Preparation method of microbial agent for waste gas treatment

Abstract

The invention discloses a preparation method of a microbial agent for waste gas treatment, and particularly relates to the technical field of waste gas treatment, the specific preparation method comprises the following steps: step 1, strain selection; step 2, genetic modification; 3, preparing a culture medium; step 4, inoculation and primary fermentation; step 5, secondary fermentation; step 6, separating fermentation liquor; step 7, preparation preparation; step 8, quality detection; and step 9, packaging and storing. According to the invention, genetic modification is carried out on the selected microbial strain by using a gene editing technology, so that the microbial inoculum has high-efficiency degradation capability on specific pollutants in waste gas, the content of harmful substances in the waste gas can be effectively reduced, the improvement of the environmental treatment efficiency is also facilitated, and through the secondary fermentation process, the environmental pollution is reduced. And specific enzymes and beneficial microorganisms are added, so that the degradation capability of the microbial agent and the adaptability to different environments are enhanced, and the microbial agent is more stable in complex and changeable waste gas treatment environments.

Description

Technical Field

[0001] The present invention relates to the technical field of waste gas treatment, and more specifically, to a method for preparing a microbial agent for waste gas treatment. Background Art

[0002] Waste gas treatment, also known as waste gas purification, mainly refers to the work of treating industrial waste gas such as dust particles, flue gas, odorous gas, and toxic and harmful gas generated in industrial places. Common waste gas purification includes factory smoke waste gas purification, workshop dust waste gas purification, organic waste gas purification, waste gas odor purification, acid and alkali waste gas purification, chemical waste gas purification, etc. With the rapid development of industrialization, the environmental pollution caused by waste gas emissions is becoming more and more serious. The harmful substances in the waste gas pose a serious threat to the atmospheric environment and human health. Therefore, it is imperative to develop efficient and environmentally friendly waste gas treatment technology.

[0003] As a biological treatment technology, microbial agents play an important role in waste gas treatment. Although traditional microbial agents can degrade pollutants in waste gas to a certain extent, they often have problems such as low degradation efficiency and poor adaptability. In order to overcome these challenges, researchers are constantly exploring new production methods to improve the performance of microbial agents.

[0004] After searching, the Chinese patent with application number CN202010228796.1 discloses a microbial deodorizing agent and its preparation method and waste gas treatment process, including: enriching the dominant bacterial community after degrading the kitchen waste, separating and comparing the strains, selecting safe dominant strains for combination and compounding for efficient treatment of waste gas generated by the microbial metabolism of organic waste. The exhaust gas composition after treatment by the invention method fully meets the emission requirements, and no odor is released during the entire process of organic waste treatment, which is environmentally friendly.

[0005] In summary, the above patents show that the existing methods for preparing microbial agents that can be used for waste gas treatment have unsatisfactory degradation capabilities for specific pollutants in waste gas during actual use, and are unable to effectively reduce the content of harmful substances in waste gas, resulting in low environmental governance efficiency. In addition, the degradation capabilities and adaptability of the agents to different environments are poor, resulting in the agents being unstable in complex and changeable waste gas treatment environments. Summary of the invention

[0006] In order to overcome the above-mentioned defects of the prior art, the present invention provides a method for preparing a microbial agent for waste gas treatment.

[0007] To achieve the above object, the present invention provides the following technical solution: a method for preparing a microbial agent for waste gas treatment, the specific steps of the preparation method are as follows:

[0008] Step 1: Bacteria selection: According to the specific requirements of waste gas treatment, select microbial strains that can degrade specific pollutants in the waste gas;

[0009] Step 2: Genetic modification: Genetically modify the selected bacteria using gene editing technology to improve their degradation efficiency and adaptability to specific pollutants in exhaust gas;

[0010] Step 3: Culture medium preparation: prepare a culture medium suitable for the growth of the selected bacterial species;

[0011] Step 4: Inoculation and primary fermentation: Under sterile conditions, the selected bacteria are inoculated into the prepared sterile culture medium for primary fermentation;

[0012] Step 5: Secondary fermentation: After the first fermentation is completed, specific enzymes and beneficial microorganisms are added for secondary fermentation. This step can enhance the degradation ability and adaptability of the microbial agent.

