CN107686172A - A kind of preparation method of compound adsorbing metal ions flocculant - Google Patents

A kind of preparation method of compound adsorbing metal ions flocculant Download PDF

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CN107686172A
CN107686172A CN201710977173.2A CN201710977173A CN107686172A CN 107686172 A CN107686172 A CN 107686172A CN 201710977173 A CN201710977173 A CN 201710977173A CN 107686172 A CN107686172 A CN 107686172A
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张芸
李璐
刘沁
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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
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    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
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    • C12P21/00—Preparation of peptides or proteins
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
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Abstract

The invention discloses a kind of preparation method of compound adsorbing metal ions flocculant, belong to flocculant technical field.The present invention separates the producing strains of four plants of microbial flocculants of flocculation activity highest from river sludge, mixed fermentation obtains microbial flocculant, generate the high-molecular compounds such as glycoprotein, mucopolysaccharide, protein, cellulose and nucleic acid, being dissolved in can make after water fine particles in water and natural micelle be condensed into bulk floccule, so as to be removed in water, they have efficient flocculating effect, improve the utilization rate and activity of flocculant.Using alpha Ketoglutarate modification of chitosan, the chitosan derivatives with compared with high-adsorption-capacity, introduction-NH have been synthesized2With outside carbonyl reaction, introducing carboxyl, stable five-membered ring chelate can be formed with metal ion, can effectively adsorb the heavy metal ion in waste water.The problem of flocculant utilization rate that the present invention solves processing sewage at present is low, and adsorptivity is poor, and activity is low.

Description

A kind of preparation method of compound adsorbing metal ions flocculant
Technical field
The invention belongs to flocculant technical field, and in particular to a kind of preparation side of compound adsorbing metal ions flocculant Method.
Background technology
With the development and frequently mankind's activity of modern industry, the heavy metal pollution in ecological environment is on the rise, closely The taking place frequently for event such as blood lead, cadmium rice again shows that heavy metal contaminants can be straight by media such as air, water body or soil over year Connect or enter human body indirectly, make human health by unprecedented threat, therefore actively develop the pollution behavior of heavy metal Research and its preventing and controlling have become the emphasis of current Chinese government's work.
The direct discharge of heavy metal containing sewage can not only cause seriously to pollute to environment, and the health for also directly influencing the mankind is raw It is living.Wherein, heavy metal cadmium has the characteristics of high toxicity, widely distributed, and people and growing for animals and plants are endangered with serious Evil.Cadmium wastewater is essentially from Electroplate Factory, dye industry, mining and metallurgy process, battery production, process hides, film manufacture and automobile making Deng department.Therefore the highest attention that cadmium pollution is environmentally protected researcher how is removed.The side of traditional processing cadmium wastewater Method mainly has ion-exchange, chemical precipitation method, membrane separation process and active carbon adsorption etc., but these method generally existings are handled Effect is bad, spends the shortcomings of high.Bioremediation technology is the processing heavy metal polluted waste water that environmental area developed recently gets up New technology.
Flocculant is mainly with just（It is negative）Electrical group is neutralized in some water with negative（Just）Electrically it is difficult to what is separated Some particles are particle, reduce its potential, are at unstable state, and cause these particles using its aggregation property Concentrate, and separated by physics or chemical method.The medicament for generally reaching this purpose and using, referred to as flocculates Agent.The general flocculant of existing in the market includes aluminium polychloride, polyaluminium sulfate, poly-ferric chloride, poly-silicic acid iron sulfate, polymerization Aluminium chloride ferrum, cationic starch, polyacrylamide etc., in actual use there is it is certain the problem of, in inorganic agent Some flocculating properties of flocculant are very poor not to possess flocculation even, and directly using mycelium, its adsorption capacity is at maximum up to 20mg/g, adsorption capacity is low, so as to cause not reaching preferable adsorption effect, therefore, invents a kind of good wadding of flocculating effect Solidifying agent has the very big market demand.
The content of the invention
The technical problems to be solved by the invention：Low for the flocculant utilization rate of processing sewage at present, adsorptivity is poor, living A kind of the problem of property is low, it is proposed that preparation method of compound adsorbing metal ions flocculant.
In order to solve the above technical problems, the present invention is using technical scheme as described below：
A kind of compound adsorbing metal ions flocculant preparation method, it is characterised in that the preparation method comprises the following steps：
（1）Take the sludge in river to add in deionized water, be placed on magnetic stirring apparatus and stir, stand, centrifugation, collect supernatant Liquid, take quality equal supernatant four parts are seeded to No.1 bacterial liquid culture medium, No. two mould fluid nutrient mediums, three respectively Cultivated in number actinomyces fluid nutrient medium, No. four fungi liquid culture mediums, by same inoculum concentration, same condition of culture, weight Multiple enrichment culture 2 times, obtains No.1, No. two, No. three, No. four enrichment culture liquid, with the physiological saline that mass fraction is 0.9% by one Number, No. two, No. three, No. four enrichment culture liquid be diluted to 10-6 dilution factors respectively, No.1, No. two, No. three, No. four richnesses after must diluting Collect nutrient solution, then four kinds of enrichment culture liquid are distinguished into streak inoculation and consolidated in corresponding No.1 bacterium solid culture medium, No. two moulds Cultivated in body culture medium, No. three actinomyces solid mediums, No. four fungus solids culture mediums, the bacterial strain that is purified, mould Bacterial strain, actinomycetes strain, fungal bacterial strain, by bacterial screening, determine XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 as microorganism The producing strains of flocculant；
（2）XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 are seeded to No.1 bacterial liquid culture medium, No. two mould liquid trainings respectively Support base, No. three actinomyces fluid nutrient mediums, culture cultivated in No. four fungi liquid culture mediums, obtain XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solutions, XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution are mixed, mixed bacteria liquid is obtained, mixed bacteria liquid is seeded to Being fermented in fermentation culture, take fermentate, centrifuged, take supernatant, acetone is added in supernatant, constant temperature preserves 24h, takes precipitation, Precipitation is washed with ether 2 ~ 3 times, vacuum drying, obtains microbial flocculant crude extract；
（3）By chitosan and distilled water mixed dissolution, chitosan solution is obtained, α-ketoglutaric acid, room are added into chitosan solution Be stirred continuously under temperature, obtain transparent viscosity solution, with mass fraction be 0.1 mol/L sodium hydroxide solution adjust pH to 4 ~ 5, stirring reaction, sodium borohydride solution is added, adjust pH to 6 ~ 7 with the hydrochloric acid solution that mass fraction is 0.2%, reaction, must react Mixed solution afterwards, mixed solution after reaction is poured into mass fraction to be stirred in 95% ethanol, is separated out, is filtered, take precipitation, will be heavy Shallow lake is washed 3 ~ 4 times with absolute ethyl alcohol, absolute ether successively, is placed in apparatus,Soxhlet's and is used ethanol continuous extraction, is dried, is obtained α -one Glutaric acid modification of chitosan powder；
（4）Microbial flocculant crude extract is well mixed with α-ketoglutaric acid modification of chitosan powder, stood, is crushed, mistake Sieve, sieving particle is collected, obtains compound adsorbing metal ions flocculant.
