CN108410769A - Fresh water complex microorganism bottom changes the preparation method of agent - Google Patents
Fresh water complex microorganism bottom changes the preparation method of agent Download PDFInfo
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Abstract
The invention discloses the preparation methods that fresh water complex microorganism bottom changes agent, including：Prepare fermentation medium, fermentation, absorption, crosslinking, wheatfeed, maize cob meal, starch wastewater and fermentation synergist are mixed into fermentation medium first, rhodococcus erythropolis, S. cervisiae and the mixed bacteria liquid of bacillus subtilis are accessed and carries out fermented and cultured and purifies to obtain microbial flocculant, then sesame stalk activated carbon, enzyme solution, microbial flocculant and microorganism formulation are adsorbed, then changes agent with cross-linking agents up to fresh water complex microorganism bottom.It has the beneficial effect that：Preparation method is simple for this, the yield of polysaccharide and yield are higher in preparation process, the mechanical strength and high stability of final product, mass-transfer performance is preferable with balling property, fresh water complex microorganism bottom changes agent and is improved to aquaculture wastewater by flocculation, enzyme degradation, the potent effect of three kinds of microbial degradation, its Biodegradable is comprehensive, safe, nontoxic and non-secondary pollution.
Description
Technical field
The present invention relates to cultivation fresh water bottoms to change agent field, changes the preparation side of agent more particularly, to fresh water complex microorganism bottom
Method.
Technical background
Pond is both the environment of cultured fishes and the growth of other aquatiles and the Treatment stations of its secretion, excreta
Institute, after largely being fed during breeding production, the excrement of residual bait and aquatile, corpse, dead algae constantly increase
Add and cannot exclude outside pond, be deposited on bottom of pond, one layer of black mud is formed in pond bottom.Organic matter in mud is in anoxic
Under the conditions of fermentation decompose generate largely be unfavorable for fish and aquatile growth substance, such as ammonia, hydrogen sulfide, methane, hydrogen, have
Machine acid, mercaptan, rudimentary amine etc., these substances not only directly endanger cultured fishes and aquatile, but also can make entire pond
The water quality deterioration of environment, pH value reduce, and to influence the metabolism and growth and development of breed variety, feed coefficient are caused to increase
Greatly, aquaculture cost increases, or even breed variety is caused to be poisoned to death, and heavy losses are caused to the economic benefit of culture-cycle.
Invention content
The purpose of the present invention is to provide the preparation method that a kind of fresh water complex microorganism bottom changes agent, this preparation method is simple
Easy, the yield of polysaccharide and yield are higher in preparation process, and final product bottom changes the mechanical strength and high stability of agent, mass transfer performances
Can be preferable with balling property, fresh water complex microorganism bottom changes agent and passes through flocculation, enzyme degradation, the potent effect pair of three kinds of microbial degradation
Aquaculture wastewater is improved, and Biodegradable is comprehensive, safe, nontoxic and non-secondary pollution.
Used microorganism and its purchase enterprise are in the method for the present invention：Raw biochemical reagents are ground in rhodococcus erythropolis, Shanghai
Co., Ltd；S. cervisiae, Yantai Man Sen commerce and trade Co., Ltd；Bacillus subtilis, saccharomycete, the Cangzhou City macro animal of benefit protect
Jian Pin Co., Ltds；Chlorella, the Xi'an bio tech ltd Tian Guangyuan；Actinomyces, Yangzhou Haicheng biotechnology are limited
Company；Bafillus natto, the Guangzhou bio tech ltd Lv Hui.
