CN111995026A - Environment-friendly efficient composite biological flocculant and preparation method thereof - Google Patents
Environment-friendly efficient composite biological flocculant and preparation method thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
- C02F1/54—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using organic material
- C02F1/56—Macromolecular compounds
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
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- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
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Abstract
The invention discloses an environment-friendly efficient composite biological flocculant and a preparation method thereof, and relates to the technical field of flocculant preparation. The preparation method of the environment-friendly efficient composite bioflocculant comprises the following steps: adding dipotassium hydrogen phosphate into the bean curd wastewater, stirring and sterilizing to obtain a fermentation culture solution; inoculating microbial seed liquid, sealing, fermenting, centrifuging, taking supernatant, adding absolute ethyl alcohol, stirring and freezing to obtain various microbial flocculants. Dissolving sodium hydroxide and tartaric acid solid in deionized water, adding acrylamide monomer and locust bean gum aqueous solution, stirring, adjusting pH value, charging nitrogen gas, then adding initiator, standing after ultraviolet irradiation, washing, purifying, and drying to obtain the modified natural polymeric flocculant. And compounding the modified natural polymeric flocculant and the microbial flocculant mixture to obtain the environment-friendly high-efficiency composite biological flocculant. The composite biological flocculant has high flocculation performance, can reduce the dosage of the flocculant, and is safe, nontoxic and free of secondary pollution.
Description
Technical Field
The invention belongs to the technical field of flocculant preparation, and particularly relates to an environment-friendly efficient composite biological flocculant and a preparation method thereof.
Background
Water resources are important resources indispensable to the development of the human society and to the maintenance of various animal and plant life activities. Along with the rapid growth of population, the rapid development of social economy and the accelerated promotion of industrialization process, the contradiction between the industrial wastewater production and the domestic water demand produced by population life is sharp, and the crisis of water resources is obvious and urgent. Therefore, the problem of water treatment (including raw water treatment, sewage treatment, and industrial water treatment, etc.) has become more and more severe. Flocculation is one of the most common methods for removing particulate matter and organic matter from water, and a flocculant is one of the most important factors for determining flocculation effect.
At present, the most commonly used flocculating agents in the water treatment process are mainly inorganic flocculating agents and organic synthetic polymer flocculating agents, which cause certain pollution to the environment and even influence the health of human beings. The microbial flocculant is a secondary metabolite which is generated by microorganisms and can cause suspended particles, somatic cells and colloidal particles in a water body to coagulate and precipitate, mainly contains substances such as glycoprotein, polysaccharide, protein, cellulose, deoxyribonucleic acid and the like, is a high-efficiency, non-toxic and secondary-pollution-free green water treatment agent with biodegradability and safety, and has become one of hot spots of domestic and foreign researches in recent years. However, because the production cost of the microbial flocculant is high, the fermentation production process is not mature, and the molecular structure, shape, molecular mass, carried groups and the like of the microbial flocculant influence the activity of the flocculant, so that the development of the microbial flocculant is limited to a certain extent. The search for suitable composite biological flocculants is a new direction for preparing efficient, environment-friendly and safe flocculants.
Disclosure of Invention
The invention aims to provide an environment-friendly efficient composite bioflocculant and a preparation method thereof.