The step（1）Middle sludge and the mass ratio of deionized water are 1:5, four portions of supernatants of equal mass with it is corresponding No.1 bacterial liquid culture medium, No. two mould fluid nutrient mediums, No. three actinomyces fluid nutrient mediums, No. four fungi liquid culture mediums Mass ratio be 2:9.
The step（1）The formula of middle No.1 bacterium solid culture medium is according to the mass fraction, to take 3 ~ 5 parts of beef extract, egg White 6 ~ 10 parts of peptone, 15 ~ 20 parts of agar, 2 ~ 3 parts of NaCl, 1000 parts, pH7.2 ± 0.2,121 DEG C sterilizing 20min of water；No. two moulds The formula of solid medium is according to the mass fraction, to take 20 ~ 25 parts of sucrose, NaNO31 ~ 2 part, K2HPO40.5 ~ 0.8 part, MgSO40.2 ~ 0.3 part, FeSO40.01 ~ 0.03 part, 13 ~ 15 parts of agar, 1000 parts, pH7.0 ± 0.2,115 DEG C sterilizings of water 20min；The formula of No. three actinomyces solid mediums is according to the mass fraction, to take 18 ~ 20 parts of soluble starch, KNO30.6~1 Part, K2HPO40.3 ~ 0.6 part, MgSO4·7H2O0.2 ~ 0.5 part, FeSO4·7H2O0.02 ~ 0.03 part, NaCl0.4 ~ 0.7 part, fine jade 15 ~ 20 parts of fat, 1000 parts, pH7.2 ± 0.2,121 DEG C sterilizing 20min of water；The formula of No. four fungus solids culture mediums is by quality Number meter, 180 ~ 200 parts of potato is taken, 20 ~ 22 parts of sucrose, 18 ~ 20 parts of agar, 1000 parts of water, pH is naturally, 121 DEG C of sterilizings 20min。
The step（2）The formula of middle fermentation culture takes 15 ~ 18 parts of glucose for according to the mass fraction, and urea 0.4 ~ 0.6 part, 0.2 ~ 0.5 part of yeast extract, K2HPO43 ~ 5 parts, KH2PO41 ~ 2 part, NaCl0.08 ~ 0.12 part, MgSO4·7H2O0.1~ 0.3 part, (NH4)2SO40.2 ~ 0.4 part, 1000 parts, pH7.4 ± 0.2,115 DEG C sterilizing 20min of water；One, two, three, No. four relative Answering the formula of fluid nutrient medium, other compositions are identical, and corresponding fluid nutrient medium can be made to be not added with agar.
The step（2）Middle XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution in mass ratio 2:1:2:2 mixing, supernatant The mass ratio of liquid and acetone is 2:3.
The step（3）The mass ratio of middle chitosan and distilled water is 1:10, the matter of chitosan solution and α-ketoglutaric acid Amount is than being 1:2, the mol ratio of sodium borohydride and α-ketoglutaric acid is 1:8.
The step（4）The mass ratio of middle microbial flocculant crude extract and α-ketoglutaric acid modification of chitosan powder is 5:3.
Compared with other method, advantageous effects are the present invention：
（1）The present invention separates the producing strains of four plants of microbial flocculants of flocculation activity highest from river sludge, mixes Close fermentation and obtain microbial flocculant, generate the high-molecular compounds such as glycoprotein, mucopolysaccharide, protein, cellulose and nucleic acid, Being dissolved in can make after water fine particles in water and natural micelle be condensed into bulk floccule, and so as to be removed in water, they have height Flocculation is imitated, improves the utilization rate and activity of flocculant.
（2）The present invention uses α-ketoglutaric acid modification of chitosan, has synthesized the chitosan derivative with compared with high-adsorption-capacity Thing, introduction-NH2With outside carbonyl reaction, introducing carboxyl, stable five-membered ring chelate can be formed with metal ion, can be effective Adsorb the heavy metal ion in waste water.
Embodiment
Flocculant extracts raw material：The sludge in river is taken, is placed at 4 DEG C and preserves.
No.1 bacterium solid culture medium：According to the mass fraction, 3 ~ 5 parts of beef extract, 6 ~ 10 parts of peptone, agar 15 ~ 20 are taken Part, 2 ~ 3 parts of NaCl, 1000 parts, pH7.2 ± 0.2,121 DEG C sterilizing 20min of water.