[0013] Step 6, separation of fermentation liquid: separating the liquid after secondary fermentation from the bacterial cells by filter pressing or centrifugation, and collecting the bacterial cells;

[0014] Step 7: Preparation of preparation: the collected bacteria are mixed with an appropriate amount of carrier material, and the desired dosage form is prepared through drying and granulation steps. The formed dosage form includes powder or granule. During this process, the stability and activity of the bacterial agent must be ensured;

[0015] Step 8: Quality inspection: Conduct quality inspection on the finished bacterial agent;

[0016] Step 9. Packaging and storage: Package and store qualified microbial agents.

[0017] As a further improvement of the technical solution of the present invention, the microbial strain selected in the step 1 includes any one of bacteria or fungi with high-efficiency degradation ability.

[0018] As a further improvement of the technical solution of the present invention, the method of genetically modifying the selected bacterial species in step 2 includes using the CRISPR-Cas9 gene editing tool to direct the metabolic pathway of the bacterial species so that it can more efficiently transform and degrade pollutants.

[0019] As a further improvement of the technical solution of the present invention, the culture medium in step three contains essential components of carbon source, nitrogen source, inorganic salts and growth factors to support the growth and reproduction of microorganisms, wherein the carbon source is preferably sucrose, and the nitrogen source is preferably yeast extract or peptone.

[0020] As a further improvement of the technical solution of the present invention, in step 4, the fermentation temperature is maintained at 25-30° C. and the fermentation time is 30-35 days.

[0021] As a further improvement of the technical solution of the present invention, the enzyme added in step five includes any one of Aspergillus, cellulase or protease, the added beneficial microorganisms are preferably actinomycetes, the temperature of the secondary fermentation is maintained at 30-35°C, and the fermentation time is 7 days.

[0022] As a further improvement of the technical solution of the present invention, the filter pressing operation steps in step six are as follows:

[0023] S1. Prepare filter press equipment: select suitable filter plates and filter cloths to ensure that they can withstand the pressure and corrosiveness of the fermentation liquid;

[0024] S2. Apply pressure: Pour the fermentation liquid into the filter press equipment, and then apply appropriate pressure to force the liquid to pass through the filter cloth;

[0025] S3, collecting filtrate and bacteria: the filter cloth will retain the bacteria and other solid particles, while the filtrate flows out through the filter cloth, and the filtrate and the bacteria retained on the filter cloth are collected separately;

[0026] S4. Cleaning and drying: If necessary, the filter cloth can be cleaned to remove residual bacteria and impurities, and then the retained bacteria are dried.

[0027] As a further improvement of the technical solution of the present invention, the centrifugation method in step six is ​​operated as follows:

[0028] S1. Prepare the centrifuge: Select a suitable centrifuge according to the amount of fermentation liquid. If the amount is small, a refrigerated centrifuge can be used, and if the amount is large, a tubular centrifuge can be used;

[0029] S2. Sample loading and balance: Pour the fermentation liquid evenly into the centrifuge tube and ensure that the centrifuge tube is placed in a balanced position in the centrifuge;

[0030] S3. Set centrifugation parameters: set the centrifugation speed to 5000-10000 rpm and the centrifugation time to 30-60 min;

[0031] S4, start the centrifuge: start the centrifuge and let it run at the set speed and time;

[0032] S5. Collect bacteria: After centrifugation, turn off the centrifuge and take out the centrifuge tube. At this time, the bacteria will settle at the bottom of the centrifuge tube. You can carefully pour out the supernatant and leave the bacteria for subsequent processing.

[0033] As a further improvement of the technical solution of the present invention, in step eight, the quality inspection of the finished microbial agent includes the number of effective live bacteria, the number of foreign bacteria and safety indicators to ensure that the product quality meets the standards.

