Environment-friendly microbial strain and water treatment agent preparation process

Abstract

The invention provides an environment-friendly microbial strain and a water treatment agent preparation process, and relates to the field of water treatment. The environment-friendly microbial strain is prepared from the following raw materials in parts by weight: 20 parts of nitrifying bacteria, 20 parts of denitrifying bacteria, 35 parts of bacillus and 25 parts of EM bacteria, the nitrifying bacteria comprise nitrosomas and nitrobacteria, and the denitrifying bacteria comprise nitrosomas and nitrobacteria. The denitrifying bacteria comprise denitrifying bacilli and sternobacter, the bacillus comprises bacillus subtilis and bacillus licheniformis, and the EM bacteria comprise rhodopseudomonas, lactococcus lactis and saccharomyces cerevisiae. Through flora screening and culture, the culture efficiency is improved, through a fermentation process, fermentation liquor overflow and insufficient dissolved oxygen caused by excessive foam are effectively prevented, the stability of the fermentation process is ensured, through preparation preparation, a good packaging material and a sealing technology, moisture absorption and pollution of a microbial inoculum are prevented, and the quality and safety of a product are ensured.

Description

Technical Field

[0001] The invention relates to the technical field of water treatment, in particular to a preparation process of an environmentally friendly microbial strain and a water treatment agent. Background Art

[0002] With the acceleration of global industrialization, water pollution is becoming increasingly serious. Although traditional physical and chemical water treatment methods (such as precipitation, filtration, adsorption, etc.) can effectively remove some pollutants, they have problems such as high cost, high energy consumption, and easy secondary pollution. At the same time, microbial water treatment technology has gradually become a research hotspot in the field of water treatment due to its advantages such as high efficiency, environmental protection, and sustainability.

[0003] Although the prior art uses microbial agents to achieve good results in water environment management, physical and chemical water treatment methods are prone to secondary pollution, which can easily lead to environmental pollution. Therefore, technicians in this field provide an environmentally friendly microbial strain and a water treatment agent preparation process to solve the problems raised in the above background technology. Summary of the invention

[0004] 1. Technical issues to be resolved

[0005] In view of the deficiencies of the prior art, the present invention provides an environmentally friendly microbial strain and a water treatment agent preparation process, which solves the problem of efficiently degrading pollutants in water and avoiding secondary pollution during sewage treatment.

[0006] (II) Technical solution

[0007] To achieve the above objectives, the present invention is implemented through the following technical scheme: an environmentally friendly microbial strain, including the following raw materials: nitrifying bacteria, denitrifying bacteria, Bacillus and EM bacteria.

[0008] Preferably, the usage ratio of the environmentally friendly microbial strains is: 20 parts of nitrifying bacteria, 20 parts of denitrifying bacteria, 35 parts of Bacillus and 25 parts of EM bacteria.

[0009] Preferably, the nitrifying bacteria genus includes nitrite bacteria and nitrate bacteria, the denitrifying bacteria genus includes denitrifying bacteria and Steinbachia, the Bacillus genus includes Bacillus subtilis and Bacillus licheniformis, and the EM bacteria includes Rhodopseudomonas, Lactococcus lactis and Saccharomyces cerevisiae.

[0010] Preferably, a process for preparing a water treatment agent of environmentally friendly microbial strains comprises the following steps:

[0011] S1. Prepare appropriate culture medium according to the needs of different bacterial species, ensure that the temperature of the culture medium is controlled at 25-37°C and the pH value is 6.5-7.5, place various bacterial communities in their matching culture medium for amplification and culture, use the plate streak method and control the shaking culture speed at 150rpm, and screen out the corresponding bacterial communities by observing the morphology, color and growth rate of the colonies;

[0012] S2, select a stainless steel fermenter, and configure a stirring device, a temperature control system, a pH control system, a dissolved oxygen control system and an exhaust device in the fermenter, sterilize it with high-temperature steam at 121°C for 15 minutes, select glucose, peptone, phosphate and trace elements as culture medium, and put the culture medium into the fermenter after high-temperature sterilization and cooling, add the bacterial flora prepared in S1 into the fermenter according to the proportion, control the growth temperature to 25-37°C, the pH value to 6.5-7.5, stir and ventilate at a speed of 200rpm, and control the dissolved oxygen level to generally between 20% and 40% saturation. According to the fermentation process, add carbon source, nitrogen source and other nutrients in time, and use defoaming agents such as silicone oil and polyether to control the foam in the fermentation process, and let the bacterial flora continue to ferment in the fermenter for 48 hours;

[0013] S3, using a centrifuge at a speed of 5000 rpm to separate the microbial cells in the fermentation broth;

[0014] S4. After obtaining the high-density bacterial colony, the colony is dried by spray drying technology to ensure that the moisture content of the dried bacterial colony is below 5%. The dried bacterial bodies are sealed and packaged and stored in a cool, dry and light-proof environment, and an appropriate amount of trehalose is added thereto to improve the stability of the bacterial colony;

[0015] S5. Add cellulose as a diluent to the bacterial colony and mix them according to the proportion. Use dry granulation technology to make the mixture into a granular preparation. After the preparation is completed, the preparation is sealed and packaged.