The technical scheme adopted by the invention for realizing the purpose is as follows:
a preparation method of an environment-friendly efficient composite biological flocculant comprises the following steps:
s1: adjusting the pH value of the bean curd wastewater, adding dipotassium hydrogen phosphate, stirring and sterilizing to obtain a fermentation culture solution;
s2: inoculating various microorganism seed solutions into the fermentation culture solution, and performing sealed fermentation culture to obtain fermentation strain solutions; centrifuging, adding absolute ethyl alcohol into the supernatant, stirring and freezing to obtain various microbial flocculants;
s3: dissolving sodium hydroxide and tartaric acid solid in deionized water, adding acrylamide monomer and locust bean gum aqueous solution, and stirring; adjusting the pH value of the reaction solution, filling nitrogen, then adding an initiator aqueous solution, sealing, irradiating by ultraviolet light, standing and washing; purifying with N, N-dimethyl amide, drying at constant temperature until the quality is unchanged, and grinding to obtain a modified natural polymeric flocculant;
s4: and (4) dissolving the mixture of the modified natural polymeric flocculant in the step S3 and the microbial flocculants in the step S2 in water, stirring, standing, performing centrifugal precipitation, and performing vacuum drying to obtain the composite biological flocculant. The composite biological flocculant is constructed by compounding a microbial flocculant mixture and a modified natural polymeric flocculant, the microbial flocculant is non-toxic, harmless and biodegradable, and the locust bean gum serving as a matrix of the modified polymeric flocculant has high safety and is often applied to the food industry. The two are compounded to ensure the safety of the composite biological flocculant, no secondary pollution is caused, and the composite biological flocculant belongs to a green environment-friendly flocculant. The locust bean gum has great amount of hydroxyl active groups in its structure, and is used as natural polymer compound through chemical modification of grafting copolymerization and other chemical modification, so as to improve the flocculation performance and dissolving performance greatly. The polyacrylamide is grafted to the molecular chain of the locust bean gum, so that the biological solubility of the locust bean gum is improved, the molecular weight is increased, the cationic degree is improved, and the flocculation effect of the composite bioflocculant is further improved; tartaric acid is an unsaturated dibasic acid, contains active double bonds and two carboxyl groups in the molecule, is a bio-based monomer with higher polymerization activity, is grafted to the molecular chain of the locust bean gum, enhances the bridging effect and the characteristic adsorption effect, and can improve the flocculation performance of the composite bioflocculant.
Preferably, the pH value of the bean curd wastewater in the step S1 is 6.8-7, the content of dipotassium hydrogen phosphate is 4-4.2 g/L, the sterilization temperature is 120-130 ℃, and the sterilization time is 20-30 min.
Preferably, the microorganism seed solution in step S2 includes: rhodococcus erythropolis, Aspergillus fumigatus, Bacillus subtilis; the inoculation amount of each microorganism is 5-8%, the fermentation temperature is 28-37 ℃, and the culture time is 60-80 h.
Preferably, the amount of the absolute ethyl alcohol added in the step S2 is 1.5-3 times of the volume of the supernatant; the centrifugal speed is 5800-7600 r/min, the centrifugal time is 11-13 min, the freezing temperature is 2-5 ℃, and the freezing time is 25-45 h.
Preferably, the molar ratio of the sodium hydroxide to the tartaric acid in the step S3 is 0.9-1: 1; the molar ratio of the acrylamide monomer to the tartaric acid is 1-1.2: 1; the mass fraction of the locust bean gum aqueous solution is 3-5%, the content of the locust bean gum is 15-18% of the total amount of the acrylamide monomer, and the initiator is azobisisobutyrimidazoline hydrochloride, the content of the initiator is 0.06-0.08% of the total amount of the acrylamide monomer.
Preferably, the pH value of the reaction solution in the step S3 is 4.0-5.0; the ultraviolet irradiation time is 110-125 min.
Preferably, in the step S3, the standing time is 2-3 h, the washing solution is acetone, and the constant-temperature drying temperature is (60 +/-2) DEG C.
Preferably, the mixture of microbial flocculants in step S4 includes: 30-45 parts by weight of a flocculating agent obtained by fermenting rhodococcus erythropolis, 10-25 parts by weight of a flocculating agent obtained by fermenting aspergillus fumigatus and 20-35 parts by weight of a flocculating agent obtained by fermenting bacillus subtilis.
Preferably, the composite bioflocculant in step S4 includes: 70-85 parts of a microbial flocculant mixture and 15-30 parts of a natural polymer modified flocculant by weight.
Preferably, in the step S4, the stirring time is 45-60 min, and the vacuum drying temperature is 55-60 ℃.
