multi-collaboration, and then enhance the system resistance to impact load capacity, the COD degradation effect of oily wastewater is remarkable.

Description

Technical Field

[0001] This invention relates to a bio-enzyme-enhanced liquid for wastewater treatment, its preparation method and application, and particularly to a bio-enzyme-enhanced liquid that is especially suitable for treating oily wastewater, its preparation method and application, belonging to the field of environmental protection technology. Background Technology

[0002] With the development of society and economy, industrial pollution has become one of the major sources of pollution in today's society. Among industrial pollution, oily wastewater from petroleum enterprises is the most serious. Oily wastewater contains large amounts of oil, heavy metals, long-chain aromatic hydrocarbons, and other toxic organic matter that is difficult to degrade. When discharged into the soil, it forms an oil film, making it difficult for air to penetrate, hindering the proliferation of soil microorganisms, destroying the soil's granular structure, and posing a great threat to crop growth. When petrochemical wastewater is discharged into water bodies, the floating oil forms an oil film that hinders atmospheric reoxygenation. The decomposition of emulsified and dissolved oils consumes dissolved oxygen in the water, causing anoxic conditions, increasing CO2 concentration, and decreasing pH levels, which poses a huge threat to aquatic life. In the past 50 years, more than 1,000 marine species have become extinct due to oil pollution, and marine life has decreased by 40%. Direct ingestion of petroleum distillates by humans can cause various poisoning symptoms, affecting organs such as the lungs, gastrointestinal tract, kidneys, central nervous system, and hematopoietic system.

[0003] Enzymes are a class of biological catalysts. Thousands of enzymes exist within living organisms, governing numerous catalytic processes such as metabolism, nutrient and energy conversion. Most reactions closely related to life processes are enzyme-catalyzed. Enzymes participate in various physiological and biochemical activities of organisms. They possess unique catalytic capabilities, each performing its specific function within the cell, without substitution for others. Examples include amylase digesting starch, protease digesting proteins, and synthetic enzymes promoting substance synthesis. Biological enzymes can improve the utilization rate of pollutants in wastewater by lowering activation energy, accelerating biological reactions, and promoting metabolism, providing the energy needed for bacterial growth.

[0004] Currently, the removal of organic matter from oily wastewater mainly relies on biological treatment as the core process. When biologically treating various organic substances in oily wastewater, microorganisms complete complex energy and synthetic metabolism to achieve pollutant degradation. However, oily wastewater has complex water quality, contains toxic substances such as heavy metals, and has a limited range of nutrients. The number of bacterial species that can degrade long-chain alkanes is insufficient, and naturally growing bacteria cannot synthesize all the enzymes necessary for growth and reproduction. The poor activity of the bacteria makes it difficult to achieve the ideal metabolic treatment effect, which manifests as activated sludge deflocculation, decreased effluent transparency, sludge bulking, and increased effluent COD. Therefore, a large amount of activated carbon needs to be added for adsorption treatment, which is costly. Summary of the Invention

[0005] This invention addresses problems encountered in oily wastewater treatment such as activated sludge deflocculation, decreased effluent transparency, sludge bulking, and increased effluent COD by providing a bio-enzyme-enhanced liquid, its preparation method, and its application.

[0006] A bio-enzyme enhancement solution, by weight, comprises the following components:

[0007] 10-40 parts crude glycerol, 3-6 parts soybean meal, 3-6 parts corn flour, 5-40 parts yeast fermentation broth, 1-5 parts inorganic composite carrier, 0.5-2 parts compound bacteria, 0.1-1 parts compound enzymes and 10-50 parts water;

[0008] The preparation method of the yeast fermentation broth is as follows: Wash and peel the potatoes, cut them into pieces, boil them until soft, then crush them. Add 0.1-1 wt% agar according to the weight of the potatoes, stir and mix well. After cooling, add 5-10 times the amount of water, then add 5-10 wt% pineapple juice and 0.05-0.5 wt% yeast, stir thoroughly, and ferment at 30-35℃ for 3-7 days. Then filter to obtain the yeast fermentation broth.

[0009] The inorganic composite carrier comprises the following components: activated carbon, vermiculite powder, tourmaline powder, and hydroxyl cellulose ether, wherein the mass ratio of activated carbon, vermiculite powder, tourmaline powder, and hydroxyl cellulose ether is 30:40:15:2.