[0016] Preferably, the bacterial flora culture in step S1 uses the following formula:

[0017] Calculation of bacterial population specific growth rate:

[0018] Among them, X 1 and X 2 They are t 1 and t 2 The cell concentration at 1 ℃ is used to evaluate the growth rate of bacteria under different culture conditions.

[0019] Preferably, the fermentation process in step S2 uses the following formula:

[0020] Calculation of sterility assurance value:

[0021] Among them, SAL is the sterility assurance value, which is used to evaluate the sterilization effect during the sterilization process;

[0022] Sterilization time calculation:

[0023] Where F is the sterilization value and D is the sterilization time constant at a specific temperature, which is used to determine the time required to reach a specific temperature during the sterilization process.

[0024] Preferably, in steps S1 to S5, waste water, waste gas and waste residue generated during the production process are treated harmlessly, and strict safety operating procedures are formulated to ensure the safety of operators.

[0025] Preferably, after the completion of steps S1 to S5, the bacterial concentration, cell dry weight and other indicators are measured, and the growth curve of the microorganism is drawn to determine whether the fermentation has reached the stable period or the decay period. At the same time, the concentration of the metabolites is analyzed by high performance liquid chromatography and gas chromatography to determine the accumulation of the target product, so as to generally determine the fermentation endpoint.

[0026] (III) Beneficial effects

[0027] The present invention provides a preparation process of environmentally friendly microbial strains and water treatment agents, which have the following beneficial effects:

[0028] 1. Through bacterial screening and cultivation, optimized culture medium and culture conditions (such as temperature and pH value) promote the rapid growth and reproduction of microorganisms, provide sufficient seed liquid for subsequent fermentation, and shaker culture and fermentation tank culture ensure the uniform distribution of microorganisms and sufficient exposure to oxygen, thereby improving the culture efficiency.

[0029] 2. Through the fermentation process, precise control of temperature, pH value, dissolved oxygen and nutrient supply provides optimal conditions for the growth and metabolism of microorganisms, improves the activity of microorganisms and the output of metabolites, and the control of stirring and ventilation ensures the uniform distribution of dissolved oxygen and promotes the uniform growth of microorganisms. At the same time, adding defoaming agents during the fermentation process can effectively prevent the overflow of fermentation liquid and insufficient dissolved oxygen caused by excessive foam, thereby ensuring the stability of the fermentation process.

[0030] 3. Through preparation, the mixture is made into granules using granulation technology, which improves the fluidity and solubility of the product and facilitates the use and transportation of the product. Good packaging materials and sealing technology prevent moisture absorption and contamination of the bacterial agent, ensuring the quality and safety of the product.

[0031] 4. By adding EM bacteria to the bacterial strain, and the EM bacteria include Rhodopseudomonas, Lactococcus lactis and Saccharomyces cerevisiae, Rhodopseudomonas can use light energy to decompose organic matter, which can effectively improve water quality. Lactococcus lactis can inhibit the growth of harmful bacteria, improve the environmental microbial community, and effectively remove odors. Saccharomyces cerevisiae can decompose sugars, purify water quality and promote the decomposition of organic matter. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 It is a schematic diagram of the process of the present invention;

[0033] Figure 2 A schematic diagram of the process of sampling the bacterial flora of the present invention;

[0034] Figure 3 Schematic diagram of the process for preparing the preparation of the present invention. DETAILED DESCRIPTION

[0035] 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.

[0036] Embodiment 1:

[0037] like Figure 1-3 As shown, an embodiment of the present invention provides an environmentally friendly microbial strain, including the following raw materials: nitrifying bacteria, denitrifying bacteria, Bacillus and EM bacteria, the usage ratio of the environmentally friendly microbial strains is: 20 parts of nitrifying bacteria, 20 parts of denitrifying bacteria, 35 parts of Bacillus and 25 parts of EM bacteria, the nitrifying bacteria include nitrite bacteria and nitrate bacteria, the denitrifying bacteria include denitrifying bacteria and Steinbachia, the Bacillus includes Bacillus subtilis and Bacillus licheniformis, the EM bacteria include Rhodopseudomonas, Lactococcus lactis and Saccharomyces cerevisiae, by adding EM bacteria to the strain, and the EM bacteria include Rhodopseudomonas, Lactococcus lactis and Saccharomyces cerevisiae, Rhodopseudomonas can use light energy to decompose organic matter, which can effectively improve water quality, Lactococcus lactis can inhibit the growth of harmful bacteria, improve environmental microbial communities, and effectively remove odors, and Saccharomyces cerevisiae can decompose sugars, purify water quality and promote the decomposition of organic matter.