Preferably, the sodium arachidonic acid and the absolute ethyl alcohol are added into the fermentation strain liquid of the step S2, and the mixture is stirred to obtain modified microbial flocculants; wherein the volume ratio of the mass of the arachidonic acid sodium to the fermentation strain liquid is 1-1.1 g: 30 mL; stirring for 40-50 min; the volume ratio of the absolute ethyl alcohol to the fermentation strain liquid is 0.8-1: 5. One of the main components of the macromolecular polymer secreted outside cells by microorganisms is extracellular polysaccharide which has more hydrophilic groups such as hydroxyl, carboxyl and the like, and the aggregation of a microbial flocculant and target flocculate is hindered to a certain extent. After the arachidonic acid sodium salt in the mass-volume ratio range is added, the secretion of microbial extracellular polysaccharide can be promoted, and meanwhile, the arachidonic acid sodium salt can be combined with a microbial flocculant to reduce hydrophilic groups on the surface of the microbial flocculant and increase hydrophobic groups on the surface of the microbial flocculant, so that the flocculation performance of the microbial flocculant on flocculates is improved.
The invention also discloses an environment-friendly high-efficiency composite bioflocculant prepared by the preparation method.
The invention adopts the modified natural polymeric flocculant and the microbial flocculant to compound, thereby having the following beneficial effects:
the composite biological flocculant is constructed by compounding a microbial flocculant mixture and a natural polymer modified flocculant locust bean gum, the microbial flocculant is non-toxic, harmless and biodegradable, and the polymer modified flocculant locust bean gum has high safety and is often applied to the food industry. The safety of the composite biological flocculant is ensured, no secondary pollution is caused, and the composite biological flocculant is environment-friendly. Through chemical modification such as grafting/graft copolymerization, the flocculation effect and the solubility of the natural polymeric flocculant are improved, the natural polymeric flocculant is further compounded with the microbial flocculant, the bridging effect and the characteristic adsorption effect of the microbial flocculant are enhanced, and the flocculation performance of the composite biological flocculant is further improved. The microbial flocculant is modified by adding the arachidonic acid sodium, so that the secretion of extracellular polysaccharide of the microbes is promoted, meanwhile, the hydrophobic groups on the surface of the microbes can be increased, and the flocculation performance of the microbes on flocculates is further improved. The composite biological flocculant of the invention has less addition amount and convenient operation.
Therefore, the invention provides an environment-friendly high-efficiency composite biological flocculant and a preparation method thereof.
Drawings
FIG. 1 is a comparative graph showing the results of the flocculation test on the kaolin suspension in test example 1 of the present invention;
FIG. 2 is a comparison diagram of the results of tests on flocculation effect and COD removal rate of actual papermaking wastewater in test example 2 of the present invention;
FIG. 3 is a comparison diagram of the results of the flocculation effect and COD removal rate tests on municipal sewage in test example 3 of the present invention;
FIG. 4 is a comparison of the results of the measurement of the extracellular polysaccharide content of the microorganism in test example 4 of the present invention.
Detailed Description
The technical solution of the present invention is further described in detail below with reference to the following detailed description and the accompanying drawings:
example 1:
a preparation method of an environment-friendly efficient composite biological flocculant comprises the following steps:
s1: taking 50mL of bean curd wastewater, adjusting the pH to 7, adding 0.2g of dipotassium phosphate, stirring, and sterilizing at 121 ℃ for 20min to obtain a fermentation culture solution;
s2: respectively inoculating Rhodococcus erythropolis, Aspergillus fumigatus and Bacillus subtilis to the fermentation culture solution according to the inoculum size of 6%, sealing, fermenting and culturing at 31 deg.C under shaking of 140r/min in incubator for 80 h; centrifuging at 7000r/min for 10min, collecting supernatant, adding 2 times of anhydrous ethanol, stirring, and freezing at 2 deg.C for 40min to obtain microbial flocculants;