No. two mould solid mediums：According to the mass fraction, 20 ~ 25 parts of sucrose, NaNO are taken31 ~ 2 part, K2HPO40.5~0.8 Part, MgSO40.2 ~ 0.3 part, FeSO40.01 ~ 0.03 part, 13 ~ 15 parts of agar, 1000 parts, pH7.0 ± 0.2,115 DEG C sterilizings of water 20min。
No. three actinomyces solid mediums：According to the mass fraction, 18 ~ 20 parts of soluble starch, KNO are taken30.6 ~ 1 part, K2HPO40.3 ~ 0.6 part, MgSO4·7H2O0.2 ~ 0.5 part, FeSO4·7H2O0.02 ~ 0.03 part, NaCl0.4 ~ 0.7 part, agar 15 ~ 20 parts, 1000 parts, pH7.2 ± 0.2,121 DEG C sterilizing 20min of water.
No. four fungus solids culture mediums：According to the mass fraction, 180 ~ 200 parts of potato is taken, 20 ~ 22 parts of sucrose, agar 18 ~ 20 parts, 1000 parts of water, pH is naturally, 121 DEG C of sterilizing 20min.
One, two, three, No. four corresponding fluid nutrient mediums：Agar is not added with, other compositions are identical, and corresponding liquid can be made Culture medium.
Fermentation culture：According to the mass fraction, 15 ~ 18 parts of glucose, 0.4 ~ 0.6 part of urea, yeast extract 0.2 ~ 0.5 are taken Part, K2HPO43 ~ 5 parts, KH2PO41 ~ 2 part, NaCl0.08 ~ 0.12 part, MgSO4·7H2O0.1 ~ 0.3 part, (NH4)2SO40.2~0.4 Part, 1000 parts, pH7.4 ± 0.2,115 DEG C sterilizing 20min of water.
The screening of bacterium for producing flocculant of microbe：By purification of bacterial bacterial strain, fungal strain, actinomycetes strain, fungal bacterial strain It is seeded to respectively in fermentation culture（One plant of inoculation is into a nutrient solution）, the quality of every kind of bacterial strain and fermented nutritive liquid Than for 1:50,25 ~ 30 DEG C of shaking table cultures 2 ~ 3 days, centrifugation, take supernatant, obtain strain fermentating liquid by kaolin with water by matter Measure ratio 1:150 are mixed and made into aqueous suspension ofkaolin, regulated value 8 ~ 9, aqueous suspension ofkaolin, the CaCl that mass fraction is 1%2Solution, Strain fermentating liquid in mass ratio 25:1:1 is well mixed, stirs 5min, mixed liquor is poured into colorimetric cylinder, static 20min, taken Liquid at 2cm under clear liquid, its absorbance is determined under 550nm wavelength with spectrophotometer, so as to determine the flocculation of every kind of strain Rate, determine XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest.
A kind of preparation method of compound adsorbing metal ions flocculant, comprises the following steps：
（1）Take sludge in mass ratio 1:5 add in the beaker equipped with deionized water, are placed on magnetic stirring apparatus and stir 10min, quiet 1 ~ 2h is put, is centrifuged, supernatant is collected, supernatant is taken into equal four parts in mass ratio 2 of quality:9 are seeded to No.1 bacterium respectively In fluid nutrient medium, No. two mould fluid nutrient mediums, No. three actinomyces fluid nutrient mediums, No. four fungi liquid culture mediums, 25 ~ 33 DEG C culture 3 ~ 5 days, by same inoculum concentration, same condition of culture, repeats enrichment culture 2 times, obtain No.1, No. two, No. three, four Number enrichment culture liquid, with the physiological saline that mass fraction is 0.9% by No.1, No. two, No. three, No. four enrichment culture liquid difference it is dilute Release to 10-6Dilution factor, No.1, No. two, No. three, No. four enrichment culture liquid after must diluting, streak inoculation is in corresponding No.1 respectively In bacterium solid culture medium, No. two mould solid mediums, No. three actinomyces solid mediums, No. four fungus solids culture mediums, 25 ~ 33 DEG C are cultivated 3 ~ 5 days, the bacterium bacterial strain purified, fungal strain, actinomycetes strain, fungal bacterial strain, are sieved by strain Choosing, determines XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest, is the producing strains of microbial flocculant；
（2）XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 are seeded to No.1 bacterial liquid culture medium, No. two mould liquid trainings respectively Support base, No. three actinomyces fluid nutrient mediums, cultivate in No. four fungi liquid culture mediums, 25 ~ 33 DEG C are cultivated 3 ~ 5 days, obtain XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution, by XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution in mass ratio 2:1:2:2 is mixed Close, obtain mixed bacteria liquid, mixed bacteria liquid is seeded in fermentation culture, 25 ~ 30 DEG C ferment 3 ~ 5 days, take fermentate, centrifuge, take Supernatant, in mass ratio 2 in supernatant:3 add acetone, and 4 DEG C of constant temperature preserve 24h, take precipitation, and precipitation is washed 2 ~ 3 times with ether, Vacuum drying, obtains microbial flocculant crude extract；
（3）By chitosan and distilled water in mass ratio 1:10 mixed dissolutions, obtain chitosan solution, into chitosan solution add α- The mass ratio of ketoglutaric acid, chitosan solution and α-ketoglutaric acid is 1:2, it is stirred continuously, obtains transparent sticky molten at room temperature Liquid, pH to 4 ~ 5 is adjusted with the sodium hydroxide solution that mass fraction is 0.1 mol/L, stirring reaction 4h, is slowly added to hydroboration The mol ratio of sodium solution, sodium borohydride and α-ketoglutaric acid is 1:8, with mass fraction be 0.2% hydrochloric acid solution adjust pH to 6 ~ 7,24 h are reacted, mixed solution after must reacting, mixed solution after reaction are poured into mass fraction in 95% ethanol, to stir 1h, making α-ketoglutaric acid modification of chitosan is separated out completely, filters, takes precipitation, and precipitation is washed into 3 ~ 4 with absolute ethyl alcohol, absolute ether successively It is secondary, it is placed in apparatus,Soxhlet's and uses ethanol continuous extraction, dries, obtain α-ketoglutaric acid modification of chitosan powder；
（4）By microbial flocculant crude extract and α-ketoglutaric acid modification of chitosan powder in mass ratio 5:3 is well mixed, quiet Put, pulverized and sieved with the sieve of 200 mesh, collect sieving particle, obtain compound adsorbing metal ions flocculant.