[0010] The composite microbial organism consists of *Pseudomonas denitrificans*, *Bacillus cereus*, *Pseudomonas aeruginosa*, and *Acinetobacter jumbo*. The *Pseudomonas denitrificans* is deposited at the China General Microbiological Culture Collection Center (CGMCC) under accession number CGMCC No. 24446. The ratio of viable counts of *Pseudomonas denitrificans*, *Bacillus cereus*, *Pseudomonas aeruginosa*, and *Acinetobacter jumbo* is 1:(0.3-0.5):(0.6-0.8):(1.2-1.5), and the total viable count in the composite microbial organism is not less than 10. 8 CFU / g;

[0011] The complex bioenzyme is composed of β-mannanase, lipase and laccase, with a mass ratio of 1:(0.4-0.5):(0.6-0.7).

[0012] Preferably, the method for preparing the pineapple juice is as follows: cut the pineapple into 1-2cm pieces. 3 Crush the pineapple into small pieces, then soak it in water at a mass ratio of 1:(3-8) for 20-30 minutes, and filter to obtain pineapple juice.

[0013] The preparation method of the aforementioned bio-enzyme-enhanced solution includes the following steps:

[0014] 1) Wash and peel the potatoes, cut them into chunks, boil them until soft, then crush them. Add 0.1-1 wt% agar according to the weight of the potatoes, stir well, cool, add 5-10 times the amount of water, then add 5-10 wt% pineapple juice and 0.05-0.5 wt% yeast, stir thoroughly, and ferment at 30-35℃ for 3-7 days. Then filter to obtain the yeast fermentation liquid.

[0015] 2) According to the weight proportions, add the weighed crude glycerol, soybean meal, corn flour, inorganic composite carrier, composite bacteria, composite enzyme and water into the reaction vessel, and stir at a constant temperature of 30-60℃ for 36-72h; 3) Add the weighed yeast fermentation broth into the reaction vessel, and continue stirring at a constant temperature of 35-50℃ for 48-72h. Filter to remove impurities to obtain the enzyme-enhanced solution.

[0016] Preferably, the method for preparing the pineapple juice is as follows: cut the pineapple into 1-2cm pieces. 3 After crushing the pineapple into small pieces, soak it in water at a mass ratio of 1:(3-8) for 20-30 minutes, then filter to obtain pineapple juice.

[0017] Preferably, the stirring speed in steps 2) and 3) is 150-170 r / min.

[0018] The present invention also claims protection for the application of the above-mentioned bio-enzyme-enhanced solution in wastewater treatment.

[0019] Preferably, the wastewater treatment refers to the treatment of oily wastewater.

[0020] Preferably, the amount of bio-enzyme-enhanced liquid added to the oily wastewater is 20-100 ppm.

[0021] The working principle of the bio-enzyme enhancement solution of this invention is as follows:

[0022] 1) Yeast can produce various active extracellular enzymes and killing factors such as cellulase, alkaline protease, amylase, lipase, and phytase. Combined with special degrading bacteria and corresponding biological enzymes added during the production process, it degrades the recalcitrant components in wastewater, enhances the decomposition of matrix pollutants, and reacts with oily substances in wastewater to break down their chains, modify them, and precipitate them. It decomposes the recalcitrant large organic molecules in wastewater into small organic molecules. The small organic molecules are used and consumed by the native compound degrading bacteria as energy substances, thereby achieving the degradation of COD in wastewater.

[0023] 2) Yeast contains more than 20 kinds of amino acids and a variety of vitamins, with an extremely rich content of B vitamins. Its fermentation products are rich in nutrients and growth factors. Combined with the trace elements provided by the inorganic composite carrier and the enhancement effect of electron donors, it can provide the nutrients needed for the growth of native strains, enhance the activity of native strains, and promote their ability to degrade pollutants in wastewater.

[0024] The beneficial effects of the bio-enzyme-enhancing solution provided by this invention are as follows:

[0025] 1) By using a large amount of homogenized carbon sources of small molecules such as glyceric acid and nutrient factors to enhance the co-metabolic degradation of microorganisms, combined with special degrading bacteria and corresponding biological enzymes to enhance the degradation of recalcitrant components in wastewater, the activity of native bacteria is activated, and a microbial community system with specific symbiotic and metabolic cooperative relationships is quickly formed. The synergistic effect of multiple bacteria enhances the system's resistance to shock loads, and the COD degradation effect of oily wastewater is significant.

