High-efficiency degradation of refractory organic matters in soy sauce wastewater and deep denitrification process and strain

Abstract

This invention provides a process and strain for removing recalcitrant organic matter and performing deep denitrification treatment on wastewater from Maotai liquor production. The specific process includes introducing wastewater into a regulating tank for water quality analysis; introducing the wastewater from the regulating tank into an anaerobic reactor; and introducing the effluent from the anaerobic reactor into an aerobic reactor, adding *Moraxella rosenbergii* (seawater). Rossellomorea aquimaris The process involves ZC2402-MT oxidation; the aerobic reaction effluent is then fed into an ozone reactor to convert the remaining recalcitrant organic nitrogen into ammonia nitrogen; the ozone effluent is then fed into a BAF (Biological Acid Fluidization) unit to remove remaining COD and perform nitrification; the BAF effluent is then fed into an autotrophic denitrification unit. This invention cultivates highly efficient degrading bacteria for the recalcitrant organic matter portion of liquor wastewater, while simultaneously adjusting the process to ensure that the effluent COD and total nitrogen meet standards, and to reduce costs, providing a more scientific, rational, and effective process for the treatment of liquor wastewater.

Description

Technical Field

[0001] This invention relates to the field of wastewater treatment technology, specifically to a highly efficient process and strain for degrading recalcitrant organic matter in liquor wastewater and for deep denitrification. Background Technology

[0002] Baijiu, a national industry and traditional biological industry with Chinese characteristics, has a history of thousands of years and is an outstanding cultural heritage of my country. In recent years, while promoting the increase in baijiu production and value, it has also caused certain environmental damage. Baijiu on the market is mainly divided into sauce-aroma baijiu and strong-aroma baijiu. Among them, sauce-aroma baijiu generates wastewater in all stages of production except crushing. The wastewater mainly consists of yellow water from fermentation on flat land, yellow water from the bottom of cellars, water from the bottom of distillation pots, and water from washing the ground. Its main pollutants include COD, BOD, NH3-N, SS, and color. Studies have shown that the effluent from sauce-aroma baijiu wastewater after treatment contains various complex organic compounds and proteins and long-chain hydrocarbons produced by biological reactions, which are difficult to biodegrade and cannot be removed by traditional biological processes alone. With the country's increasing efforts in ecological and environmental protection, some baijiu enterprises in environmentally sensitive areas are required to comply with the Class IV surface water discharge standard (the direct emission limit for COD is 30 mg / L, and the emission limit for total nitrogen is 1.5 mg / L).

[0003] Current technologies for treating brewing wastewater primarily rely on biochemical methods, supplemented by physical and chemical methods. The treatment process includes pretreatment, secondary treatment, and advanced treatment, with advanced treatment methods including adsorption, membrane filtration, advanced oxidation, and coagulation sedimentation. However, these combined processes suffer from long flow rates, large footprints, unstable effluent quality, and, most importantly, inability to meet current stringent effluent standards. Therefore, it is necessary to optimize traditional brewing wastewater treatment processes to make them more scientific, rational, and effective, while also meeting low-cost requirements. Summary of the Invention

[0004] To address the ecological and environmental requirements of certain liquor enterprises in environmentally sensitive areas, which necessitate compliance with Class IV surface water discharge standards (with direct COD emission limits of 30 mg / L and total nitrogen emission limits of 1.5 mg / L), this invention provides a process for removing recalcitrant organic matter and performing deep denitrification treatment on liquor wastewater, ensuring that the effluent meets the standards.

[0005] The primary objective of this invention is to provide a highly efficient process for degrading recalcitrant organic matter in liquor wastewater and for deep denitrification.

[0006] The second objective of this invention is to provide a strain of bacteria and its agent that are highly efficient at degrading recalcitrant organic matter in liquor wastewater.

