CN118324274A - Preparation method of microbial reinforced flocculant - Google Patents
Preparation method of microbial reinforced flocculant Download PDFInfo
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- CN118324274A CN118324274A CN202410349311.2A CN202410349311A CN118324274A CN 118324274 A CN118324274 A CN 118324274A CN 202410349311 A CN202410349311 A CN 202410349311A CN 118324274 A CN118324274 A CN 118324274A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 102000004190 Enzymes Human genes 0.000 claims abstract description 18
- 108090000790 Enzymes Proteins 0.000 claims abstract description 18
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 15
- 239000008103 glucose Substances 0.000 claims abstract description 15
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 238000005728 strengthening Methods 0.000 claims abstract description 12
- 150000004676 glycans Chemical class 0.000 claims abstract description 11
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 11
- 239000005017 polysaccharide Substances 0.000 claims abstract description 11
- 229920002472 Starch Polymers 0.000 claims abstract description 10
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- 235000019698 starch Nutrition 0.000 claims abstract description 10
- 229920001285 xanthan gum Polymers 0.000 claims abstract description 10
- 239000000230 xanthan gum Substances 0.000 claims abstract description 10
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- 235000010493 xanthan gum Nutrition 0.000 claims abstract description 10
- 229920001218 Pullulan Polymers 0.000 claims abstract description 8
- 239000004373 Pullulan Substances 0.000 claims abstract description 8
- 238000009629 microbiological culture Methods 0.000 claims abstract description 8
- 235000019423 pullulan Nutrition 0.000 claims abstract description 8
- 229920000310 Alpha glucan Polymers 0.000 claims abstract description 6
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 5
- 239000012744 reinforcing agent Substances 0.000 claims abstract description 5
- -1 levoglucosides Substances 0.000 claims abstract description 4
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- 230000001804 emulsifying effect Effects 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 11
- 238000000855 fermentation Methods 0.000 claims description 10
- 230000004151 fermentation Effects 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 10
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- 238000007710 freezing Methods 0.000 claims description 6
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- 239000012528 membrane Substances 0.000 claims description 6
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- 239000000701 coagulant Substances 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 239000011148 porous material Substances 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 239000013049 sediment Substances 0.000 claims description 3
- 238000004062 sedimentation Methods 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 244000005700 microbiome Species 0.000 abstract description 23
- 230000000694 effects Effects 0.000 abstract description 16
- 238000005189 flocculation Methods 0.000 abstract description 10
- 230000016615 flocculation Effects 0.000 abstract description 10
- 150000002500 ions Chemical class 0.000 abstract description 8
- 229910001415 sodium ion Inorganic materials 0.000 abstract description 7
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 abstract description 6
- 229910001424 calcium ion Inorganic materials 0.000 abstract description 6
- 125000000524 functional group Chemical group 0.000 abstract description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 abstract description 3
- 210000000170 cell membrane Anatomy 0.000 abstract description 3
- 210000002421 cell wall Anatomy 0.000 abstract description 3
- 230000003993 interaction Effects 0.000 abstract description 2
- 238000012545 processing Methods 0.000 abstract description 2
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- 238000004519 manufacturing process Methods 0.000 description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 5
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the technical field of flocculant processing, and discloses a preparation method of a microbial enhanced flocculant, wherein the preparation method of the microbial enhanced flocculant comprises the following steps: preparing a microbial culture medium: selecting glucose, xanthan gum, levoglucosides, pullulan alpha-glucan and soluble starch to form a polysaccharide culture medium; and (3) preparing a reinforcing agent: naC l and CaCl 2 were selected to be configured as strengthening solutions. According to the preparation method of the microbial reinforced flocculant, the sodium ions (Na+) and chloride ions (C l-) in NaC l can participate in ion balance adjustment and enzyme activity adjustment in microorganisms by preparing the reinforced solution, so that the growth and activity of the microorganisms can be promoted, and the calcium ions (Ca2+) in CaC l 2 can promote the stability of cell walls and cell membranes of the microorganisms, enhance the stress resistance of the microorganisms, and strengthen the structural stability and flocculation performance of the flocculant by the interaction of ions and functional groups in flocculant molecules.
Description
Technical Field
The invention relates to the technical field of flocculant processing, in particular to a preparation method of a microbial reinforced flocculant.