[0034] As a further improvement of the technical solution of the present invention, the packaging of the qualified bacterial agent in step nine includes packaging in plastic bottles or aluminum foil bags, and the packaged bacterial agent is stored in a dry, light-proof and ventilated place, and the temperature is maintained at 15-25°C.

[0035] Beneficial effects of the present invention:

[0036] The present invention utilizes gene editing technology to genetically modify the selected microbial strains, so that the bacterial agent has the ability to efficiently degrade specific pollutants in the waste gas, which can not only effectively reduce the content of harmful substances in the waste gas, but also help to improve the efficiency of environmental governance. Through the secondary fermentation process, specific enzymes and beneficial microorganisms are added to enhance the degradation ability of the bacterial agent and its adaptability to different environments, making the bacterial agent more stable in the complex and changeable waste gas treatment environment. In summary, the microbial agent production method designed by the present invention shows the characteristics of high efficiency, stability, safety and convenience in technical effects, and provides an effective and practical solution for the field of waste gas treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The present invention is a flow chart of the method for preparing the microbial agent. DETAILED DESCRIPTION

[0038] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] As attached Figure 1 The process shown provides a method for preparing a microbial agent for waste gas treatment, and the specific steps of the preparation method are as follows:

[0040] Step 1: Bacteria selection: According to the specific requirements of waste gas treatment (refer to the "Comprehensive Emission Standard of Air Pollutants" (GB16297-1996), and combined with the actual treatment requirements), select microbial strains that can degrade specific pollutants in the waste gas;

[0041] Step 2: Genetic modification: Genetically modify the selected bacteria using gene editing technology (increasing or enhancing new traits of microorganisms in treating harmful substances in waste gas) to improve their degradation efficiency and adaptability to specific pollutants in waste gas;

[0042] Step 3: Preparation of culture medium: According to the bacterial strain selected and transformed in step 2, a culture medium suitable for the growth conditions of the bacterial strain is prepared;

[0043] Step 4: Inoculation and primary fermentation: Under sterile conditions, the selected and modified strains are inoculated into the sterile culture medium prepared in step 3 for primary fermentation;

[0044] Step 5: Secondary fermentation: After the first fermentation is completed, specific enzymes and beneficial microorganisms are added for secondary fermentation. This step can enhance the degradation ability and adaptability of the microbial agent.

[0045] Step 6, separation of fermentation liquid: separating the bacterial cells in the fermentation liquid by filter pressing or centrifugation, and collecting the bacterial cells;

[0046] Step 7: Preparation of preparation: the collected bacteria are mixed with an appropriate amount of carrier material, and then dried and granulated to form the desired dosage form, including powder or granules. During this process, the stability and activity of the bacterial agent must be ensured;

[0047] Step 8: Quality inspection: Conduct quality inspection on the finished bacterial agent;

[0048] Step 9. Packaging and storage: Package and store qualified microbial agents.

[0049] In an embodiment of the present invention, the microbial strain selected in the above step 1 includes any one of bacteria or fungi with high-efficiency degradation ability.

[0050] In addition, the method of genetically modifying the selected bacterial strain in the above step 2 includes using the CRISPR-Cas9 gene editing tool to target the metabolic pathway of the bacterial strain so that it can more efficiently transform and degrade pollutants.

[0051] In addition, the culture medium in the above step three contains essential components of carbon source, nitrogen source, inorganic salts and growth factors, which are mainly used to support the growth and reproduction of microorganisms. Among them, the carbon source is preferably sucrose, and the nitrogen source is preferably yeast extract or peptone.

[0052] In an embodiment of the present invention, the fermentation temperature in step 4 is controlled at 25-30° C., and the fermentation time is preferably 30-35 days.

[0053] In an embodiment of the present invention, the enzyme added in the above step five includes any one of Aspergillus, cellulase or protease, and the added beneficial microorganisms are preferably actinomycetes, and the temperature of the secondary fermentation is controlled at 30-35°C, and the fermentation time is preferably 7 days.