[0026] 2) The bio-enzyme-enhanced liquid of the present invention is suitable for large-scale industrial production and has a wide range of applications, especially for oily wastewater with rich organic matter content, such as oilfield drainage and oilfield mud filtrate. It has low manufacturing cost and is easy to use. Attached Figure Description

[0027] Figure 1 Analysis of microbial community structure at the phylum level (left: before addition, right: after addition)

[0028] Figure 2 Analysis of microbial community structure at the genus level (left: before addition, right: after addition).

[0029] Figure 3 The COD value is the value monitored online by the system terminal during the pilot-scale test.

[0030] Figure 4 This refers to the ammonia nitrogen value monitored online by the system terminal during the pilot-scale test. Detailed Implementation

[0031] The principles and features of the present invention are described below with reference to examples. The examples are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0032] The inorganic composite carriers used in Examples 1-4 were prepared by mixing activated carbon, vermiculite powder, tourmaline powder and hydroxycellulose ether in a mass ratio of 30:40:15:2 and passed through a 4000-mesh sieve.

[0033] The ratio of viable counts of *Pseudomonas denitrificans*, *Bacillus cereus*, *Pseudomonas aeruginosa*, and *Acinetobacter jumbo* in the compound microbial culture was 1:0.4:0.7:1.3, and the total viable count in the compound microbial culture was (20-30)×10⁻⁶. 8 CFU / g; Bacillus cereus, Pseudomonas aeruginosa, and Acinetobacter juncus can be purchased from any manufacturer or supplier on the market, while Pseudomonas denitrificans was purchased from the China General Microbiological Culture Collection Center, with accession number CGMCC No. 24446.

[0034] The complex bioenzyme is composed of β-mannanase, lipase and laccase, with a mass ratio of 1:(0.4-0.5):(0.6-0.7).

[0035] Example 1: Preparation of Bioenzyme Enhancement Solution

[0036] (1) Cultivation of yeast fermentation broth:

[0037] Wash and peel 200g of potatoes, then cut them into small pieces. Add 200g of water and boil for 30 minutes. Then, add the potatoes to a blender and blend at 1800rpm. Add 0.4g of agar and continue heating and stirring until well mixed. After cooling, add water to bring the volume to 1000ml. Add 10g of pineapple juice and 0.2g of yeast, and stir thoroughly. Incubate at 30℃ in a shaker at 160rpm for 7 days. The viable cell count in the fermentation broth should be no less than 10⁻⁶ after incubation. 8 CFU / g; (2) Preparation of enzyme hydrolysate:

[0038] Add 40g crude glycerol, 4g soybean meal, 4g corn flour, 2g inorganic composite carrier, 1g composite bacteria, 1g composite enzyme and 50g water to the reaction vessel, and react for 36h at a constant temperature of 60℃ and a stirring speed of 150r / min.

[0039] (3) Preparation of bio-enzyme-enhanced solution:

[0040] Add 40g of the yeast fermentation broth obtained in step (1) to the reaction vessel in step (2), and continue stirring and reacting at a constant temperature of 35℃ for 72h. Filter to remove impurities to obtain bio-enzyme enhanced solution 1. Example 2: Preparation of bio-enzyme enhanced solution

[0041] (1) Cultivation of yeast fermentation broth:

[0042] Wash and peel 200g of potatoes, then cut them into small pieces. Add 200g of water and boil for 30 minutes. Then, blend the potatoes in a blender at 1800rpm, add 0.2g of agar, and continue heating and stirring until well mixed. After cooling, add water to bring the volume to 1000ml. Add 20g of pineapple juice and 0.4g of yeast, and stir thoroughly. Incubate at 35℃ in a shaker at 160r / min for 5 days. The viable cell count in the fermentation broth should be no less than 10. 8 CFU / g;

[0043] (2) Preparation of enzyme hydrolysate:

[0044] Add 30g crude glycerol, 3g soybean meal, 3g corn flour, 1g inorganic composite carrier, 1g composite bacteria, 0.5g composite enzyme, and 30g water to a reaction vessel and react for 36 hours at a constant temperature of 50℃ and a stirring speed of 150r / min.