Background
The microbial flocculant is a kind of metabolic products produced by microorganisms or secretion thereof, is obtained by fermentation, extraction and refining of microorganisms such as bacteria and fungi, has biodegradability and safety, is a high-efficiency, nontoxic and secondary pollution-free water treatment agent, is widely applied to water treatment and wastewater treatment, can effectively remove suspended matters in water, such as suspended particles, colloid matters, microorganisms and the like, improves water quality, and improves transparency and appearance of water and improves clarity and sanitary safety of the water by reducing turbidity, suspended matters content and color in the water.
Along with the acceleration of industrialization and urban process, the water pollution problem is increasingly serious, the requirements on the water treatment technology are also higher and higher, the water treatment efficiency is influenced by the activity of the existing microbial flocculant, the water treatment cost is increased by increasing the use amount of the microbial flocculant, meanwhile, the ecological balance of the water body is possibly changed by adding a large amount of microbial flocculant, the survival and propagation of aquatic organisms are influenced, the water quality is polluted, the flocculation performance of the microbial flocculant is possibly reduced, and even the reverse effect is generated, namely suspended matters are redispersed in the water, so that the water quality is deteriorated.
Increasing the activity of microbial flocculant is a key to solve the problem of complex water pollution, so we make an improvement on the problem and put forward a preparation method of microbial enhanced flocculant.
Disclosure of Invention
The present invention has been made in view of the above-mentioned existing need to increase the activity of microbial flocculant to solve the problem of complex water pollution.
Accordingly, the present invention aims to provide a method for preparing a microbial enhanced flocculant, which aims to: the structural stability and flocculation performance of the flocculant are enhanced by preparing a microbial culture medium and adding ions to interact with functional groups in flocculant molecules.
In order to solve the technical problems, the invention provides the following technical scheme: a preparation method of a microbial enhanced flocculant comprises the following steps:
preparing a microbial culture medium: selecting glucose, xanthan gum, levoglucosides, pullulan alpha-glucan and soluble starch to form a polysaccharide culture medium;
And (3) preparing a reinforcing agent: naCl and CaCl 2 are selected to be configured as strengthening solution;
the method for screening and preparing the microbial flocculant specifically comprises the following steps:
(1) Culturing polysaccharide at 25-30 deg.c and 100-130 r/min, shaking culture for 25d, collecting culture liquid, filtering to eliminate thallus to obtain fermented liquid, freezing and centrifuging for 10-15 min to obtain supernatant to form solution A;
(2) Adding emulsifying enzyme into the strengthening solution, controlling the temperature to be 30 ℃, and rapidly oscillating for 1min to form solution B;
(3) Adding the solution B into the solution A, controlling the temperature to 25 ℃, controlling the rotation speed of a shaking table to 150r/min, adding MgCl 2 solution as a coagulant aid after shaking culture for 72h, and taking supernatant after rapid shaking for 1min to form the microbial enhanced flocculant;
(4) Taking half dose of the microbial enhanced flocculant, quick-freezing and drying, and grinding to form flocculant freeze-dried powder.
As a preferred embodiment of the method for producing a microbial-enhanced flocculant of the present invention, wherein: the molar ratio of the glucose, the xanthan gum, the levoglucosides, the pullulan and the soluble starch is 1:26 to 54:1: 80-110: 10 to 23:2 to 6.
As a preferred embodiment of the method for producing a microbial-enhanced flocculant of the present invention, wherein: the NaC L is 2mmo L/L, and CaC L 2 is 4mmo L/L.
As a preferred embodiment of the method for producing a microbial-enhanced flocculant of the present invention, wherein: the fermentation broth in the step (1) is extracted by the following steps:
1) Transferring the collected culture solution into a filter, wherein the pore diameter of the filter membrane is 0.22 mu m;
2) Pressurizing the filter using a vacuum pump;
3) After the filtration was completed, the filtrate exuded from the filter membrane was taken as a fermentation broth.
As a preferred embodiment of the method for producing a microbial-enhanced flocculant of the present invention, wherein: the step of extracting the supernatant in the step (1) is as follows:
1) Sucking out the supernatant in the centrifuge tube by using a sterile pipette;
2) Centrifuging the sediment in the centrifuge tube again, standing for sedimentation, and taking out supernatant;
3) The two supernatants were mixed.