s3: weighing sodium hydroxide and tartaric acid solids with the same molar mass, dissolving in deionized water, and mixing the components according to the weight ratio of acrylamide monomer: adding acrylamide monomer into tartaric acid at a molar mass ratio of 1.2:1, adding 3% locust bean gum aqueous solution (the content of the locust bean gum is 16% of the total amount of the acrylamide monomer), and stirring to form a uniform solution; the concentration of the catalyst is 0.1 mol.L-1Adjusting pH of the reaction solution to 5 with hydrochloric acid solution and sodium hydroxide solution, adding 0.06% azobisisobutyrimidazoline hydrochloride, rapidly sealing, placing in ultraviolet device (power of 500W, dominant wavelength of 365nm, light intensity of 1200 μ W cm)-2) Irradiating for 120min, standing and curing for 2h, extracting a product in a reaction bottle by using acetone, and washing for multiple times to obtain a crude product; then the product is subjected to Soxhlet purification by using N, N-dimethyl amide to remove homopolymerizationPlacing the mixture and unreacted monomers in an oven, drying at a constant temperature of 60 ℃ until the mass of the mixture is unchanged, and grinding to obtain a modified natural polymeric flocculant;
s4: and (2) dissolving 20 parts by weight of the modified natural polymeric flocculant in the step S3 and 80 parts by weight of the microbial flocculant mixture in the step S2 (wherein 35 parts by weight of the flocculant obtained by fermenting rhodococcus erythropolis, 20 parts by weight of the flocculant obtained by fermenting aspergillus fumigatus and 25 parts by weight of the flocculant obtained by fermenting bacillus subtilis) in water, stirring for 50min at 130rpm, standing, centrifuging, collecting precipitate, and drying in vacuum at 60 ℃ to obtain the composite biological flocculant.
Example 2:
a preparation method of an environment-friendly efficient composite biological flocculant comprises the following steps:
s1: taking 50mL of bean curd wastewater, adjusting the pH to 7, adding 0.2g of dipotassium phosphate, stirring, and sterilizing at 121 ℃ for 20min to obtain a fermentation culture solution;
s2: respectively inoculating Rhodococcus erythropolis, Aspergillus fumigatus and Bacillus subtilis to the fermentation culture solution according to the inoculum size of 6%, sealing, fermenting and culturing at 31 deg.C under shaking of 140r/min in incubator for 80h to obtain fermentation strain solution; then, according to the mass-volume ratio of the arachidonic acid sodium to the fermentation strain liquid of 1 g: adding arachidonic acid sodium in an amount of 30mL, adding absolute ethyl alcohol according to the volume ratio of the absolute ethyl alcohol to the fermentation strain liquid of 1:5, and stirring for 45 min; centrifuging at 7000r/min for 10min, collecting supernatant, adding 2 times of anhydrous ethanol, stirring, and freezing at 2 deg.C for 40min to obtain modified microbial flocculants;
s3: weighing sodium hydroxide and tartaric acid solids with the same molar mass, dissolving in deionized water, and mixing the components according to the weight ratio of acrylamide monomer: adding acrylamide monomer into tartaric acid at a molar mass ratio of 1.2:1, adding 3% locust bean gum aqueous solution (the content of the locust bean gum is 16% of the total amount of the acrylamide monomer), and stirring to form a uniform solution; adjusting the pH value of the reaction solution to 5 by using hydrochloric acid solution with the concentration of 0.1 mol.L < -1 > and sodium hydroxide solution, then adding 0.06 percent of azobisisobutyrimidazoline hydrochloride, quickly sealing, placing in an ultraviolet device (the power is 500W, the dominant wavelength is 365nm, and the light intensity is 1200 mu W.cm < -2 >) for irradiating for 120min, standing and curing for 2h, extracting the product in a reaction bottle by using acetone, and washing for multiple times to obtain a crude product; performing Soxhlet purification on the product by using N, N-dimethyl amide to remove homopolymers and unreacted monomers in the product, then placing the product in an oven for drying at a constant temperature of 60 ℃ until the quality is unchanged, and grinding the product to obtain a modified natural polymeric flocculant;
s4: and (2) dissolving 20 parts by weight of the modified natural polymeric flocculant obtained in the step S3 and 80 parts by weight of the microbial flocculant mixture modified in the step S2 (wherein 35 parts by weight of the flocculant obtained by fermenting rhodococcus erythropolis, 20 parts by weight of the flocculant obtained by fermenting aspergillus fumigatus and 25 parts by weight of the flocculant obtained by fermenting bacillus subtilis) in water, stirring for 50min at 130rpm, standing, centrifuging, collecting precipitate, and drying in vacuum at 60 ℃ to obtain the composite biological flocculant.