The present invention is directed to the problem of being mentioned in background technology, and the technical solution taken is：Fresh water complex microorganism bottom changes agent
Preparation method, including：Fermentation medium, fermentation, absorption, crosslinking are prepared, following steps are specifically included：
Prepare fermentation medium：It is auxiliary material by major ingredient, 30~50 parts of maize cob meals of 200~220 parts of wheatfeeds, separately adds 1.5
~2.5 parts of honey, 3~4 portions of sucrose, addition starch wastewater to dry matter content are 45~55%, and 1.2~1.5 parts of hairs are added after sterilizing
Ferment synergist is up to fermentation medium；Fermentation medium is prepared using starch wastewater and wheatfeed, maize cob meal, is not only realized
Waste utilization protects environment, and also achieves the increment of material, and higher moisture is conducive to nutriment and is in culture medium
Institute of Micro-biology absorbs, and can promote microorganism growth, the progress of enzymatic reaction and distributing for fermentation heat production, to improve fermentation
Efficiency and metabolin yield；
Fermentation：Seed mixture liquid is inoculated with by the 10.8~12.2% of fermentation medium dry weight in fermentation medium, stirring is equal
It is even, it is placed in 35~37 DEG C, ferment in the constant incubator under the conditions of 150~160r/min 60~72h；Take zymotic fluid in 6000~
8000r/min, 10~30min is centrifuged under the conditions of 2~4 DEG C, collect supernatant and the acetone of 3~5 times of volumes is sufficiently mixed, 24~
Centrifugation purification is carried out using the same terms after 36h, the sediment obtained after centrifugation is dried in vacuo, obtains microbial flocculant；
By enzyme effect caused by microorganism in wheat bran substrate, contribute to the release of bran polysaccharide, makes more bran polysaccharides no longer
Fettered by various firm chemical bonds, make its direct separate out, without excessively high temperature damage chemical bond so that its
It is detached from；When sewage disposal, under the conditions of existing for flocculation aid calcium chloride, calcium ion is by Coulomb attraction by negatively charged colloid
Particle furthers, and forms calcium ion-colloidal solid conjugate therewith, and then reduces the thickness of the diffusion electric double layer of colloidal solid
Degree, furthered the distance between colloidal solid, a kind of bridging agent of microbial flocculant picture, mainly by its functional group and calcium from
Two or more calcium ions-colloidal solid conjugate is adsorbed onto on flocculant macromolecular chain by the ionic bond between son,
So as to complete the flocculation of colloidal solid, calcium ion-colloidal solid conjugate is adsorbed onto on a microbial flocculant strand
While, it can be adsorbed onto on other flocculating agent molecule chains, and then generate a kind of floccule body of tridimensional network, floccule body exists
Quickly net is caught and volume sweeps colloidal solid during rapid sedimentation, is finally reached ideal flocculating effect；
Absorption：By sesame stalk activated carbon, enzyme solution, microbial flocculant and microorganism formulation according to mass ratio 100:300~
500:15~50:1.5~1.8 uniformly mix and are adsorbed, and adsorption temp is 22~24 DEG C, and 3~5h of thermostatic absorption obtains compound
Enzyme solution；Sesame stalk activated carbon is porous structure, and mass-transfer performance is stronger, with enzyme solution, microbial flocculant, microorganism formulation energy
The contact for enough generating larger area can greatly improve appendicular activity and degradation efficiency；
Crosslinking：It is 1 by mass ratio:3~5:1~2:5~8:12~20 polyethylene glycol, sodium alginate, calcium chloride, deionized water
Mixed solvent is mixed to obtain with complex enzyme liquid, the wherein molecular weight of polyethylene glycol is a concentration of the 0.85 of 500~1200, sodium alginate
A concentration of 0.2~0.4mol/L of~1.05mol/L, calcium chloride；Then according to weight ratio 1:1.8~2.2 are added dropwise to solvent
In crosslinking agent, controlled at 0~1 DEG C, 5~30min of stirring crosslinking in ice-water bath, then move to drying at room temperature 45~
I.e. obtain immobilized microorganism bottom changes agent microballoon to 60min, and obtained immobilized microorganism bottom is changed agent microballoon to be rushed with physiological saline
It washes 3~5 times, as fresh water complex microorganism bottom changes agent；The mechanical strength that complex microorganism bottom changes agent is higher, resistance to physical impact,
Mass-transfer performance is higher with balling property, at the same complex microorganism bottom change agent stability it is very high, be not easily broken decomposition, Ke Yixun
Speed is expeditiously flocculated, wastewater pollutants of degrading, and nontoxic secondary residual, cost is relatively low, is suitble to large-scale production and application, is one
The safe and efficient fresh water bottom of kind changes agent.
Preferably, it is 1 that seed mixture liquid, which is weight ratio,:1~3:5~10 rhodococcus erythropolis, S. cervisiae with it is withered
The mixed bacteria liquid of careless bacillus；It selects mixed fungus fermentation that can play the synergistic effect between a variety of strains, single bacterium hair can be reduced
The disadvantage of ferment, there may be synbiosis, metabolite, and complementary effect occurs, intermediate product can be overcome excessive between a variety of strains
To how glycogenetic unfavorable factor, therefore be conducive to the fermentation of bran polysaccharide.