[0038] A process for preparing a water treatment agent using environmentally friendly microbial strains comprises the following steps:

[0039] S1. According to the needs of different bacterial species, prepare appropriate culture medium, ensure that the temperature of the culture medium is controlled at 25-37°C and the pH value is 6.5-7.5, place various bacterial communities in their matching culture medium for amplification and culture, use the plate streak method and control the shaking speed at 150rpm, and screen out the corresponding bacterial communities by observing the morphology, color and growth rate of the colonies. Through bacterial community screening and culture, optimized culture medium and culture conditions (such as temperature and pH value) promote the rapid growth and reproduction of microorganisms, provide sufficient seed liquid for subsequent fermentation, and shaker culture and fermentation tank culture ensure the uniform distribution of microorganisms and sufficient contact with oxygen, thereby improving the culture efficiency.

[0040] S2. Select a stainless steel fermenter, and equip the fermenter with a stirring device, a temperature control system, a pH control system, a dissolved oxygen control system and an exhaust device. Sterilize with high-temperature steam at 121°C for 15 minutes. Select glucose, peptone, phosphate and trace elements as culture media, and put the culture media into the fermenter after high-temperature sterilization and cooling. Add the bacterial flora prepared in S1 to the fermenter in proportion, control the growth temperature to 25-37°C, the pH value to 6.5-7.5, stir and ventilate at a speed of 200rpm, and control the dissolved oxygen level to 20% to 40% saturation. According to the fermentation process, carbon source, nitrogen source and other nutrients are added in time, and defoaming agents such as silicone oil and polyether are used to control the foam in the fermentation process. The bacterial community is allowed to ferment continuously in the fermentation tank for 48 hours. Through the fermentation process, precise control of temperature, pH value, dissolved oxygen and nutrient supply provides the best conditions for the growth and metabolism of microorganisms, improves the activity of microorganisms and the output of metabolites, and the control of stirring and ventilation ensures the uniform distribution of dissolved oxygen and promotes the uniform growth of microorganisms. At the same time, adding defoaming agents during the fermentation process can effectively prevent the overflow of fermentation liquid and insufficient dissolved oxygen caused by excessive foam, thereby ensuring the stability of the fermentation process.

[0041] S3, using a centrifuge at a speed of 5000 rpm to separate the microbial cells in the fermentation broth;

[0042] S4. After obtaining a high-density bacterial colony, the colony is dried using spray drying technology to ensure that the moisture content of the dried bacterial colony is below 5%. The dried bacterial bodies are sealed and packaged and stored in a cool, dry, light-proof environment, and an appropriate amount of trehalose is added thereto to improve the stability of the bacterial colony.

[0043] S5. Add cellulose as a diluent to the bacterial colony and mix them in proportion. Use dry granulation technology to make the mixture into a granular preparation. After the preparation is completed, the preparation is sealed and packaged. Through the preparation preparation, the mixture is made into granules using dry granulation technology, which improves the fluidity and solubility of the product and facilitates the use and transportation of the product. Good packaging materials and sealing technology prevent the bacterial agent from absorbing moisture and contamination, thereby ensuring the quality and safety of the product.

[0044] The following formula is used for bacterial colony culture in step S1:

[0045] Calculation of bacterial population specific growth rate:

[0046] Among them, X 1 and X 2 They are t 1 and t 2 The cell concentration at 1 ℃ is used to evaluate the growth rate of bacteria under different culture conditions.

[0047] The following formula is used in the fermentation process in step S2:

[0048] Calculation of sterility assurance value:

[0049] Among them, SAL is the sterility assurance value, which is used to evaluate the sterilization effect during the sterilization process;

[0050] Sterilization time calculation:

[0051] Where F is the sterilization value and D is the sterilization time constant at a specific temperature, which is used to determine the time required to reach a specific temperature during the sterilization process.