Comparative example 1:
a preparation method of a composite biological flocculant comprises the following steps:
s1: taking 50mL of bean curd wastewater, adjusting the pH to 7, adding 0.2g of dipotassium phosphate, stirring, and sterilizing at 121 ℃ for 20min to obtain a fermentation culture solution;
s2: respectively inoculating Rhodococcus erythropolis, Aspergillus fumigatus and Bacillus subtilis to the fermentation culture solution according to the inoculum size of 6%, sealing, fermenting and culturing at 31 deg.C under shaking of 140r/min in incubator for 80 h; centrifuging at 7000r/min for 10min, collecting supernatant, adding 2 times of anhydrous ethanol, stirring, and freezing at 2 deg.C for 40min to obtain microbial flocculants; taking 35 parts by weight of a flocculant obtained by fermenting rhodococcus erythropolis, 20 parts by weight of a flocculant obtained by fermenting aspergillus fumigatus and 25 parts by weight of a flocculant obtained by fermenting bacillus subtilis, and mixing the two parts in water to obtain the microbial flocculant blend.
Comparative example 2:
a preparation method of a composite biological flocculant comprises the following steps:
s1: taking 50mL of bean curd wastewater, adjusting the pH to 7, adding 0.2g of dipotassium phosphate, stirring, and sterilizing at 121 ℃ for 20min to obtain a fermentation culture solution;
s2: respectively inoculating Rhodococcus erythropolis, Aspergillus fumigatus and Bacillus subtilis to the fermentation culture solution according to the inoculum size of 6%, sealing, fermenting and culturing at 31 deg.C under shaking of 140r/min in incubator for 80 h; centrifuging at 7000r/min for 10min, collecting supernatant, adding 2 times of anhydrous ethanol, stirring, and freezing at 2 deg.C for 40min to obtain microbial flocculants;
s3: according to the weight parts, 20 parts of locust bean gum base natural polymeric flocculant and 80 parts of each microbial flocculant mixture obtained in the step S2 (wherein, 35 parts of flocculant obtained by fermenting rhodococcus erythropolis, 20 parts of flocculant obtained by fermenting aspergillus fumigatus and 25 parts of flocculant obtained by fermenting bacillus subtilis) are dissolved in water together, stirred for 50min at 130rpm, kept stand, centrifuged to collect precipitate, and dried in vacuum at 60 ℃ to obtain the composite biological flocculant.
Comparative example 3:
a preparation method of a composite biological flocculant comprises the following steps:
s1: taking 50mL of bean curd wastewater, adjusting the pH to 7, adding 0.2g of dipotassium phosphate, stirring, and sterilizing at 121 ℃ for 20min to obtain a fermentation culture solution;
s2: respectively inoculating Rhodococcus erythropolis, Aspergillus fumigatus and Bacillus subtilis to the fermentation culture solution according to the inoculum size of 6%, sealing, fermenting and culturing at 31 deg.C under shaking of 140r/min in incubator for 80 h; centrifuging at 7000r/min for 10min, collecting supernatant, adding 2 times of anhydrous ethanol, stirring, and freezing at 2 deg.C for 40min to obtain microbial flocculants;
s3: adding acrylamide monomer into deionized water, adding 3 wt% of locust bean gum aqueous solution (the content of the locust bean gum is 16% of the total amount of the acrylamide monomer), and stirring to form a uniform solution; adjusting the pH value of the reaction solution to 5 by using hydrochloric acid solution with the concentration of 0.1 mol.L < -1 > and sodium hydroxide solution, then adding 0.06 percent of azobisisobutyrimidazoline hydrochloride, quickly sealing, placing in an ultraviolet device (the power is 500W, the dominant wavelength is 365nm, and the light intensity is 1200 mu W.cm < -2 >) for irradiating for 120min, standing and curing for 2h, extracting the product in a reaction bottle by using acetone, and washing for multiple times to obtain a crude product; performing Soxhlet purification on the product by using N, N-dimethyl amide to remove homopolymers and unreacted monomers in the product, then placing the product in an oven for drying at a constant temperature of 60 ℃ until the quality is unchanged, and grinding the product to obtain a modified natural polymeric flocculant;
s4: and (2) dissolving 20 parts by weight of the modified natural polymeric flocculant in the step S3 and 80 parts by weight of the microbial flocculant mixture in the step S2 (wherein 35 parts by weight of the flocculant obtained by fermenting rhodococcus erythropolis, 20 parts by weight of the flocculant obtained by fermenting aspergillus fumigatus and 25 parts by weight of the flocculant obtained by fermenting bacillus subtilis) in water, stirring for 50min at 130rpm, standing, centrifuging, collecting precipitate, and drying in vacuum at 60 ℃ to obtain the composite biological flocculant.