As a preferred embodiment of the method for producing a microbial-enhanced flocculant of the present invention, wherein: the specific operation steps in the step (2) are as follows:
1) Adding glucose solution into the emulsifying enzyme, and rapidly oscillating at normal temperature for 2min;
2) Mixing the reinforcing solution of NaC l and CaCl 2, and rapidly oscillating for 1min at normal temperature;
3) After controlling the ambient temperature to 30 ℃, adding the mixed solution of the emulsifying enzyme, and rapidly oscillating for 1min.
As a preferred embodiment of the method for producing a microbial-enhanced flocculant of the present invention, wherein: the flocculant freeze-dried powder in the step (4) is used as follows:
1) Mixing the microbial enhanced flocculant in the step (3) with the flocculant freeze-dried powder in a ratio of 5:1;
2) Firstly, centrifuging for 8-10 min, and then rapidly oscillating for 1min, and then throwing.
The invention has the beneficial effects that:
1. According to the invention, naCl and CaCl 2 are common inorganic salts by preparing the strengthening solution, the use cost is low, sodium ions (Na+) and chloride ions (C l-) in NaCl can participate in ion balance regulation and enzyme activity regulation in microorganisms, the growth and activity of the microorganisms are promoted, calcium ions (Ca2+) in CaCl 2 can promote the stability of cell walls and cell membranes of the microorganisms, the stress resistance of the microorganisms is enhanced, and the structural stability and flocculation performance of the flocculant are enhanced by the interaction of ions and functional groups in flocculant molecules, so that the microbial strengthening flocculant is realized.
2. According to the invention, the glucose is used as a main carbon source to provide energy for microorganisms, the xanthan gum, the L-glycoside and the like provide different kinds of polysaccharides, the growth and metabolism of the microorganisms are facilitated, the soluble starch provides a stable carbon source, the stability of energy supply of the microorganisms in the growth process is ensured, and the synthesis speed and yield of the microbial flocculant are improved by adding and mixing the emulsifying enzyme.
3. The invention can reduce the dosage of the microbial enhanced flocculant on the premise of ensuring the flocculation effect by using the flocculant freeze-dried powder, thereby reducing the production cost, and the flocculant in the form of the freeze-dried powder has the characteristics of longer quality guarantee period and convenient transportation, and also reduces the storage and transportation cost.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Wherein:
FIG. 1 is a schematic diagram showing the results of comparative tests in examples of the method for producing a microbial-enhanced flocculant of the present invention.
Detailed Description
In order that the above-recited objects, features and advantages of the present invention will become more readily apparent, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
Example 1
Referring to fig. 1, for a first embodiment of the present invention, there is provided a method for preparing a microorganism-enhanced flocculant, comprising the steps of:
preparing a microbial culture medium: selecting glucose, xanthan gum, levoglucosides, pullulan alpha-glucan and soluble starch to form a polysaccharide culture medium;
And (3) preparing a reinforcing agent: naCl and CaCl 2 are selected to be configured as strengthening solution;
the method for screening and preparing the microbial flocculant specifically comprises the following steps:
(1) Culturing polysaccharide at 25-30 deg.c and 100-130 r/min, shaking culture for 25d, collecting culture liquid, filtering to eliminate thallus to obtain fermented liquid, freezing and centrifuging for 10-15 min to obtain supernatant to form solution A;
(2) Adding emulsifying enzyme into the strengthening solution, controlling the temperature to be 30 ℃, and rapidly oscillating for 1min to form solution B;
(3) Adding the solution B into the solution A, controlling the temperature to 25 ℃, controlling the rotation speed of a shaking table to 150r/min, adding MgCl 2 solution as a coagulant aid after shaking culture for 72h, and taking supernatant after rapid shaking for 1min to form the microbial enhanced flocculant;
(4) Taking half dose of the microbial enhanced flocculant, quick-freezing and drying, and grinding to form flocculant freeze-dried powder.
In this embodiment, experiments are performed by selecting sewage in the same environment, and setting three groups of comparison groups of no-enhancer group, no-MgCl 2 solution in the added-enhancer group and no-MgCl 2 solution in the enhancer at the same temperature, and the experimental results refer to the attached figure 1 of the specification, and the experimental results show that the addition of Na+, mg2 and Ca2 can promote flocculation activity of the microbial culture medium.
The microbial enhanced flocculant is environment-friendly, the used raw materials are natural polysaccharide and inorganic salt, no toxic or harmful substances are contained, waste generated in the preparation process is easy to treat, pollution to the environment is avoided, naCl and CaCl 2 are selected as enhanced solutions, mgCl 2 solution is added as coagulant aid, na+, mg2 and Ca2 are added to promote flocculation activity of the microbial culture medium, and the microbial enhanced flocculant is low in cost, efficient in treatment, environment-friendly and good in application prospect.