[0007] To achieve the primary objective, the technical solution adopted by this invention is as follows:

[0008] A highly efficient process for degrading recalcitrant organic matter and deeply removing nitrogen from liquor wastewater specifically includes the following steps:

[0009] S1. Introduce the wastewater into the equalization tank, analyze and measure the water quality, and adjust the influent pH to 3.5-6;

[0010] S2. The wastewater from the equalization tank in step S1 is introduced into the anaerobic reactor for anaerobic reaction, with a retention time of 0.5-1 day.

[0011] S3. The effluent from the anaerobic reaction in step S2 is introduced into an aerobic reactor containing ordinary activated sludge, and microorganisms are added for degradation treatment. The microorganisms are *Roseolario Moraxella salina* (seawater). Rossellomorea aquimaris ZC2402-MT was deposited on May 10, 2024, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 30575;

[0012] S4. After filtration, the aerobic reaction effluent from step S3 enters the ozone reactor to convert the remaining recalcitrant organic nitrogen into ammonia nitrogen. In this process, an ozone catalyst, such as hydrogen peroxide, can be added.

[0013] S5. The ozone effluent from step S4 is fed into the BAF device to remove the remaining COD and carry out nitrification.

[0014] S6. The BAF effluent from step S5 is fed into an autotrophic denitrification device to achieve the goal of ultimate denitrification.

[0015] Preferably, in step S3, trace elements Fe, Cu, Mo, Zn, Co, and Mn are added to the ordinary activated sludge.

[0016] Preferably, the mass ratio of the trace elements is: Fe 0~1mg / L, Cu 0~1mg / L, Mo 0~1mg / L, Zn 0~1mg / L, Co 0~1mg / L, Mn 0~1mg / L.

[0017] Preferably, the mass ratio of the amount of *Roseolario Moraxella salina* ZC2402-MT added to the mass ratio of the inoculum of ordinary activated sludge is 0.1-10%.

[0018] Preferably, the aerobic reactor delivers wastewater from the bottom, and the temperature inside the aerobic reactor is 33~37℃, with the pH controlled at 7.2-7.8.

[0019] Preferably, the aerobic reaction device is supplemented with nutrients in the following proportions: NaHCO3 0~1%, MgSO4 0~1%, CaCl2 0~1%, yeast extract 0~0.1%, and the pH is maintained between 7.2 and 7.8.

[0020] Preferably, the BAF device described in step S5 uses lightweight packing material to reduce packing caking.

[0021] To achieve the second objective, the present invention adopts the following technical solution:

[0022] A strain of highly efficient biodegrading bacteria for liquor wastewater, characterized by its classification and naming as *Marine Roselliamora* (Marine Roselliamora). Rossellomorea aquimaris ZC2402-MT was deposited on May 10, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 30575.

[0023] The morphological and physiological-biochemical characteristics of the strain of this invention are as follows:

[0024] Physiological characteristics: colonies are round or oval, with irregular edges, opaque, moist and dull.

[0025] The bacterial cells are rod-shaped, heterotrophic aerobic, and Gram-positive.

[0026] Beneficial effects of this invention:

[0027] This invention cultivates highly efficient degrading bacteria for the recalcitrant organic matter in liquor wastewater. Simultaneously, the process is adjusted by incorporating ozone and BAF (biochemical oxygen demand) processes, reducing land occupation and solid waste generation. A subsequent autotrophic nitrification denitrification step further reduces carbon source input while achieving maximal denitrification, ensuring that effluent COD and total nitrogen meet standards, and lowering costs. This provides a more scientific, rational, and effective process for treating liquor wastewater. Attached Figure Description

[0028] Figure 1 A schematic diagram of the process of this invention;

[0029] The components include: 1. Anaerobic reactor; 2. Aerobic reactor; 3. Highly efficient degrading bacteria; 4. Ozone advanced oxidation device; 5. Ozone catalyst; 6. BAF device; 7. Lightweight packing material; and 8. Autotrophic denitrification device.

[0030] The biological material described in this invention is classified and named as *Marine Rosellariae* (or *Moraxella marineis*). Rossellomorea aquimarisZC2402-MT was deposited on May 10, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 30575.