Example 1
Flocculant extracts raw material：The sludge in river is taken, is placed at 4 DEG C and preserves.
No.1 bacterium solid culture medium：According to the mass fraction, 3 parts of beef extract, 6 parts of peptone, 15 parts of agar, NaCl 2 are taken Part, 1000 parts, pH7,121 DEG C sterilizing 20min of water.
No. two mould solid mediums：According to the mass fraction, 20 parts of sucrose, NaNO are taken31 part, K2HPO40.5 part, MgSO40.2 part, FeSO40.01 part, 13 parts of agar, 1000 parts, pH7,115 DEG C sterilizing 20min of water.
No. three actinomyces solid mediums：According to the mass fraction, 18 parts of soluble starch, KNO are taken30.6 part, K2HPO40.3 Part, MgSO4·7H2O0.2 parts, FeSO4·7H2O0.02 parts, NaCl0.4 parts, 15 parts of agar, 1000 parts, pH7,121 DEG C of water go out Bacterium 20min.
No. four fungus solids culture mediums：According to the mass fraction, 180 parts of potato, 20 parts of sucrose, 18 parts of agar, water are taken 1000 parts, pH is naturally, 121 DEG C of sterilizing 20min.
One, two, three, No. four corresponding fluid nutrient mediums：Agar is not added with, other compositions are identical, and corresponding liquid can be made Culture medium.
Fermentation culture：According to the mass fraction, 15 parts of glucose, 0.4 part of urea, 0.2 part of yeast extract, K are taken2HPO43 parts, KH2PO41 part, NaCl0.08 parts, MgSO4·7H2O0.1 parts, (NH4)2SO40.2 part, 1000 parts, pH7.2,115 DEG C sterilizings of water 20min。
The screening of bacterium for producing flocculant of microbe：By purification of bacterial bacterial strain, fungal strain, actinomycetes strain, fungal bacterial strain It is seeded to respectively in fermentation culture（One plant of inoculation is into a nutrient solution）, the quality of every kind of bacterial strain and fermented nutritive liquid Than for 1:50,25 DEG C of shaking table cultures 2 days, centrifugation, take supernatant, obtain strain fermentating liquid by kaolin and water in mass ratio 1: 150 are mixed and made into aqueous suspension ofkaolin, regulated value 8, aqueous suspension ofkaolin, the CaCl that mass fraction is 1%2Solution, strain fermentation Liquid in mass ratio 25:1:1 is well mixed, stirs 5min, mixed liquor is poured into colorimetric cylinder, static 20min, taken under supernatant Liquid at 2cm, its absorbance is determined under 550nm wavelength with spectrophotometer, so as to determine the flocculating rate of every kind of strain, it is determined that XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest.
A kind of preparation method of compound adsorbing metal ions flocculant, comprises the following steps：
（1）Take sludge in mass ratio 1:5 add in the beaker equipped with deionized water, are placed on magnetic stirring apparatus and stir 10min, quiet 1h is put, is centrifuged, supernatant is collected, supernatant is taken into equal four parts in mass ratio 2:9 are seeded to the training of No.1 bacterial liquid respectively Support in base, No. two mould fluid nutrient mediums, No. three actinomyces fluid nutrient mediums, No. four fungi liquid culture mediums, 25 DEG C of cultures 3 My god, by same inoculum concentration, same condition of culture repeats enrichment culture 2 times, obtains No.1, No. two, No. three, No. four enrichment trainings Nutrient solution, No.1, No. two, No. three, No. four enrichment culture liquid are diluted to 10 respectively with the physiological saline that mass fraction is 0.9%-6It is dilute Degree of releasing, No.1, No. two, No. three, No. four enrichment culture liquid after must diluting, respectively streak inoculation are trained in corresponding No.1 bacterial solids Support in base, No. two mould solid mediums, No. three actinomyces solid mediums, No. four fungus solids culture mediums, 25 DEG C of cultures 3 My god, the bacterium bacterial strain that is purified, fungal strain, actinomycetes strain, fungal bacterial strain, by bacterial screening, determine XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest, it is the producing strains of microbial flocculant；
（2）XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 are seeded to No.1 bacterial liquid culture medium, No. two mould liquid trainings respectively Support base, No. three actinomyces fluid nutrient mediums, cultivate in No. four fungi liquid culture mediums, 25 ~ 33 DEG C are cultivated 3 ~ 5 days, obtain XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution, by XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution in mass ratio 2:1:2:2 is mixed Close, obtain mixed bacteria liquid, mixed bacteria liquid is seeded in fermentation culture, 25 DEG C ferment 3 days, take fermentate, centrifuge, take supernatant Liquid, in mass ratio 2 in supernatant:3 add acetone, and 4 DEG C of constant temperature preserve 24h, take precipitation, wash precipitation 2 times with ether, vacuum is done It is dry, obtain microbial flocculant crude extract；
（3）By chitosan and distilled water in mass ratio 1:10 mixed dissolutions, obtain chitosan solution, into chitosan solution add α- The mass ratio of ketoglutaric acid, chitosan solution and α-ketoglutaric acid is 1:2, it is stirred continuously, obtains transparent sticky molten at room temperature Liquid, pH to 4 is adjusted with the sodium hydroxide solution that mass fraction is 0.1 mol/L, stirring reaction 4h, is slowly added to sodium borohydride The mol ratio of solution, sodium borohydride and α-ketoglutaric acid is 1:8, pH to 6 is adjusted with the hydrochloric acid solution that mass fraction is 0.2%, instead 24 h are answered, mixed solution after must reacting, mixed solution after reaction are poured into mass fraction in 95% ethanol, to stir 1h, making α -one Glutaric acid modification of chitosan is separated out completely, filters, takes precipitation, and precipitation is washed 3 times with absolute ethyl alcohol, absolute ether successively, put Ethanol continuous extraction is used in apparatus,Soxhlet's, dries, obtains α-ketoglutaric acid modification of chitosan powder；
（4）By microbial flocculant crude extract and α-ketoglutaric acid modification of chitosan powder in mass ratio 5:3 is well mixed, quiet Put, pulverized and sieved with the sieve of 200 mesh, collect sieving particle, obtain compound adsorbing metal ions flocculant.