Example 2
For the second embodiment of the present invention, this embodiment is different from the first embodiment in that:
Preparing a microbial culture medium: the polysaccharide culture medium is composed of glucose, xanthan gum, L-glucoside, pullulan alpha-glucan and soluble starch, wherein the molar ratio of the glucose, the xanthan gum, the L-glucoside, the pullulan alpha-glucan and the soluble starch is 1:26 to 54:1: 80-110: 10 to 23: 2-6, providing energy for microorganisms by using glucose as a main carbon source; xanthan gum, L-glucoside and the like provide different kinds of polysaccharide, and are beneficial to the growth and metabolism of microorganisms; the soluble starch provides a stable carbon source, ensuring the stability of the energy supply of the microorganism during growth.
And (3) preparing a reinforcing agent: naC L and CaC L 2 are selected to be configured as strengthening solutions, wherein NaC L is 2mmo L/L, caC L 2 is 4mmo L/L, naC L and CaC L 2 are common inorganic salts, the price is relatively low, the sources are wide, the cost of the microbial flocculant is reduced, the growth and metabolic processes of the microorganism are positively influenced by providing different types of ions, sodium ions (Na+) and chloride ions (C L-) in NaC L can participate in ion balance regulation and enzyme activity regulation in the microorganism, the growth and activity of the microorganism are promoted, calcium ions (Ca2+) in CaC L 2 are key participants of many biochemical reactions, the stability of cell walls and cell membranes of the microorganism can be promoted, the stress resistance of the microorganism is enhanced, the ions interact with functional groups in flocculant molecules, and the structural stability and flocculation performance of the flocculant are enhanced.
Example 3
For the third embodiment of the present invention, this embodiment is different from the second embodiment in that:
the fermentation broth in the step (1) is extracted by the following steps:
1) Transferring the collected culture solution into a filter, wherein the pore diameter of the filter membrane is 0.22 mu m;
2) Pressurizing the filter using a vacuum pump;
3) After the filtration was completed, the filtrate exuded from the filter membrane was taken as a fermentation broth.
In this embodiment, by filtering instead of centrifugal mechanical separation methods, active components in the fermentation broth can be retained to the greatest extent, and compared with natural filtration, the use of a vacuum pump to pressurize the filter can accelerate the filtration process, improve the operation efficiency, and the purity of the fermentation broth extracted by the filtration method is high, so that the pure fermentation broth is helpful to reduce the interference of impurities on subsequent steps, and improve the quality and performance of the final product.
Example 4
For the third embodiment of the present invention, this embodiment is different from the second embodiment in that:
the step of extracting the supernatant in the step (1) is as follows:
1) Sucking out the supernatant in the centrifuge tube by using a sterile pipette;
2) Centrifuging the sediment in the centrifuge tube again, standing for sedimentation, and taking out supernatant;
3) The two supernatants were mixed.
In the embodiment, the microbial flocculant and other active ingredients in the supernatant can be ensured to be purer through repeated centrifugation and standing precipitation, the influence of impurities on the flocculant performance is avoided, and meanwhile, the operation is also helpful for removing insoluble substances or suspended matters in the supernatant, so that the supernatant is clearer and more transparent, the synthesis process of the microbial flocculant can be ensured to be smoother, and the activity and stability of the product are improved.
Example 5
For the third embodiment of the present invention, this embodiment is different from the second embodiment in that:
The specific operation steps in the step (2) are as follows:
1) Adding glucose solution into the emulsifying enzyme, and rapidly oscillating at normal temperature for 2min;
2) Mixing the reinforcing solution of NaC l and CaCl 2, and rapidly oscillating for 1min at normal temperature;
3) After controlling the ambient temperature to 30 ℃, adding the mixed solution of the emulsifying enzyme, and rapidly oscillating for 1min.
In this example, the dissolution and dispersion of the emulsifying enzyme can be effectively promoted by adding the emulsifying enzyme to the glucose solution and performing rapid shaking, and the glucose is used as a carbon source to supply energy to the emulsifying enzyme so as to enhance the activity thereof, and the environmental temperature is controlled to be 30 ℃ before the emulsifying enzyme mixed solution is added, which is a suitable temperature at which the emulsifying enzyme and the enhancer act. By controlling the temperature, the reaction process can be ensured to be carried out under the optimal condition, thereby improving the synthesis speed and the yield of the microbial flocculant.