[0031] To address the shortcomings of existing processes, this paper improves the traditional treatment process for Maotai liquor wastewater by providing a new technology for removing recalcitrant organic matter and achieving deep denitrification. This involves cultivating highly efficient degrading bacteria for the recalcitrant components and adjusting the process by incorporating ozone and BAF (Bipolar Alternating Current) technology. A catalyst is added to the advanced ozone oxidation stage to improve ozone efficiency. Specific packing materials are selected in the BAF unit to reduce caking, increase efficiency, and simultaneously reduce land area and solid waste generation. A subsequent autotrophic nitrification process ensures that the total nitrogen in the effluent meets standards, thereby reducing costs. Detailed Implementation

[0032] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. The technical solutions in the embodiments of the present invention will be clearly and completely described. 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 of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0033] The following examples illustrate the present invention further, but do not limit the scope of the invention.

[0034] Example 1: Isolation and purification of *Roseolariomoraxella salina* strain ZC2402-MT.

[0035] S1 activation: Pick a single colony of the highly efficient degrading bacteria of soy sauce liquor wastewater from a solid plate and transfer it to a culture medium containing soy sauce liquor wastewater (5 mL system). Shake at 100-120 rpm and incubate at 30-35℃ for 24-48 h until the logarithmic phase.

[0036] S2 Transfer: The activated bacterial solution for degrading soy sauce liquor wastewater in the logarithmic phase from Process 1 is transferred to a 500mL seed tank at an inoculation rate of 2-5% for cultivation. The temperature of the seed tank is maintained at 30-35℃, the rotation speed is maintained at 100-120 rpm, and the dissolved oxygen (DO) is controlled at 4-6 mg / L for 24-48 hours. This allows the bacteria to tolerate higher concentrations and significantly improves the complete degradation rate of soy sauce liquor wastewater.

[0037] S3 Expansion Culture: The degradation bacterial broth cultured in the seed tank of Process 2 is transferred to a 10L fermenter for expansion culture at an inoculation rate of 2%-5%. The composition of the fermenter culture medium is the same as that of the seed tank, and the physicochemical parameters are as follows: temperature 30-35℃, rotation speed 100-120rpm, dissolved oxygen 4-6mg / L, and fermentation time 48-72h. After fermentation, the effective viable bacteria count in the tank broth can reach more than 109 CFU / ml. After the fermentation broth is discharged from the tank, it is packaged in plastic buckets to obtain the highly efficient degradation bacterial agent for soy sauce liquor wastewater.

[0038] The physiological characteristics of this strain are as follows: colonies are round or oval with irregular edges, opaque, moist, and dull. The bacterial cells are rod-shaped, heterotrophic aerobic, and Gram-positive.

[0039] S1. Introduce the wastewater into the equalization tank, analyze and measure the water quality, and adjust the influent pH to 3.5-6;

[0040] S2. The wastewater from the equalization tank in step S1 is introduced into the anaerobic reactor for anaerobic reaction, with a retention time of 0.5-1 day.

[0041] S3. The effluent from the anaerobic reaction in step S2 is introduced into an aerobic reactor containing ordinary activated sludge, and microorganisms are added for degradation treatment. The microorganisms are *Roseolario Moraxella salina* (seawater). Rossellomorea aquimaris ZC2402-MT was deposited on May 10, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 30575.

[0042] Example 2: Application of *Moraxella rosenbergii* ZC2402-MT strain in the treatment process of liquor wastewater, including the following steps:

[0043] S1. First, introduce the wastewater into the equalization tank and measure the wastewater quality data, such as COD, pH, ammonia nitrogen, total nitrogen, etc. Control the pH to about 4 to obtain pretreated wastewater.

[0044] S2. The wastewater from S1 is introduced into the anaerobic reactor 1. The anaerobic process produces a large amount of alkalinity, which can be used for pH adjustment to stabilize the pH of the system and reduce the amount of pH adjustment reagent used in the initial stage. The hydraulic retention time is controlled to be 1 day.