Example 2
Flocculant extracts raw material：The sludge in river is taken, is placed at 4 DEG C and preserves.
No.1 bacterium solid culture medium：According to the mass fraction, 5 parts of beef extract, 10 parts of peptone, 20 parts of agar, NaCl are taken 3 parts, 1000 parts, pH7.4,121 DEG C sterilizing 20min of water.
No. two mould solid mediums：According to the mass fraction, 25 parts of sucrose, NaNO are taken32 parts, K2HPO40.8 part, MgSO40.3 part, FeSO40.03 part, 15 parts of agar, 1000 parts, pH7.2,115 DEG C sterilizing 20min of water.
No. three actinomyces solid mediums：According to the mass fraction, 20 parts of soluble starch, KNO are taken31 part, K2HPO40.6 Part, MgSO4·7H2O0.5 parts, FeSO4·7H2O0.03 parts, NaCl0.7 parts, 20 parts of agar, 1000 parts, pH7.4,121 DEG C of water Sterilize 20min.
No. four fungus solids culture mediums：According to the mass fraction, 200 parts of potato, 22 parts of sucrose, 20 parts of agar, water are taken 1000 parts, pH is naturally, 121 DEG C of sterilizing 20min.
One, two, three, No. four corresponding fluid nutrient mediums：Agar is not added with, other compositions are identical, and corresponding liquid can be made Culture medium.
Fermentation culture：According to the mass fraction, 18 parts of glucose, 0.6 part of urea, 0.5 part of yeast extract, K are taken2HPO45 parts, KH2PO42 parts, NaCl0.12 parts, MgSO4·7H2O0.3 parts, (NH4)2SO40.4 part, 1000 parts, pH7.6,115 DEG C sterilizings of water 20min。
The screening of bacterium for producing flocculant of microbe：By purification of bacterial bacterial strain, fungal strain, actinomycetes strain, fungal bacterial strain It is seeded to respectively in fermentation culture（One plant of inoculation is into a nutrient solution）, the quality of every kind of bacterial strain and fermented nutritive liquid Than for 1:50,30 DEG C of shaking table cultures 3 days, centrifugation, take supernatant, obtain strain fermentating liquid by kaolin and water in mass ratio 1: 150 are mixed and made into aqueous suspension ofkaolin, regulated value 9, aqueous suspension ofkaolin, the CaCl that mass fraction is 1%2Solution, strain fermentation Liquid in mass ratio 25:1:1 is well mixed, stirs 5min, mixed liquor is poured into colorimetric cylinder, static 20min, taken under supernatant Liquid at 2cm, its absorbance is determined under 550nm wavelength with spectrophotometer, so as to determine the flocculating rate of every kind of strain, it is determined that XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest.
A kind of preparation method of compound adsorbing metal ions flocculant, comprises the following steps：
（1）Take sludge in mass ratio 1:5 add in the beaker equipped with deionized water, are placed on magnetic stirring apparatus and stir 10min, quiet 2h is put, is centrifuged, supernatant is collected, supernatant is taken into equal four parts in mass ratio 2:9 are seeded to the training of No.1 bacterial liquid respectively Support in base, No. two mould fluid nutrient mediums, No. three actinomyces fluid nutrient mediums, No. four fungi liquid culture mediums, 33 DEG C of cultures 5 My god, by same inoculum concentration, same condition of culture repeats enrichment culture 2 times, obtains No.1, No. two, No. three, No. four enrichment trainings Nutrient solution, No.1, No. two, No. three, No. four enrichment culture liquid are diluted to 10 respectively with the physiological saline that mass fraction is 0.9%-6It is dilute Degree of releasing, No.1, No. two, No. three, No. four enrichment culture liquid after must diluting, respectively streak inoculation are trained in corresponding No.1 bacterial solids Support in base, No. two mould solid mediums, No. three actinomyces solid mediums, No. four fungus solids culture mediums, 33 DEG C of cultures 5 My god, the bacterium bacterial strain that is purified, fungal strain, actinomycetes strain, fungal bacterial strain, by bacterial screening, determine XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest, it is the producing strains of microbial flocculant；
（2）XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 are seeded to No.1 bacterial liquid culture medium, No. two mould liquid trainings respectively Support base, No. three actinomyces fluid nutrient mediums, cultivate in No. four fungi liquid culture mediums, 25 ~ 33 DEG C are cultivated 3 ~ 5 days, obtain XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution, by XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution in mass ratio 2:1:2:2 is mixed Close, obtain mixed bacteria liquid, mixed bacteria liquid is seeded in fermentation culture, 30 DEG C ferment 5 days, take fermentate, centrifuge, take supernatant Liquid, in mass ratio 2 in supernatant:3 add acetone, and 4 DEG C of constant temperature preserve 24h, take precipitation, wash precipitation 3 times with ether, vacuum is done It is dry, obtain microbial flocculant crude extract；
（3）By chitosan and distilled water in mass ratio 1:10 mixed dissolutions, obtain chitosan solution, into chitosan solution add α- The mass ratio of ketoglutaric acid, chitosan solution and α-ketoglutaric acid is 1:2, it is stirred continuously, obtains transparent sticky molten at room temperature Liquid, pH to 5 is adjusted with the sodium hydroxide solution that mass fraction is 0.1 mol/L, stirring reaction 4h, is slowly added to sodium borohydride The mol ratio of solution, sodium borohydride and α-ketoglutaric acid is 1:8, pH to 7 is adjusted with the hydrochloric acid solution that mass fraction is 0.2%, instead 24 h are answered, mixed solution after must reacting, mixed solution after reaction are poured into mass fraction in 95% ethanol, to stir 1h, making α -one Glutaric acid modification of chitosan is separated out completely, filters, takes precipitation, and precipitation is washed 4 times with absolute ethyl alcohol, absolute ether successively, put Ethanol continuous extraction is used in apparatus,Soxhlet's, dries, obtains α-ketoglutaric acid modification of chitosan powder；
（4）By microbial flocculant crude extract and α-ketoglutaric acid modification of chitosan powder in mass ratio 5:3 is well mixed, quiet Put, pulverized and sieved with the sieve of 200 mesh, collect sieving particle, obtain compound adsorbing metal ions flocculant.