Comparative example 4:
a preparation method of a composite biological flocculant comprises the following steps:
s1: taking 50mL of bean curd wastewater, adjusting the pH to 7, adding 0.2g of dipotassium phosphate, stirring, and sterilizing at 121 ℃ for 20min to obtain a fermentation culture solution;
s2: respectively inoculating Rhodococcus erythropolis, Aspergillus fumigatus and Bacillus subtilis to the fermentation culture solution according to the inoculum size of 6%, sealing, fermenting and culturing at 31 deg.C under shaking of 140r/min in incubator for 80 h; centrifuging at 7000r/min for 10min, collecting supernatant, adding 2 times of anhydrous ethanol, stirring, and freezing at 2 deg.C for 40min to obtain microbial flocculants;
s3: weighing sodium hydroxide and tartaric acid solid with the same molar mass, dissolving in deionized water, adding 3% locust bean gum aqueous solution (the content of locust bean gum is 16% of total amount of tartaric acid), and stirring to form uniform solution; adjusting the pH value of the reaction solution to 5 by using hydrochloric acid solution with the concentration of 0.1 mol.L < -1 > and sodium hydroxide solution, then adding 0.06 percent of azobisisobutyrimidazoline hydrochloride, quickly sealing, placing in an ultraviolet device (the power is 500W, the dominant wavelength is 365nm, and the light intensity is 1200 mu W.cm < -2 >) for irradiating for 120min, standing and curing for 2h, extracting the product in a reaction bottle by using acetone, and washing for multiple times to obtain a crude product; performing Soxhlet purification on the product by using N, N-dimethyl amide to remove homopolymers and unreacted monomers in the product, then placing the product in an oven for drying at a constant temperature of 60 ℃ until the quality is unchanged, and grinding the product to obtain a modified natural polymeric flocculant;
s4: and (2) dissolving 20 parts by weight of the modified natural polymeric flocculant in the step S3 and 80 parts by weight of the microbial flocculant mixture in the step S2 (wherein 35 parts by weight of the flocculant obtained by fermenting rhodococcus erythropolis, 20 parts by weight of the flocculant obtained by fermenting aspergillus fumigatus and 25 parts by weight of the flocculant obtained by fermenting bacillus subtilis) in water, stirring for 50min at 130rpm, standing, centrifuging, collecting precipitate, and drying in vacuum at 60 ℃ to obtain the composite biological flocculant.
Test example 1:
testing of flocculation Effect on Kaolin suspension
1.2g of kaolin (d 44 μm 325 mesh) was accurately weighed into 300mL of distilled water to prepare a 4g/L kaolin suspension, and 3mL of 1% CaCl was added2The solution was stirred and dispersed for 15min at 700 rpm. Then adding 0.3mg of composite biological flocculant, quickly stirring for 30s at the rotating speed of 400rpm, slowly stirring for 5min at the rotating speed of 100rpm, standing for 10min, taking the supernatant, measuring the absorbance at the wavelength of 550nm, and taking the absorbance of a blank group without the composite biological flocculant as a control. The flocculation rate of the kaolin suspension is calculated according to the following formula:
Y＝(A1-A2)/A1×100％
wherein, the flocculation rate of the Y-kaolin suspension is expressed in unit; a. the1-absorbance of the blank supernatant; a. the2-assay group supernatant absorbance.