[0045] S3. The effluent from the anaerobic reaction in S2 is introduced into aerobic reactor 2 containing ordinary activated sludge, and *Moravir Roselle* ZC2402-MT is added for degradation treatment. The inoculum amount of highly efficient degrading bacteria is 0.1-10%, and in this embodiment, it is 0.1%. A certain mass ratio of trace elements Fe 0-1 mg / L, Cu 0-1 mg / L, Mo 0-1 mg / L, Zn 0-1 mg / L, Co 0-1 mg / L, and Mn are added to the ordinary activated sludge in the reactor. The dosage of each trace element is 0~1 mg / L. In this example, the dosage is 0.1 mg / L. Nutrient elements are added to the aerobic reaction device 2. The ratio of nutrient elements is: NaHCO3 0~1%, MgSO4 0~1%, CaCl2 0~1%, yeast extract 0~0.1%. In this example, NaHCO3 0.5%, MgSO4 0.5%, CaCl2 0.6%, and yeast extract 0.05% are used. The residence time is controlled at 3 days, the temperature is 30-35℃, and the pH is 7.2-7.8.

[0046] S4. Continuously feed the wastewater from S3 into ozone reactor 4, control the residence time to 15 min, and then feed the effluent into BAF reactor, control the residence time to 12 h. Hydrogen peroxide can be added as an ozone catalyst in this step.

[0047] S5. The ozone effluent from S4 is fed into the BAF device to remove the remaining COD and carry out nitrification.

[0048] S6. Pass the wastewater from S5 into the autotrophic denitrification reactor 8 to achieve the goal of extreme denitrification. The effluent will be purified wastewater, and the effluent water quality indicators will be measured.

[0049] Analysis of the effluent and influent water quality shows that the effluent COD removal rate can reach over 99%, with effluent COD less than 30 mg / L, specifically only 20 mg / L; the effluent total nitrogen is 1 mg / L, less than 1.5 mg / L, consistently meeting the Class IV surface water effluent standards. Simultaneously, it reduces the input of solid waste and additional carbon sources and other reagents, lowering costs.

[0050] project COD (mg / L) Ammonia nitrogen (mg / L) Total nitrogen (mg / L) Total phosphorus (mg / L) pH Influent water quality 6477.6 42.66 70.55 22.49 4.51 effluent water quality 20 ＜0.25 1 ＜0.1 7 Limit 30 1.5 1.5 0.3 6-9

[0051] Example 3: Application of *Moraxella rosenbergii* strain ZC2402-MT in the treatment process of liquor wastewater.

[0052] Unlike Example 2, in this example, the inoculum amount of *Roseolario Moraxella salina* ZC2402-MT in step S3 is 2%, and trace elements Fe, Cu, Mo, Zn, Co, and Mn are added to the ordinary activated sludge in the reactor at a dosage of 0.5 mg / L. The nutrient ratio is: NaHCO3 1%, MgSO4 1%, CaCl2 0.6%, and yeast extract 0.1%.

[0053] project COD (mg / L) Ammonia nitrogen (mg / L) Total nitrogen (mg / L) Total phosphorus (mg / L) pH Influent water quality 6477.6 42.66 70.55 22.49 4.51 effluent water quality 16 ＜0.25 1 ＜0.1 7 Limit 30 1.5 1.5 0.3 6-9

[0054] Example 4: Application of *Moraxella rosenbergii* strain ZC2402-MT in the treatment process of soy sauce liquor wastewater.

[0055] Unlike Example 2, in this example, the inoculum amount of *Roseolario Moraxella salina* ZC2402-MT in step S3 is 10%, and trace elements Fe, Cu, Mo, Zn, Co, and Mn are added to the ordinary activated sludge in the reactor at a dosage of 1 mg / L. The nutrient ratio is: 1% NaHCO3, 1% MgSO4, 0.6% CaCl2, and 0.1% yeast extract.