Example 3
Flocculant extracts raw material：The sludge in river is taken, is placed at 4 DEG C and preserves.
No.1 bacterium solid culture medium：According to the mass fraction, 4 parts of beef extract, 8 parts of peptone, 17.5 parts of agar, NaCl are taken 2.5 parts, 1000 parts, pH7.2,121 DEG C sterilizing 20min of water.
No. two mould solid mediums：According to the mass fraction, 22.5 parts of sucrose, NaNO are taken31.5 parts, K2HPO40.65 part, MgSO40.25 part, FeSO40.02 part, 14 parts of agar, 1000 parts, pH7,115 DEG C sterilizing 20min of water.
No. three actinomyces solid mediums：According to the mass fraction, 19 parts of soluble starch, KNO are taken30.8 part, K2HPO40.45 part, MgSO4·7H2O0.35 parts, FeSO4·7H2O0.025 parts, NaCl0.55 parts, 17.5 parts of agar, water 1000 Part, pH7.2,121 DEG C of sterilizing 20min.
No. four fungus solids culture mediums：According to the mass fraction, 190 parts of potato, 21 parts of sucrose, 19 parts of agar, water are taken 1000 parts, pH is naturally, 121 DEG C of sterilizing 20min.
One, two, three, No. four corresponding fluid nutrient mediums：Agar is not added with, other compositions are identical, and corresponding liquid can be made Culture medium.
Fermentation culture：According to the mass fraction, 16.5 parts of glucose, 0.5 part of urea, 0.35 part of yeast extract, K are taken2HPO44 Part, KH2PO41.5 parts, NaCl0.1 parts, MgSO4·7H2O0.2 parts, (NH4)2SO40.3 part, 1000 parts, pH7.4,115 DEG C of water goes out Bacterium 20min.
The screening of bacterium for producing flocculant of microbe：By purification of bacterial bacterial strain, fungal strain, actinomycetes strain, fungal bacterial strain It is seeded to respectively in fermentation culture（One plant of inoculation is into a nutrient solution）, the quality of every kind of bacterial strain and fermented nutritive liquid Than for 1:50,27 DEG C of shaking table cultures 2.5 days, centrifugation, take supernatant, obtain strain fermentating liquid by kaolin with water in mass ratio 1:150 are mixed and made into aqueous suspension ofkaolin, regulated value 8.5, aqueous suspension ofkaolin, the CaCl that mass fraction is 1%2Solution, strain Zymotic fluid in mass ratio 25:1:1 is well mixed, stirs 5min, mixed liquor is poured into colorimetric cylinder, static 20min, takes supernatant Liquid at lower 2cm, its absorbance is determined under 550nm wavelength with spectrophotometer, so as to determine the flocculating rate of every kind of strain, really Determine XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest.