[0054] The filter pressing operation in step 6 of the above embodiment includes the following steps:

[0055] S1. Prepare filter press equipment: select suitable filter plates and filter cloths to ensure that they can withstand the pressure and corrosiveness of the fermentation liquid;

[0056] S2. Apply pressure: Pour the fermentation liquid into the filter press equipment, and then apply appropriate pressure to force the liquid to pass through the filter cloth;

[0057] S3, collecting filtrate and bacteria: the filter cloth will retain the bacteria and other solid particles, while the filtrate flows out through the filter cloth, and the filtrate and the bacteria retained on the filter cloth are collected separately;

[0058] S4. Cleaning and drying: If necessary, the filter cloth can be cleaned to remove residual bacteria and impurities, and then the retained bacteria are dried.

[0059] In this embodiment, the centrifugation method in step 6 above includes the following steps:

[0060] A. Prepare the centrifuge: Choose a suitable centrifuge according to the amount of fermentation liquid. If the amount is small, a refrigerated centrifuge can be used, and if the amount is large, a tubular centrifuge can be used;

[0061] B. Sample loading and balance: Pour the fermentation liquid evenly into the centrifuge tube and ensure that the centrifuge tube is placed in a balanced position in the centrifuge;

[0062] C. Set centrifugal parameters: set the centrifugal speed to 5000-10000rpm and the centrifugal time to 30-60min;

[0063] D. Start the centrifuge: Start the centrifuge and let it run at the set speed and time;

[0064] F. Collecting bacteria: After centrifugation, turn off the centrifuge and take out the centrifuge tube. At this time, the bacteria will settle at the bottom of the centrifuge tube. You can carefully pour out the supernatant and leave the bacteria for subsequent processing.

[0065] In addition, in step eight of the above embodiment, the quality of the finished microbial agent is tested, including the number of effective live bacteria, the number of foreign bacteria and safety indicators, to ensure that the product quality meets the standards.

[0066] Furthermore, the packaging of the qualified bacterial agent in step nine includes packaging in plastic bottles or aluminum foil bags, and the packaged bacterial agent is stored in a dry, light-proof and ventilated place, and the temperature is maintained at 15-25°C.

[0067] Working principle: The present invention designs a method for preparing a microbial agent for waste gas treatment. First, according to the specific needs of waste gas treatment, a microbial strain capable of degrading specific pollutants in the waste gas is selected. The selected microbial strain includes any one of bacteria or fungi with high-efficiency degradation ability;

[0068] Secondly, using gene editing technology, the selected bacteria are genetically modified to improve their degradation efficiency and adaptability to specific pollutants in waste gas. For example, the CRISPR-Cas9 gene editing tool is used to modify the metabolic pathways of bacteria so that they can more efficiently transform and degrade pollutants. Through carefully selected microbial strains and genetic modification using gene editing technology, the bacterial agents have the ability to efficiently degrade specific pollutants in waste gas, which can not only effectively reduce the content of harmful substances in waste gas, but also help improve the efficiency of environmental governance.

[0069] Then, a culture medium suitable for the growth of the selected bacterial species is prepared, wherein the culture medium contains carbon source, nitrogen source, inorganic salt and necessary growth factor components to support the growth and reproduction of the microorganism, wherein the carbon source is preferably sucrose, and the nitrogen source is preferably yeast extract or peptone;

[0070] Further, under sterile conditions, the selected strains are inoculated into the prepared sterile culture medium for a fermentation, the fermentation temperature is maintained at 25-30° C., and the fermentation time is 30-35 days;

[0071] Furthermore, after the completion of the first fermentation, specific enzymes and beneficial microorganisms are added to carry out a second fermentation. This step can enhance the degradation ability and adaptability of the microbial agent. The added enzyme includes any one of Aspergillus, cellulase or protease. The added beneficial microorganism is preferably actinomycetes. The temperature of the second fermentation is maintained at 30-35°C and the fermentation time is 7 days. Through the second fermentation process, specific enzymes and beneficial microorganisms are added, which further enhances the degradation ability and adaptability of the microbial agent to different environments. This enhanced treatment makes the microbial agent more stable in the complex and changeable waste gas treatment environment.