[0052] In steps S1 to S5, the waste water, waste gas and waste residue generated in the production process are treated harmlessly, and strict safety operating procedures are formulated to ensure the safety of operators. After the completion of steps S1 to S5, the bacterial concentration, cell dry weight and other indicators are measured, and the growth curve of the microorganism is drawn to determine whether the fermentation has reached the stable period or the decay period. At the same time, high-performance liquid chromatography and gas chromatography are used to analyze the concentration of metabolites to determine the accumulation of target products, so as to determine the overall end point of the fermentation.

[0053] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An environmentally friendly microbial strain, characterized by: Including the following ingredients: Nitrifying bacteria, denitrifying bacteria, Bacillus and EM bacteria.

2. The environmentally friendly microbial strain according to claim 1, characterized in that: The usage ratio of the environmentally friendly microbial strains is: 20 parts of nitrifying bacteria, 20 parts of denitrifying bacteria, 35 parts of Bacillus and 25 parts of EM bacteria.

3. The environmentally friendly microbial strain according to claim 1, characterized in that: The nitrifying bacteria genus includes nitrite bacteria and nitrate bacteria, the denitrifying bacteria genus includes denitrifying bacteria and Steinbachia, the Bacillus genus includes Bacillus subtilis and Bacillus licheniformis, and the EM bacteria includes Rhodopseudomonas, Lactococcus lactis and Saccharomyces cerevisiae.

4. The process for preparing a water treatment agent using environmentally friendly microbial strains according to claim 1, characterized in that: The following steps are involved: S1. Prepare appropriate culture medium according to the needs of different bacterial species, ensure that the temperature of the culture medium is controlled at 25-37°C and the pH value is 6.5-7.5, place various bacterial communities in their matching culture medium for amplification and culture, use the plate streak method and control the shaking culture speed at 150rpm, and screen out the corresponding bacterial communities by observing the morphology, color and growth rate of the colonies; S2, select a stainless steel fermenter, and configure a stirring device, a temperature control system, a pH control system, a dissolved oxygen control system and an exhaust device in the fermenter, sterilize it with high-temperature steam at 121°C for 15 minutes, select glucose, peptone, phosphate and trace elements as culture medium, and put the culture medium into the fermenter after high-temperature sterilization and cooling, add the bacterial flora prepared in S1 into the fermenter according to the proportion, control the growth temperature to 25-37°C, the pH value to 6.5-7.5, stir and ventilate at a speed of 200rpm, and control the dissolved oxygen level to generally between 20% and 40% saturation. According to the fermentation process, add carbon source, nitrogen source and other nutrients in time, and use defoaming agents such as silicone oil and polyether to control the foam in the fermentation process, and let the bacterial flora continue to ferment in the fermenter for 48 hours; S3, using a centrifuge at a speed of 5000 rpm to separate the microbial cells in the fermentation broth; S4. After obtaining the high-density bacterial colony, the colony is dried by spray drying technology to ensure that the moisture content of the dried bacterial colony is below 5%. The dried bacterial bodies are sealed and packaged and stored in a cool, dry and light-proof environment, and an appropriate amount of trehalose is added thereto to improve the stability of the bacterial colony; S5. Add cellulose as a diluent to the bacterial colony and mix them according to the proportion. Use dry granulation technology to make the mixture into a granular preparation. After the preparation is completed, the preparation is sealed and packaged.

5. The process for preparing a water treatment agent using environmentally friendly microbial strains according to claim 1, characterized in that: The following formula is used for the bacterial colony culture in step S1: Calculation of bacterial population specific growth rate: Among them, X1 and X2 are the cell concentrations at t1 and t2, respectively, which are used to evaluate the growth rate of bacteria under different culture conditions.

6. The process for preparing a water treatment agent using environmentally friendly microbial strains according to claim 1, characterized in that: The fermentation process in step S2 uses the following formula: Calculation of sterility assurance value: Among them, SAL is the sterility assurance value, which is used to evaluate the sterilization effect during the sterilization process; Sterilization time calculation: Where F is the sterilization value and D is the sterilization time constant at a specific temperature, which is used to determine the time required to reach a specific temperature during the sterilization process.

7. The process for preparing a water treatment agent using environmentally friendly microbial strains according to claim 1, characterized in that: In the steps S1 to S5, the waste water, waste gas and waste residue generated in the production process are treated harmlessly, and strict safety operating procedures are formulated to ensure the safety of operators.

8. The process for preparing a water treatment agent using environmentally friendly microbial strains according to claim 1, characterized in that: After the steps S1 to S5 are completed, the bacterial concentration, cell dry weight and other indicators are measured and the growth curve of the microorganism is drawn to determine whether the fermentation has reached the stable period or the decay period. At the same time, the concentration of the metabolites is analyzed by high performance liquid chromatography and gas chromatography to determine the accumulation of the target product, so as to generally determine the fermentation endpoint.

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