A kind of preparation method of compound adsorbing metal ions flocculant, comprises the following steps：
（1）Take sludge in mass ratio 1:5 add in the beaker equipped with deionized water, are placed on magnetic stirring apparatus and stir 10min, quiet 1.5h is put, is centrifuged, supernatant is collected, supernatant is taken into equal four parts in mass ratio 2:9 are seeded to No.1 bacterial liquid respectively In culture medium, No. two mould fluid nutrient mediums, No. three actinomyces fluid nutrient mediums, No. four fungi liquid culture mediums, 29 DEG C of cultures 4 My god, by same inoculum concentration, same condition of culture repeats enrichment culture 2 times, obtains No.1, No. two, No. three, No. four enrichment trainings Nutrient solution, No.1, No. two, No. three, No. four enrichment culture liquid are diluted to 10 respectively with the physiological saline that mass fraction is 0.9%-6It is dilute Degree of releasing, No.1, No. two, No. three, No. four enrichment culture liquid after must diluting, respectively streak inoculation are trained in corresponding No.1 bacterial solids Support in base, No. two mould solid mediums, No. three actinomyces solid mediums, No. four fungus solids culture mediums, 29 DEG C of cultures 4 My god, the bacterium bacterial strain that is purified, fungal strain, actinomycetes strain, fungal bacterial strain, by bacterial screening, determine XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 flocculation activity highest, it is the producing strains of microbial flocculant；
（2）XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 are seeded to No.1 bacterial liquid culture medium, No. two mould liquid trainings respectively Support base, No. three actinomyces fluid nutrient mediums, cultivate in No. four fungi liquid culture mediums, 25 ~ 33 DEG C are cultivated 3 ~ 5 days, obtain XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution, by XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution in mass ratio 2:1:2:2 is mixed Close, obtain mixed bacteria liquid, mixed bacteria liquid is seeded in fermentation culture, 29 DEG C ferment 4 days, take fermentate, centrifuge, take supernatant Liquid, in mass ratio 2 in supernatant:3 add acetone, and 4 DEG C of constant temperature preserve 24h, take precipitation, wash precipitation 3 times with ether, vacuum is done It is dry, obtain microbial flocculant crude extract；
（3）By chitosan and distilled water in mass ratio 1:10 mixed dissolutions, obtain chitosan solution, into chitosan solution add α- The mass ratio of ketoglutaric acid, chitosan solution and α-ketoglutaric acid is 1:2, it is stirred continuously, obtains transparent sticky molten at room temperature Liquid, pH to 4.5 is adjusted with the sodium hydroxide solution that mass fraction is 0.1 mol/L, stirring reaction 4h, is slowly added to hydroboration The mol ratio of sodium solution, sodium borohydride and α-ketoglutaric acid is 1:8, adjust pH extremely with the hydrochloric acid solution that mass fraction is 0.2% 6.5,24 h are reacted, mixed solution after must reacting, mixed solution after reaction are poured into mass fraction in 95% ethanol, to stir 1h, α-ketoglutaric acid modification of chitosan is separated out completely, filter, take precipitation, precipitation is washed 3 with absolute ethyl alcohol, absolute ether successively It is secondary, it is placed in apparatus,Soxhlet's and uses ethanol continuous extraction, dries, obtain α-ketoglutaric acid modification of chitosan powder；
（4）By microbial flocculant crude extract and α-ketoglutaric acid modification of chitosan powder in mass ratio 5:3 is well mixed, quiet Put, pulverized and sieved with the sieve of 200 mesh, collect sieving particle, obtain compound adsorbing metal ions flocculant.
Reference examples：The flocculant of company of Weifang City production.
The flocculant of flocculant obtained by examples detailed above and comparative example is detected, specific detection is as follows：
The 1wt% kaolin aqueous solution is prepared, with the suspension for these flocculants that judge, 100ml suspensions is poured into and carried The volume filled in altogether is in 200ml measuring cylinder.The flocculant and sold flocculant on the market of the present invention is instilled using dropper respectively Identical amount is instilled as sample, after instillation, with plug measuring cylinder, reversion ten times of turning upside down.Then measuring cylinder is made Return to static condition, under ultraviolet specrophotometer, measure supernatant at the wavelength 550nm under absorbance, calculate wadding The flocculating rate and number of viable of solidifying agent.The flocculant prepared is mixed with the solution containing cadmium or copper ion prepared, it is molten containing cadmium Cd in liquid2+Concentration is 100mg/L, Cu in copper-containing solution2+Concentration is 100mg/L, mistake after flocculant addition 10g/L, 2h Filter, with distilled water flushing, test Cd in the aqueous solution2+、Cu2+Concentration.Test result is shown in Table 1.
Flocculating rate %=（Compare supernatant absorption photometric value-sample supernatant absorption photometric value）/ sample supernatant absorption photometric value ×100%
Summary, Flocculating Effect of Flocculant of the invention is good, and absorption heavy metal ability is stronger, is worthy to be popularized.

Claims (6)

1. a kind of compound adsorbing metal ions flocculant preparation method, it is characterised in that the preparation method comprises the following steps：
（1）Take the sludge in river to add in deionized water, be placed on magnetic stirring apparatus and stir, stand, centrifugation, collect supernatant Liquid, take quality equal supernatant four parts are seeded to No.1 bacterial liquid culture medium, No. two mould fluid nutrient mediums, three respectively Cultivated in number actinomyces fluid nutrient medium, No. four fungi liquid culture mediums, by same inoculum concentration, same condition of culture, weight Multiple enrichment culture 2 times, obtains No.1, No. two, No. three, No. four enrichment culture liquid, with the physiological saline that mass fraction is 0.9% by one Number, No. two, No. three, No. four enrichment culture liquid be diluted to 10 respectively-6Dilution factor, No.1, No. two, No. three, No. four richnesses after must diluting Collect nutrient solution, then four kinds of enrichment culture liquid are distinguished into streak inoculation and consolidated in corresponding No.1 bacterium solid culture medium, No. two moulds Cultivated in body culture medium, No. three actinomyces solid mediums, No. four fungus solids culture mediums, the bacterial strain that is purified, mould Bacterial strain, actinomycetes strain, fungal bacterial strain, by bacterial screening, determine XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 as microorganism The producing strains of flocculant；
（2）XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 are seeded to No.1 bacterial liquid culture medium, No. two mould liquid trainings respectively Support base, No. three actinomyces fluid nutrient mediums, culture cultivated in No. four fungi liquid culture mediums, obtain XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solutions, XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution are mixed, mixed bacteria liquid is obtained, mixed bacteria liquid is seeded to Being fermented in fermentation culture, take fermentate, centrifuged, take supernatant, acetone is added in supernatant, constant temperature preserves 24h, takes precipitation, Precipitation is washed with ether 2 ~ 3 times, vacuum drying, obtains microbial flocculant crude extract；
（3）By chitosan and distilled water mixed dissolution, chitosan solution is obtained, α-ketoglutaric acid, room are added into chitosan solution It is stirred continuously under temperature, obtains transparent viscosity solution, the sodium hydroxide solution for being 0.1mol/L with mass fraction adjusts pH to 4 ~ 5, Stirring reaction, sodium borohydride solution is added, pH to 6 ~ 7, reaction, after obtaining reaction are adjusted with the hydrochloric acid solution that mass fraction is 0.2% Mixed solution, mixed solution after reaction is poured into mass fraction to be stirred in 95% ethanol, is separated out, is filtered, take precipitation, will precipitate Washed 3 ~ 4 times with absolute ethyl alcohol, absolute ether successively, be placed in apparatus,Soxhlet's and use ethanol continuous extraction, dried, obtain α -one penta Modified Chitosan powder；
（4）Microbial flocculant crude extract is well mixed with α-ketoglutaric acid modification of chitosan powder, stood, is crushed, mistake Sieve, sieving particle is collected, obtains compound adsorbing metal ions flocculant.