[0072] Then, the liquid after secondary fermentation is separated from the bacterial cells by filtration or centrifugation to collect the bacterial cells; wherein the filtration operation steps are as described in the above implementation contents as follows:

[0073] S1. Prepare filter press equipment: select suitable filter plates and filter cloths to ensure that they can withstand the pressure and corrosiveness of the fermentation liquid;

[0074] S2. Apply pressure: Pour the fermentation liquid into the filter press equipment, and then apply appropriate pressure to force the liquid to pass through the filter cloth;

[0075] S3, collecting filtrate and bacteria: the filter cloth will retain the bacteria and other solid particles, while the filtrate flows out through the filter cloth, and the filtrate and the bacteria retained on the filter cloth are collected separately;

[0076] S4. Cleaning and drying: If necessary, the filter cloth can be cleaned to remove residual bacteria and impurities, and then the retained bacteria are dried.

[0077] The centrifugation method operation steps are as follows:

[0078] A. Prepare the centrifuge: Choose a suitable centrifuge according to the amount of fermentation liquid. If the amount is small, a refrigerated centrifuge can be used, and if the amount is large, a tubular centrifuge can be used;

[0079] B. Sample loading and balance: Pour the fermentation liquid evenly into the centrifuge tube and ensure that the centrifuge tube is placed in a balanced position in the centrifuge;

[0080] C. Set centrifugal parameters: set the centrifugal speed to 5000-10000rpm and the centrifugal time to 30-60min;

[0081] D. Start the centrifuge: Start the centrifuge and let it run at the set speed and time;

[0082] F. Collecting bacteria: After centrifugation, turn off the centrifuge and take out the centrifuge tube. At this time, the bacteria will settle at the bottom of the centrifuge tube. You can carefully pour out the supernatant and leave the bacteria for subsequent processing.

[0083] Then, the collected bacteria are mixed with an appropriate amount of carrier material, and the desired dosage form is prepared through drying and granulation steps. The formed dosage form includes powder or granule. In this process, the stability and activity of the microbial agent must be ensured. The carrier materials include wood particles, coal slag, organic carriers (such as bran, straw powder and distiller's grains powder, etc.) and inorganic carriers (such as diatomaceous earth and zeolite powder, etc.). In the preparation process, the stability and activity of the microbial agent are ensured by optimizing the selection of carrier materials and processing steps, which means that the microbial agent can maintain its efficacy during storage and use, thereby providing a more reliable treatment effect;

[0084] Furthermore, the finished microbial agent is subjected to quality inspection, including the number of effective live bacteria, number of foreign bacteria and safety indicators. By conducting comprehensive quality inspection on the finished microbial agent, including the number of effective live bacteria, number of foreign bacteria and safety indicators, it is ensured that the quality of the product meets the industry standards, providing users with safe and reliable products;

[0085] Finally, the qualified microbial agents are packaged and stored, including in plastic bottles or aluminum foil bags. The packaged microbial agents are stored in a dry, light-proof and ventilated place and the temperature is maintained at 15-25°C. The use of appropriate packaging and storage conditions not only facilitates the transportation and distribution of the product, but also extends the shelf life of the product, allowing it to maintain good performance in different regions and environmental conditions.

[0086] In summary, the present invention uses gene editing technology to genetically modify the selected microbial strains, so that the bacterial agent has the ability to efficiently degrade specific pollutants in the waste gas, which can not only effectively reduce the content of harmful substances in the waste gas, but also help to improve the efficiency of environmental governance. Through the secondary fermentation process, specific enzymes and beneficial microorganisms are added to enhance the degradation ability of the bacterial agent and its adaptability to different environments, making the bacterial agent more stable in the complex and changeable waste gas treatment environment.