Preferably, fermentation synergist includes the substance of following content：55~80% urea, 2~10% α~methyl α-naphthyl acetate
Sodium, 12~18% calcium oxide, 2.5~5.5% magnesium sulfate, 1.2~1.5% dimethyl phosphate；Ammoniation can occur for urea,
Wheat bran, the maize cob meal porosity after ammoniated treatment increase, and increase with the contact area of enzyme, are conducive to the generation of polysaccharide；Hair
There is the α-naphthaleneacidsodium Yu dimethyl phosphate of special proportioning synergistic effect, the synergistic effect can promote withered grass in ferment synergist
Bacillus enhances the secretion to acetyl coenzyme A, and then promotes its condensation reaction with oxaloacetic acid, accelerates the conjunction of citric acid
At can increase substantially the speed of bacillus subtilis tricarboxylic acid cycle, accelerate the growth and breeding and hair of bacillus subtilis
Ferment is metabolized so that more enzymes are acted on fermentation substrate, and then improve the yield of polysaccharide.
Preferably, the preparation process of enzyme solution is：It chooses decontamination scene and filters out the chlorella with higher decontamination performance
It is cultivated, by 7~15d is cultivated and the bead algae solution in exponential phase filters, and filter membrane selects 0.35~0.45 μ
M removes the filter membrane containing chlorella, and 3~4 times of PBS buffer solution is added in liquid nitrogen frozen later, and the pH of PBS buffer solution is 7.4
~7.5, a concentration of 0.4~0.5mol/L；Be ground at a temperature of 1~2 DEG C, lapping liquid pour into centrifuge tube and 1~2 DEG C,
15~30min is centrifuged under conditions of 10000~12000r/min, is taken out supernatant up to enzyme solution, is put in 1~2 DEG C of preservation；From except
For extraction enzyme solution as immobilised enzymes, specific aim is stronger, while can be according to heterogeneity in the chlorella that dirty scene filters out
Sewage selects different algae solutions, and chlorella producing enzyme is efficient, while institute's producing enzyme has higher degradability to pollutant, under low temperature
Extraction enzyme solution will not impact enzyme, have higher practicability.
Preferably, it is 1 that crosslinking agent, which is mass ratio,:0.02~0.04:0.2~0.4:8~15 methyl orthosilicate, 3,5-
Difluoro-benzoic acid, three oxygen methyl-monosilane of methyl and methanol, mix to obtain crosslinking agent in ice-water bath；Two of 3,5- difluoro-benzoic acids
Fluorine atom has larger elecrtonegativity, and the base of a fruit that can strengthen sugar chain closes, and enhances bridging property, it is micro- that raising immobilized microorganism bottom changes agent
The mechanical strength and stability of ball enhance impact resistance, extend the service life that bottom changes agent microballoon, reduce cost, resistance to physics punching
Hit, improve bottom and change the mass-transfer performance and balling property of agent microballoon, at the same immobilized microorganism bottom change agent microballoon stability it is very high,
It is not easily broken decomposition, can rapidly, expeditiously degrade flocculation, degradation of contaminant, and nontoxic secondary residual, cost is relatively low, is suitble to rule
Modelling produces and application, is that a kind of safe and efficient fresh water bottom changes agent.
Preferably, it is 1 that microorganism formulation, which is weight ratio,:0.5~0.8:0.3~0.5 actinomyces, saccharomycete, natto
The mixed microorganism preparation of bacillus；Bottom changes agent and the flocculation of the pollutants such as protein, starch, fat in polluted-water is reunited
After be settled down to the bottom, during sedimentation with after the completion of sedimentation, immobilized microorganism bottom change enzyme preparation in agent microballoon with it is micro-
Actinomyces, saccharomycete, bafillus natto in biological agent can degrade to organic pollution, be dropped by flocculation, enzyme
Solution, the potent effect of three kinds of microbial degradation improve aquaculture wastewater, and Biodegradable is comprehensive, safe, nontoxic nothing
Evil and non-secondary pollution.