2. the preparation method of compound adsorbing metal ions flocculant according to claim 1, it is characterised in that the step Suddenly（1）Middle sludge and the mass ratio of deionized water are 1:5, four portions of supernatants of equal mass are trained with corresponding No.1 bacterial liquid It is 2 to support base, No. two mould fluid nutrient mediums, No. three actinomyces fluid nutrient mediums, the mass ratio of No. four fungi liquid culture mediums:9.
3. the preparation method of compound adsorbing metal ions flocculant according to claim 1, it is characterised in that the step Suddenly（1）The formula of middle No.1 bacterium solid culture medium is according to the mass fraction, to take 3 ~ 5 parts of beef extract, 6 ~ 10 parts of peptone, agar 15 ~ 20 parts, 2 ~ 3 parts of NaCl, 1000 parts, pH7.2 ± 0.2,121 DEG C sterilizing 20min of water；No. two mould solid mediums are matched somebody with somebody Side is according to the mass fraction, to take 20 ~ 25 parts of sucrose, NaNO31 ~ 2 part, K2HPO40.5 ~ 0.8 part, MgSO40.2 ~ 0.3 part, FeSO40.01 ~ 0.03 part, 13 ~ 15 parts of agar, 1000 parts, pH7.0 ± 0.2,115 DEG C sterilizing 20min of water；No. three actinomyces are consolidated The formula of body culture medium is according to the mass fraction, to take 18 ~ 20 parts of soluble starch, KNO30.6 ~ 1 part, K2HPO40.3 ~ 0.6 part, MgSO4·7H2O0.2 ~ 0.5 part, FeSO4·7H2O0.02 ~ 0.03 part, NaCl0.4 ~ 0.7 part, 15 ~ 20 parts of agar, water 1000 Part, pH7.2 ± 0.2,121 DEG C of sterilizing 20min；The formula of No. four fungus solids culture mediums is according to the mass fraction, to take potato 180 ~ 200 parts, 20 ~ 22 parts of sucrose, 18 ~ 20 parts of agar, 1000 parts of water, pH is naturally, 121 DEG C of sterilizing 20min.
4. the preparation method of compound adsorbing metal ions flocculant according to claim 1, it is characterised in that the step Suddenly（2）The formula of middle fermentation culture is according to the mass fraction, to take 15 ~ 18 parts of glucose, 0.4 ~ 0.6 part of urea, yeast extract 0.2 ~ 0.5 part, K2HPO43 ~ 5 parts, KH2PO41 ~ 2 part, NaCl0.08 ~ 0.12 part, MgSO4·7H2O0.1 ~ 0.3 part, (NH4)2SO40.2 ~ 0.4 part, 1000 parts, pH7.4 ± 0.2,115 DEG C sterilizing 20min of water；One, two, three, the formula of No. four corresponding fluid nutrient mediums To be not added with agar, other compositions are identical, and corresponding fluid nutrient medium can be made.
5. the preparation method of compound adsorbing metal ions flocculant according to claim 1, it is characterised in that the step Suddenly（2）Middle XJ3-14, MJ1-5, FXJ6-9, ZJ2-8 nutrient solution in mass ratio 2:1:2:The quality of 2 mixing, supernatant and acetone Than for 2:3.
6. the preparation method of compound adsorbing metal ions flocculant according to claim 1, it is characterised in that the step Suddenly（3）The mass ratio of middle chitosan and distilled water is 1:10, the mass ratio of chitosan solution and α-ketoglutaric acid is 1:2, boron hydrogen The mol ratio for changing sodium and α-ketoglutaric acid is 1:8.
The preparation method of compound adsorbing metal ions flocculant according to claim 1, it is characterised in that the step （4）The mass ratio of middle microbial flocculant crude extract and α-ketoglutaric acid modification of chitosan powder is 5:3.
CN201710977173.2A 2017-10-19 2017-10-19 A kind of preparation method of compound adsorbing metal ions flocculant Pending CN107686172A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108793426A (en) * 2018-06-19 2018-11-13 深圳文科园林股份有限公司 A kind of microbial flocculant compound preparation method and its application in water treatment field
CN109292934A (en) * 2018-11-20 2019-02-01 好太阳（沈阳）环境治理有限公司 Environment-friendly novel flocculating purifying agent
CN117550723A (en) * 2023-11-07 2024-02-13 石家庄华滋生物工程有限公司 Microorganism-loaded sewage treatment agent and preparation method thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108793426A (en) * 2018-06-19 2018-11-13 深圳文科园林股份有限公司 A kind of microbial flocculant compound preparation method and its application in water treatment field
CN109292934A (en) * 2018-11-20 2019-02-01 好太阳（沈阳）环境治理有限公司 Environment-friendly novel flocculating purifying agent
CN117550723A (en) * 2023-11-07 2024-02-13 石家庄华滋生物工程有限公司 Microorganism-loaded sewage treatment agent and preparation method thereof

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