[0087] 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 principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a microbial agent for waste gas treatment, characterized in that: The specific production steps are as follows: Step 1: Bacteria selection: According to the specific requirements of waste gas treatment, select microbial strains that can degrade specific pollutants in the waste gas; Step 2: Genetic modification: Genetically modify the selected strains using gene editing technology; Step 3: Culture medium preparation: prepare a culture medium suitable for the growth of the selected bacterial species; Step 4: Inoculation and primary fermentation: Under sterile conditions, the selected bacteria are inoculated into the prepared sterile culture medium for primary fermentation; Step 5: Secondary fermentation: After the first fermentation is completed, specific enzymes and beneficial microorganisms are added for secondary fermentation; Step 6, separation of fermentation liquid: separating the liquid after secondary fermentation from the bacterial cells by filter pressing or centrifugation, and collecting the bacterial cells; Step 7, preparation of preparation: the collected bacteria are mixed with an appropriate amount of carrier material, and the desired dosage form is prepared through drying and granulation steps, and the formed dosage form includes powder or granules; Step 8: Quality inspection: Conduct quality inspection on the finished bacterial agent; Step 9. Packaging and storage: Package and store qualified microbial agents.

2. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: The microbial strain selected in step 1 includes any one of bacteria or fungi with high-efficiency degradation ability.

3. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: The method of genetically modifying the selected bacterial species in step 2 includes using the CR ISPR-Cas9 gene editing tool to direct the modification of the metabolic pathway of the bacterial species.

4. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: In step 3, the culture medium contains essential components of carbon source, nitrogen source, inorganic salt and growth factor to support the growth and reproduction of microorganisms, wherein the carbon source is preferably sucrose, and the nitrogen source is preferably yeast extract or peptone.

5. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: In step 4, the fermentation temperature is maintained at 25-30° C. and the fermentation time is 30-35 days.

6. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: The enzyme added in step 5 includes any one of Aspergillus, cellulase or protease, and the added beneficial microorganisms are preferably actinomycetes. The temperature of the secondary fermentation is maintained at 30-35° C. and the fermentation time is 7 days.

7. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: The filter pressing operation steps in step 6 are as follows: S1. Prepare filter press equipment: select suitable filter plates and filter cloths; S2. Apply pressure: Pour the fermentation liquid into the filter press equipment, and then apply appropriate pressure to force the liquid to pass through the filter cloth; S3, collecting filtrate and bacteria: the filter cloth will retain the bacteria and other solid particles, while the filtrate flows out through the filter cloth, and the filtrate and the bacteria retained on the filter cloth are collected separately; S4, cleaning and drying: as needed, the filter cloth is cleaned to remove residual bacteria and impurities, and then the retained bacteria are dried.

8. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: The centrifugation method in step 6 is as follows: S1. Prepare the centrifuge: Select a suitable centrifuge according to the amount of fermentation liquid. If the amount is small, a refrigerated centrifuge can be used, and if the amount is large, a tubular centrifuge can be used; S2, sample loading and balance: Pour the fermentation liquid evenly into the centrifuge tube and place the centrifuge tube in a balanced position in the centrifuge; S3. Set centrifugation parameters: set the centrifugation speed to 5000-10000 rpm and the centrifugation time to 30-60 min; S4, start the centrifuge: start the centrifuge and let it run at the set speed and time; S5. Collecting bacteria: After centrifugation, turn off the centrifuge and take out the centrifuge tube. At this time, the bacteria will settle at the bottom of the centrifuge tube. Pour out the supernatant and leave the bacteria for subsequent processing.

9. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: In step eight, the quality inspection of the finished microbial agent includes the number of effective live bacteria, the number of foreign bacteria and safety indicators.

10. The method for preparing a microbial agent for waste gas treatment according to claim 1, characterized in that: The packaging of the qualified bacterial agent in step nine includes packaging in plastic bottles or aluminum foil bags. The packaged bacterial agent is stored in a dry, light-proof and ventilated place and maintained at a temperature of 15-25°C.

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