[0056] project COD (mg / L) Ammonia nitrogen (mg / L) Total nitrogen (mg / L) Total phosphorus (mg / L) pH Influent water quality 6477.6 42.66 70.55 22.49 4.51 effluent water quality ＜16 ＜0.25 1 ＜0.1 7 Limit 30 1.5 1.5 0.3 6-9

[0057] Example 5: A process for removing recalcitrant organic matter and performing deep denitrification in liquor wastewater, comprising the following steps:

[0058] S1. First, introduce the wastewater into the equalization tank and measure the wastewater quality data, such as COD, pH, ammonia nitrogen, total nitrogen, etc. Control the pH to about 4 to obtain pretreated wastewater.

[0059] S2. The wastewater from S1 is introduced into the anaerobic reactor. The anaerobic process generates a large amount of alkalinity, which can be used for pH adjustment to stabilize the system pH between 7.2 and 7.8, reducing the amount of initial pH adjustment reagent required. The hydraulic retention time is controlled at 1 day. Then, the wastewater is continuously fed into an aerobic reactor with a conventional activated sludge inoculum of 10%, and 0-1 mg / L of trace elements Fe, Cu, Mo, Zn, Co, and Mn are added. In this example, 0.5 mg / L is used. The retention time is controlled at 3 days. The nutrient ratio is: NaHCO3 1%, MgSO4 1%, CaCl2 0.6%, and yeast extract 0.1%.

[0060] S3. Continuously feed the wastewater from S2 into the ozone reactor, control the residence time to be 15 min, and then feed the effluent into the BAF reactor, control the residence time to be 12 h.

[0061] S4. Pass the wastewater from S3 into the autotrophic denitrification reactor. The effluent will be purified wastewater. Measure the effluent water quality indicators.

[0062] project COD (mg / L) Ammonia nitrogen (mg / L) Total nitrogen (mg / L) Total phosphorus (mg / L) pH Influent water quality 6477.6 42.66 70.55 22.49 4.51 effluent water quality 100 5 10 ＜0.1 7 Limit 30 1.5 1.5 0.3 6-9

[0063] Example 6: A process for removing recalcitrant organic matter and performing deep denitrification in liquor wastewater, comprising the following steps:

[0064] S1. First, introduce the wastewater into the equalization tank and measure the wastewater quality data, such as COD, pH, ammonia nitrogen, total nitrogen, etc. Control the pH to about 4 to obtain pretreated wastewater.

[0065] S2. The wastewater from S1 is introduced into the anaerobic reactor. The anaerobic process generates a large amount of alkalinity, which can be used for pH adjustment to stabilize the system pH and reduce the amount of pH adjustment reagent used initially. The hydraulic retention time is controlled at 1 day. Then, the wastewater is continuously fed into the aerobic reactor with highly efficient degrading bacteria. In this embodiment, 3% of the bacteria are used, and 0-1 mg / L of trace elements Fe, Cu, Mo, Zn, Co, and Mn are added. The retention time is controlled at 3 days.

[0066] S3. Conduct an anaerobic denitrification test on the wastewater in S2, add sodium acetate as a carbon source for denitrification to remove the remaining nitrate nitrogen, and the effluent will be purified wastewater. Measure the effluent water quality indicators.

[0067] project COD (mg / L) Ammonia nitrogen (mg / L) Total nitrogen (mg / L) Total phosphorus (mg / L) pH Influent water quality 6477.6 42.66 70.55 22.49 4.51 effluent water quality 40 12 15 ＜0.1 7 Limit 30 1.5 1.5 0.3 6-9

[0068] Compared with Example 5, Example 2-4 shows that adding the high-efficiency degradation bacteria of the present invention to the aerobic reactor is beneficial to reducing the COD of the effluent and promotes the subsequent purification of various pollutants in the wastewater, so that the effluent can stably meet the standards.

[0069] Compared to Example 6, Examples 2-4 modify the process flow for further treatment of the aerobic effluent. By utilizing advanced ozone oxidation, a BAF reactor, and autotrophic denitrification technology, nitrogen components in the wastewater can be removed more efficiently, while reducing the amount of organic carbon source added, lowering costs, and ensuring that the effluent consistently meets standards. Furthermore, an ozone oxidation catalyst, such as hydrogen peroxide, can be added during ozone oxidation.