The above tests were carried out for example 1, example 2, comparative example 1, comparative example 2, comparative example 3, comparative example 4, and the results of the experimental tests are shown in fig. 1. As can be seen from the figure, the flocculation performance of the composite biological flocculants prepared in the examples 1 and 2 is obviously better than that of the comparative example, and the flocculation effect can reach 99 percent at most. The flocculation effect of the composite flocculant is improved by the composite construction of the natural polymer modified flocculant and the microbial flocculant, and the flocculation effect of the composite flocculant can be further improved by modifying the structure of the microbial flocculant. The test result shows that the environment-friendly efficient composite biological flocculant prepared by the invention has high activity, excellent flocculation effect and no secondary pollution.
Test example 2:
flocculation effect on actual papermaking wastewater (main material content is shown in table 1):
TABLE 1 papermaking wastewater
Adjusting the pH value of 300mL of actual papermaking wastewater to 7.0, adding 0.3mg of composite bioflocculant, quickly stirring for 30s at the rotation speed of 400rpm, slowly stirring for 5min at the rotation speed of 100rpm, standing for 10min, measuring the absorbance of supernatant at the wavelength of 550nm, and calculating the flocculation rate; meanwhile, measuring COD by using a potassium dichromate-ferrous ammonium sulfate titration method, and calculating the removal rate of the COD.
The above tests were carried out for example 1, example 2, comparative example 1, comparative example 2, comparative example 3, comparative example 4, and the results of the experimental tests are shown in fig. 2. As can be seen from the figure, the flocculation effect and the COD removal rate of the composite biological flocculant prepared in the example 1 and the example 2 on the actual papermaking wastewater are higher than those of the comparative example. Experimental structures show that the flocculation effect of the environment-friendly efficient composite bioflocculant on actual papermaking wastewater is over 95%, and the removal rate of COD is over 85%.
Test example 3:
flocculation effect on municipal sewage (main material content shown in table 2):
TABLE 2 municipal sewage
Adjusting the pH value of 300mL of urban sewage to 7.0, adding 0.3mg of composite bioflocculant, quickly stirring for 30s at the rotation speed of 400rpm, slowly stirring for 5min at the rotation speed of 100rpm, standing for 10min, measuring the absorbance of supernatant at the wavelength of 550nm, and calculating the flocculation rate; meanwhile, measuring COD by using a potassium dichromate-ferrous ammonium sulfate titration method, and calculating the removal rate of the COD.
The above tests were carried out for example 1, example 2, comparative example 1, comparative example 2, comparative example 3, comparative example 4, and the results of the experimental tests are shown in fig. 3. As can be seen from the figure, the flocculation effect and the COD removal rate of the composite biological flocculants prepared in the examples 1 and 2 on the municipal sewage are higher than those of the comparative examples. The microbial flocculant compound has a flocculation effect on municipal sewage of more than 96 percent and a COD removal rate of more than 85 percent.
Test example 4:
and (3) detecting the extracellular polysaccharide of the microorganism:
and detecting by adopting a phenol-sulfuric acid method. The fermentation strain liquid obtained in the step S2 in the example 1 and the fermentation strain liquid obtained in the step S2 in the example 2 after the arachidonic acid sodium salt is added are respectively taken and centrifuged for 15min at the rotating speed of 1500r/min, 0.2mL of supernatant is absorbed into a 1.5mL finger-shaped tube, 0.8mL of absolute ethyl alcohol is added into the finger-shaped tube, the finger-shaped tube is placed in a refrigerator with the temperature of-4 ℃ for standing overnight after being shaken evenly, and then the finger-shaped tube is taken out and centrifuged for 15min at the speed of 3000 r/min. Removing supernatant, dissolving precipitate with 1mL distilled water, adding 0.5mL redissolved solution into 15mL test tube (provided with a blank tube, and replacing redissolved solution with distilled water), adding 0.5mL distilled water, 1mL 5% phenol and 5mL concentrated sulfuric acid, mixing, cooling to room temperature, and measuring absorbance (OD)490nm). And calculating the extracellular polysaccharide content of the fermentation liquid according to a glucose concentration-absorbance standard curve equation and the dilution condition which are measured in parallel.