[0045] (3) Preparation of bio-enzyme-enhanced solution:

[0046] Add 25g of the yeast fermentation broth obtained in step (1) to the reaction vessel in step (2), and continue stirring and reacting at a constant temperature of 40℃ for 48h. Filter to remove impurities to obtain bio-enzyme enhanced solution 2. Example 3: Preparation of bio-enzyme enhanced solution

[0047] (1) Cultivation of yeast fermentation broth:

[0048] Wash and peel 200g of potatoes, then cut them into small pieces. Add 200g of water and boil for 30 minutes. Then, add the potatoes to a blender and blend at 1800rpm. Add 1.0g of agar and continue heating and stirring until well mixed. After cooling, add water to bring the volume to 1000ml. Add 10g of pineapple juice and 1g of yeast, and stir thoroughly. Incubate at 30℃ in a shaker at 160rpm for 7 days. The viable cell count after incubation should be no less than 10. 8 CFU / g; (2) Preparation of enzyme hydrolysate:

[0049] Add 20g crude glycerol, 6g soybean meal, 6g corn flour, 5g inorganic composite carrier, 0.5g composite bacteria, 0.4g composite enzyme, and 20g water to a reaction vessel and react for 72h at a constant temperature of 30℃ and a stirring speed of 150r / min.

[0050] (3) Preparation of bio-enzyme-enhanced solution:

[0051] Add 15g of the yeast fermentation broth obtained in step (1) to the reaction vessel in step (2), and continue stirring and reacting at a constant temperature of 50℃ for 60h. Filter to remove impurities to obtain the bioenzyme-enhanced solution 3.

[0052] Example 4: Preparation of bio-enzyme enhancement solution:

[0053] (1) Cultivation of yeast fermentation broth:

[0054] Wash and peel 200g of potatoes, then cut them into small pieces. Add 200g of water and boil for 30 minutes. Then, add the potatoes to a blender and blend at 1800rpm. Add 0.6g of agar and continue heating and stirring until well mixed. After cooling, add water to bring the volume to 1000ml. Add 20g of pineapple juice and 0.8g of yeast, and stir thoroughly. Incubate at 35℃ in a shaker at 160rpm for 3 days. The viable cell count in the fermentation broth should be no less than 10⁻⁶ after incubation. 8 CFU / g; (2) Preparation of enzyme hydrolysate:

[0055] Add 10g of crude glycerol, 6g of soybean meal, 6g of corn flour, 2g of inorganic composite carrier, 2g of composite bacteria, 0.1g of composite enzyme, and 10g of water to a reaction vessel, and react for 36 hours at a constant temperature of 55℃ and a stirring speed of 150r / min.

[0056] (3) Preparation of bio-enzyme-enhanced solution:

[0057] Add 5g of the yeast fermentation broth obtained in step (1) to the reaction vessel in step (2), and continue stirring and reacting at a constant temperature of 50℃ for 60h. Filter to remove impurities to obtain the bioenzyme-enhanced solution 4.

[0058] Example 5: Application Effect of Oilfield Wastewater Discharge

[0059] The bio-enzyme-enhanced solutions provided in Examples 1-4 of this invention are used for the treatment of oilfield wastewater. During wastewater treatment, the stock solution is diluted to the appropriate concentration based on the properties of the bio-enzyme-enhanced solution and the degree of water pollution before being added. For severely polluted wastewater, the stock solution can be added directly. The dosage is mainly determined by the degree of water pollution, the addition point, and the results of small-scale tests; typically, the dosage is 20-100 ppm.

[0060] The testing methods for wastewater treatment performance are as follows:

[0061] Effluent from the integrated biological treatment tank of the oilfield wastewater treatment plant, with an initial COD of 300 mg / L, was added to SBR reaction systems 1-5. To simulate the aerobic treatment system, 30% sludge from the integrated biological treatment tank was added to each system. The bio-enzyme enhancement solution provided in Examples 1-4 was added to SBR reaction systems 1-4 at a ratio of 100 mg of bio-enzyme enhancement solution per liter of wastewater. SBR reaction system 5 was not added to the bio-enzyme enhancement solution and served as a blank control. Under conditions of 25℃ and dissolved oxygen of 2 mg / L, two parallel experiments were conducted for each test subject. The COD degradation rate in the oilfield wastewater was measured at 48h, 72h, and 96h after addition, and the average value was taken. The test results are shown in Table 1.

[0062] Table 1: Results of COD Degradation Rate Measurement

[0063] Test object 48h COD degradation rate % 72h COD degradation rate % 96h COD degradation rate % Example 1 73 87 94 Example 2 72 85 93 Example 3 75 86 91 Example 4 74 85 90 control group 65 75 84

[0064] Based on the data analysis in Table 1, the following conclusions can be drawn:

[0065] The bio-enzyme-enhanced liquid prepared in Examples 1-4 of this invention can be used to treat oily wastewater, effectively improving the degradation efficiency of pollutants and increasing the COD degradation rate of wastewater from 84% to over 90%.