[0070] Compared with Examples 3 and 4, Example 2 integrates and optimizes favorable conditions, enabling the removal of various pollutants in the liquor wastewater to the maximum extent, reducing the risk of secondary pollution of the water environment, and has extremely high socio-economic and environmental benefits.

Claims

1. A highly efficient process for degrading recalcitrant organic matter and deeply removing nitrogen from liquor wastewater, characterized in that, The process includes the following steps: S1. Introduce the wastewater into the equalization tank, analyze and measure the water quality, and adjust the influent pH to 3.5-6; S2. The wastewater in the equalization tank of step S1 is introduced into the anaerobic reactor (1) for anaerobic reaction, and the residence time is 0.5-1d; S3. The effluent from the anaerobic reaction in step S2 is introduced into an aerobic reaction device (2) containing ordinary activated sludge, and microorganisms are added for degradation treatment. The microorganisms are *Roseolario Moraxella salina* (seawater). Rossellomorea aquimaris ZC2402-MT was deposited on May 10, 2024, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 30575; S4. After filtration, the aerobic reaction effluent from step S3 enters the ozone reaction device (4) to convert the remaining recalcitrant organic nitrogen into ammonia nitrogen. S5. The ozone effluent from step S4 is fed into the BAF device (6) to remove the remaining COD and carry out nitrification. S6. The BAF effluent from step S5 is fed into the autotrophic denitrification device (8) to achieve the goal of ultimate denitrification.

2. The highly efficient process for degrading recalcitrant organic matter and deeply denitrifying wastewater from soy sauce liquor production as described in claim 1, characterized in that, In step S3, trace elements Fe, Cu, Mo, Zn, Co, and Mn are added to the ordinary activated sludge.

3. The highly efficient process for degrading recalcitrant organic matter and deeply denitrifying wastewater from soy sauce liquor production, as described in claim 2, is characterized in that... The trace element mass ratio is: Fe 0~1mg / L, Cu 0~1mg / L, Mo 0~1mg / L, Zn 0~1mg / L, Co 0~1mg / L, Mn 0~1mg / L.

4. The efficient process for degrading recalcitrant organic matter and deeply denitrifying wastewater from soy sauce liquor as described in claim 1, characterized in that: The mass ratio of the amount of *Roseolario Moraxella salina* ZC2402-MT added to the mass of ordinary activated sludge inoculation is 0.1-10%.

5. The efficient process for degrading recalcitrant organic matter and deeply denitrifying wastewater from soy sauce liquor as described in claim 1, characterized in that: The aerobic reaction device (2) transports wastewater from the bottom. The temperature inside the aerobic reaction device (2) is 33~37℃ and the pH is controlled at 7.2-7.

8.

6. The efficient process for degrading recalcitrant organic matter and deeply denitrifying wastewater from soy sauce liquor production as described in claim 1, characterized in that: Nutrients are added to the aerobic reaction device (2) in the following proportions: NaHCO3 0~1%, MgSO4 0~1%, CaCl2 0~1%, yeast extract 0~0.1%, and pH is maintained between 7.2 and 7.

8.

7. The highly efficient process for degrading recalcitrant organic matter and deeply denitrifying wastewater from soy sauce liquor production, as described in claim 1, is characterized in that... Lightweight packing material (7) is used in the BAF device (6) described in step S5.

8. A strain of bacteria that efficiently degrades recalcitrant organic matter in liquor wastewater, characterized in that, Its classification name is *Roseolium martensii* ( Rossellomorea aquimaris ZC2402-MT was deposited on May 10, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 30575.

9. The microbial agent prepared from the strain that efficiently degrades recalcitrant organic matter in liquor wastewater as described in claim 8.

10. The application of the strain of bacteria that efficiently degrades recalcitrant organic matter in liquor wastewater as described in claim 8 in the liquor wastewater treatment process.

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