Example 6
For the third embodiment of the present invention, this embodiment is different from the second embodiment in that:
the flocculant freeze-dried powder in the step (4) is used as follows:
1) Mixing the microbial enhanced flocculant in the step (3) with the flocculant freeze-dried powder in a ratio of 5:1;
2) Firstly, centrifuging for 8-10 min, and then rapidly oscillating for 1min, and then throwing.
In this embodiment, the flocculant in the form of the freeze-dried powder has higher activity and stability, can be quickly fused with the microbial strengthening flocculant, improves the overall flocculation performance, can quickly remove impurities and insoluble particles in the mixed solution through centrifugal treatment, so that the flocculant is purer, and can reduce the dosage of the microbial strengthening flocculant on the premise of ensuring the flocculation effect by using the flocculant freeze-dried powder, thereby reducing the production cost.
It should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered in the scope of the claims of the present invention.
Claims (7)
1. A preparation method of a microbial enhanced flocculant is characterized by comprising the following steps: the method comprises the following steps:
preparing a microbial culture medium: selecting glucose, xanthan gum, levoglucosides, pullulan alpha-glucan and soluble starch to form a polysaccharide culture medium;
And (3) preparing a reinforcing agent: naCl and CaCl 2 are selected to be configured as strengthening solution;
the method for screening and preparing the microbial flocculant specifically comprises the following steps:
(1) Culturing polysaccharide at 25-30 deg.c and 100-130 r/min, shaking culture for 25d, collecting culture liquid, filtering to eliminate thallus to obtain fermented liquid, freezing and centrifuging for 10-15 min to obtain supernatant to form solution A;
(2) Adding emulsifying enzyme into the strengthening solution, controlling the temperature to be 30 ℃, and rapidly oscillating for 1min to form solution B;
(3) Adding the solution B into the solution A, controlling the temperature to 25 ℃, controlling the rotation speed of a shaking table to 150r/min, adding MgCl 2 solution as a coagulant aid after shaking culture for 72h, and taking supernatant after rapid shaking for 1min to form the microbial enhanced flocculant;
(4) Taking half dose of the microbial enhanced flocculant, quick-freezing and drying, and grinding to form flocculant freeze-dried powder.
2. The method for preparing a microbial enhanced flocculant according to claim 1, wherein: the molar ratio of the glucose, the xanthan gum, the levoglucosides, the pullulan and the soluble starch is 1:26 to 54:1: 80-110: 10 to 23:2 to 6.
3. The method for preparing a microbial enhanced flocculant according to claim 2, wherein: the NaCl is 2mmol/L, and CaCl 2 is 4mmol/L.
4. A method of preparing a microbial enhanced flocculant according to claim 3, wherein: the fermentation broth in the step (1) is extracted by the following steps:
1) Transferring the collected culture solution into a filter, wherein the pore diameter of the filter membrane is 0.22 mu m;
2) Pressurizing the filter using a vacuum pump;
3) After the filtration was completed, the filtrate exuded from the filter membrane was taken as a fermentation broth.
5. The method for preparing a microbial enhanced flocculant according to claim 4, wherein: the step of extracting the supernatant in the step (1) is as follows:
1) Sucking out the supernatant in the centrifuge tube by using a sterile pipette;
2) Centrifuging the sediment in the centrifuge tube again, standing for sedimentation, and taking out supernatant;
3) The two supernatants were mixed.
6. The method for preparing a microbial enhanced flocculant according to claim 5, wherein: the specific operation steps in the step (2) are as follows:
1) Adding glucose solution into the emulsifying enzyme, and rapidly oscillating at normal temperature for 2min;
2) Mixing the reinforced solution of NaCl and CaCl 2, and rapidly oscillating for 1min at normal temperature;
3) After controlling the ambient temperature to 30 ℃, adding the mixed solution of the emulsifying enzyme, and rapidly oscillating for 1min.
7. The method for preparing a microbial enhanced flocculant according to claim 6, wherein: the flocculant freeze-dried powder in the step (4) is used as follows:
1) Mixing the microbial enhanced flocculant in the step (3) with the flocculant freeze-dried powder in a ratio of 5:1;
2) Firstly, centrifuging for 8-10 min, and then rapidly oscillating for 1min, and then throwing.
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