The results of the experiment are shown in FIG. 4. As can be seen from the figure, the extracellular polysaccharide content of the fermentation broth after the arachidonic acid sodium is added in the example 2 is higher than that of the example 1, which shows that the arachidonic acid sodium can promote the microorganisms to secrete the extracellular polysaccharide, and further improves the flocculation performance of the microbial flocculant.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (10)
1. A preparation method of an environment-friendly efficient composite biological flocculant comprises the following steps:
s1: adjusting the pH value of the bean curd wastewater, adding dipotassium hydrogen phosphate, stirring and sterilizing to obtain a fermentation culture solution;
s2: inoculating various microorganism seed solutions into the fermentation culture solution, and performing sealed fermentation culture to obtain fermentation strain solutions; centrifuging, adding absolute ethyl alcohol into the supernatant, stirring and freezing to obtain various microbial flocculants;
s3: dissolving sodium hydroxide and tartaric acid solid in deionized water, adding acrylamide monomer and locust bean gum aqueous solution, and stirring; adjusting the pH value of the reaction solution, filling nitrogen, then adding an initiator aqueous solution, sealing, irradiating by ultraviolet light, standing and washing; purifying with N, N-dimethyl amide, drying at constant temperature until the quality is unchanged, and grinding to obtain a modified natural polymeric flocculant;
s4: and (4) dissolving the mixture of the modified natural polymeric flocculant in the step S3 and the microbial flocculant in the step S2 in water, stirring, standing, performing centrifugal precipitation, and performing vacuum drying to obtain the composite biological flocculant.
2. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: in the step S1, the pH value of the bean curd waste water is 6.8-7, the content of dipotassium hydrogen phosphate is 4-4.2 g/L, and the sterilization temperature is 120-130 ℃.
3. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: in the step S2, the microbial seed solutions are all in logarithmic growth phase, including: rhodococcus erythropolis, Aspergillus fumigatus, Bacillus subtilis; the inoculation amount of each microorganism is 5-8%, the fermentation temperature is 28-37 ℃, the rotating speed is 140-150 r/min, and the culture time is 60-80 h.
4. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: and in the step S2, the amount of the absolute ethyl alcohol added is 1.5-3 times of the volume of the supernatant.
5. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: the molar mass ratio of the sodium hydroxide to the tartaric acid in the step S3 is 0.9-1: 1; the molar mass ratio of the acrylamide monomer to the tartaric acid is 1-1.2: 1; the mass fraction of the locust bean gum aqueous solution is 3-5%, the content of the locust bean gum is 15-18% of the total amount of the acrylamide monomer, and the initiator is azobisisobutyrimidazoline hydrochloride, the content of the initiator is 0.06-0.08% of the total amount of the acrylamide monomer.
6. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: the pH value of the reaction solution is 4.0-5.0; the ultraviolet irradiation time is 110-125 min.
7. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: the mixture of microbial flocculants in step S4 includes: 30-45 parts by weight of a flocculating agent obtained by fermenting rhodococcus erythropolis, 10-25 parts by weight of a flocculating agent obtained by fermenting aspergillus fumigatus and 20-35 parts by weight of a flocculating agent obtained by fermenting bacillus subtilis.
8. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: the composite biological flocculant in the step S4 comprises the following steps: 70-85 parts of a microbial flocculant mixture and 15-30 parts of a modified natural polymeric flocculant by weight.
9. The preparation method of the environment-friendly efficient composite bioflocculant according to claim 1, wherein the preparation method comprises the following steps: in the step S4, the stirring time is 45-60 min, and the vacuum drying temperature is 55-60 ℃.
10. The environment-friendly efficient composite biological flocculant prepared by the preparation method of claim 1.
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