[0066] Example 6: Production Dosing Effect

[0067] On September 3, 2023, the Shucai Wastewater Treatment Brigade introduced the bio-enzyme enhancement solution of this invention into the heavy oil wastewater treatment system of the Shuguang Wastewater Treatment Plant for a production addition test, aiming to enhance the comprehensive microbial capacity (quantity, types, activity, etc.) of the existing PACT process. The experiment ended on October 31, 2023.

[0068] The wastewater to be treated first enters a two-stage air flotation system, where the floating oil and fine suspended solids are coagulated into flocs by the action of coagulants. After the flocs are separated and removed, the wastewater enters the equalization tank and cooling tower to adjust the water quality and quantity. Then it is lifted into the integrated biological system (PACT process) for carbonization and nitrification reactions. During the pilot test, biological enzyme enhancement solution 1 is added at a dosage of 40 ppm to the first-stage distribution mixing tank at the front end of the integrated biological system.

[0069] Microbial community structure analysis of the activated sludge system before and after pilot-scale testing, detection of the abundance and species changes of dominant bacteria, and high-throughput analysis of community structure components, such as... Figure 1 As shown, the width represents the relative abundance of different genera; only species with higher abundance are displayed, while the remaining species are grouped together and represented in a different format. Figure 1 , Figure 2 It is evident that after adding bio-enzyme enhancement solution to the system, the proportion of dominant bacteria at the phylum and genus levels decreased, microbial diversity increased, other strains were added and were able to colonize the system, and gradually developed into new dominant bacteria.

[0070] The bio-enzyme enhancement solution used in this pilot test acts on microorganisms, and the enhancement effect on microorganisms is directly reflected in the change in chemical oxygen demand (COD) removal rate. During the pilot test, the average COD value monitored at the system terminal / online was 20.47 mg / L. Figure 3 As shown, compared with the average value of 28.49 mg / L in the early stage of the trial (January-August 2023), the decrease was 28.2%.

[0071] The ammonia nitrogen level in the wastewater effluent from the wastewater treatment system fluctuated significantly. Although the average value of 0.43 mg / L in the early stage of the experiment (January-August 2023) was far below the required standard for compliance, the average ammonia nitrogen level during the experiment was 0.07 mg / L. Figure 4 As shown, the reduction rate reached 83.7% compared with the early stage of the experiment, which indirectly reflects that the removal effect of ammonia nitrogen becomes more and more obvious with the increase of microbial species.

[0072] The conclusions drawn from the pilot-scale data analysis are as follows:

[0073] The bio-enzyme-enhanced liquid prepared in Example 1 of this invention can enhance the comprehensive microbial capacity of the heavy oil wastewater treatment system. The ammonia nitrogen index of the terminal effluent of the heavy oil wastewater treatment system decreased by ≥50% compared with the average value in the early stage of the experiment, and the actual ammonia nitrogen reduction in the pilot test was 83.7%.

[0074] The bio-enzyme-enhanced liquid prepared by this invention enhances the co-metabolic degradation of microorganisms by providing homogenized carbon sources and nutrient factors. Combined with corresponding long-chain alkane-degrading bacteria and extracellular enzymes, it enhances the decomposition of matrix pollutants. Trace elements and electron donors activate the activity of local bacterial species, rapidly forming a microbial community system with a specific symbiotic relationship. The synergistic effect of multiple bacteria results in significant pollutant degradation.

[0075] 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 bio-enzyme fortification solution, characterized in that, By weight, it contains the following components: 10-40 parts crude glycerol, 3-6 parts soybean meal, 3-6 parts corn flour, 5-40 parts yeast fermentation broth, 1-5 parts inorganic composite carrier, 0.5-2 parts compound bacteria, 0.1-1 parts compound enzymes and 10-50 parts water. The preparation method of the yeast fermentation broth is as follows: Wash and peel the potatoes, cut them into pieces, boil them until soft, then crush them. Add 0.1-1 wt% agar according to the weight of the potatoes, stir and mix well. After cooling, add 5-10 times the amount of water, then add 5-10 wt% pineapple juice and 0.05-0.5 wt% yeast, stir thoroughly, and ferment at 30-35℃ for 3-7 days. Then filter to obtain the yeast fermentation broth. The inorganic composite carrier comprises the following components: activated carbon, vermiculite powder, tourmaline powder, and hydroxyl cellulose ether, wherein the mass ratio of activated carbon, vermiculite powder, tourmaline powder, and hydroxyl cellulose ether is 30:40:15:

2. The composite microbial organism consists of *Pseudomonas denitrificans*, *Bacillus cereus*, *Pseudomonas aeruginosa*, and *Acinetobacter jumbo*. The *Pseudomonas denitrificans* is deposited at the China General Microbiological Culture Collection Center (CGMCC) under accession number CGMCC No. 24446. The ratio of viable counts of *Pseudomonas denitrificans*, *Bacillus cereus*, *Pseudomonas aeruginosa*, and *Acinetobacter jumbo* is 1:(0.3-0.5):(0.6-0.8):(1.2-1.5), and the total viable count in the composite microbial organism is not less than 10. 8 CFU / g; The complex bioenzyme is composed of β-mannanase, lipase and laccase, with a mass ratio of 1:(0.4-0.5):(0.6-0.7).

2. The bio-enzyme-enhancing solution according to claim 1, characterized in that, The method for preparing the pineapple juice is as follows: cut the pineapple into 1-2cm pieces. 3 Crush the pineapple into small pieces, then soak it in water at a mass ratio of 1:(3-8) for 20-30 minutes, and filter to obtain pineapple juice.

3. A method for preparing a bio-enzyme-enhanced solution, characterized in that, Includes the following steps: 1) Wash and peel the potatoes, cut them into chunks, boil them until soft, then crush them. Add 0.1-1 wt% agar according to the weight of the potatoes, stir well, cool, add 5-10 times the amount of water, then add 5-10 wt% pineapple juice and 0.05-0.5 wt% yeast, stir thoroughly, and ferment at 30-35℃ for 3-7 days. Then filter to obtain the yeast fermentation liquid. 2) By weight, add 10-40 parts of crude glycerol, 3-6 parts of soybean meal, 3-6 parts of corn flour, 1-5 parts of inorganic composite carrier, 0.5-2 parts of composite bacteria, 0.1-1 parts of composite enzyme and 10-50 parts of water to the reaction vessel, and stir at a constant temperature of 30-60℃ for 36-72 hours. The inorganic composite carrier comprises the following components: activated carbon, vermiculite powder, tourmaline powder, and hydroxyl cellulose ether, wherein the mass ratio of activated carbon, vermiculite powder, tourmaline powder, and hydroxyl cellulose ether is 30:40:15:

2. The composite microbial organism consists of *Pseudomonas denitrificans*, *Bacillus cereus*, *Pseudomonas aeruginosa*, and *Acinetobacter jumbo*. The *Pseudomonas denitrificans* is deposited at the China General Microbiological Culture Collection Center (CGMCC) under accession number CGMCC No. 24446. The ratio of viable counts of *Pseudomonas denitrificans*, *Bacillus cereus*, *Pseudomonas aeruginosa*, and *Acinetobacter jumbo* is 1:(0.3-0.5):(0.6-0.8):(1.2-1.5), and the total viable count in the composite microbial organism is not less than 10. 8 CFU / g; The composite bioenzyme is composed of β-mannanase, lipase and laccase, with a mass ratio of 1:(0.4-0.5):(0.6-0.7). 3) Add 5-40 parts of the weighed yeast fermentation broth to the reaction vessel, and continue stirring and reacting at a constant temperature of 35-50℃ for 48-72 hours. Filter to remove impurities to obtain the bio-enzyme enhanced solution.

4. The preparation method according to claim 3, characterized in that, The method for preparing the pineapple juice is as follows: cut the pineapple into 1-2cm pieces. 3 Crush the pineapple into small pieces, then soak it in water at a mass ratio of 1:(3-8) for 20-30 minutes, and filter to obtain pineapple juice.

5. The preparation method according to claim 3 or 4, characterized in that, In steps 2) and 3), the stirring speed is 150-170 r / min.

6. The application of the bio-enzyme-enhanced liquid according to claim 1 in wastewater treatment.

7. The application of the bio-enzyme-enhanced liquid according to claim 2 in wastewater treatment.

8. The application according to claim 6 or 7, characterized in that, The wastewater in question refers to oily wastewater.

9. The application according to claim 8, characterized in that, The amount of bio-enzyme-enhanced solution added to the oily wastewater is 20-100 ppm.

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