Compared with the prior art, the advantages of the present invention are as follows：
1）The α-naphthaleneacidsodium Yu dimethyl phosphate of special proportioning have synergistic effect, the synergistic effect can be in fermentation synergist
Promote bacillus subtilis to enhance the secretion to acetyl coenzyme A, and then it is accelerated to be condensed synthesizing citric acid with oxaloacetic acid, substantially
Degree improves the speed of bacillus subtilis tricarboxylic acid cycle, accelerates the growth and breeding and fermentating metabolism of bacillus subtilis so that
More enzymes are acted on fermentation substrate, and then improve the yield of polysaccharide；
2）Two fluorine atoms of 3, the 5- difluoro-benzoic acids in crosslinking agent have larger elecrtonegativity, can strengthen the base of a fruit of sugar chain
It closes, enhances bridging property, improve mechanical strength and stability that immobilized microorganism bottom changes agent microballoon, enhance impact resistance, extend
Bottom changes the service life of agent microballoon, reduces cost, and resistance to physical impact improves mass-transfer performance and balling property that bottom changes agent microballoon；
3）Fresh water complex microorganism bottom, which changes after the pollutant flocculation in polluted-water is reunited in agent, is settled down to the bottom, in the mistake of sedimentation
After the completion of Cheng Zhongyu sedimentations, bottom changes the microorganism formulation in enzyme preparation and microorganism formulation in agent microballoon can be to organic contamination
Object is degraded, and is improved to aquaculture wastewater by flocculation, enzyme degradation, the potent effect of three kinds of microbial degradation, biology point
Comprehensive, safe, the nontoxic and non-secondary pollution of solution property.
Specific implementation mode
The present invention program is described further below by embodiment：
Embodiment 1：
Fresh water complex microorganism bottom changes the preparation method of agent, includes the following steps：
1）It is auxiliary material by major ingredient, 30 parts of maize cob meals of 200 parts of wheatfeeds, separately adds 1.5 parts of honey, 3 portions of sucrose, adds starch
Waste water to dry matter content is 45%, and 1.2 parts of fermentation synergists are added after sterilizing up to fermentation medium；Under fermentation synergist includes
The substance of row content：75% urea, 5% α-naphthaleneacidsodium, 14% calcium oxide, 4.8% magnesium sulfate, 1.2% phosphoric acid diformazan
Ester；2）Seed mixture liquid is inoculated with by the 10.8% of fermentation medium dry weight in fermentation medium, seed mixture liquid is weight ratio
It is 1:1:5 rhodococcus erythropolis, S. cervisiae and the mixed bacteria liquid of bacillus subtilis, stirs evenly, be placed in 35 DEG C,
Ferment 60h in constant incubator under the conditions of 150r/min；It takes zymotic fluid to centrifuge 10min under the conditions of 6000r/min, 2 DEG C, receives
Collection supernatant and the acetone of 3 times of volumes are sufficiently mixed, and centrifugation purification are carried out using the same terms afterwards for 24 hours, by what is obtained after centrifugation
Sediment is dried in vacuo, and obtains microbial flocculant；3）By sesame stalk activated carbon, enzyme solution, microbial flocculant and microorganism
Preparation is according to mass ratio 100:300:15:1.5 uniformly mix and are adsorbed, and microorganism formulation is that weight ratio is 1:0.5:0.3
Actinomyces, saccharomycete, bafillus natto mixed microorganism preparation, adsorption temp be 22 DEG C, thermostatic absorption 3h obtains compound
Enzyme solution；4）It is 1 by mass ratio:3:1:5:12 polyethylene glycol, sodium alginate, calcium chloride, deionized water are mixed with complex enzyme liquid
Mixed solvent, the wherein molecular weight of polyethylene glycol is 500~1200, a concentration of 0.85mol/L of sodium alginate, calcium chloride
A concentration of 0.2mol/L；Then according to weight ratio 1:1.8 are added dropwise to solvent in crosslinking agent, and crosslinking agent is that mass ratio is 1:
0.02:0.2:8 methyl orthosilicate, 3,5- difluoro-benzoic acids, three oxygen methyl-monosilane of methyl and methanol, controlled at 0 DEG C,
Stirring crosslinking 5min in ice-water bath, then moves to drying at room temperature 45min and obtains immobilized microorganism bottom and change agent microballoon, will obtain
Immobilized microorganism bottom change agent microballoon normal saline flushing 3 times, as fresh water complex microorganism bottom changes agent.
Embodiment 2：
Fresh water complex microorganism bottom changes the preparation method of agent, includes the following steps：
1）Prepare fermentation medium：It is auxiliary material by major ingredient, 50 parts of maize cob meals of 220 parts of wheatfeeds, it is another to add 2.5 parts of honey, 4
Part sucrose, addition starch wastewater to dry matter content are 55%, and 1.5 parts of fermentation synergists are added after sterilizing up to fermentation medium；Profit
Fermentation medium is prepared with starch wastewater and wheatfeed, maize cob meal, waste utilization is not only realized, protects environment, and
The increment of material is also achieved, higher moisture is conducive to nutriment as Institute of Micro-biology's intake in culture medium, can promote micro-
Biological growth, the progress of enzymatic reaction and distributing for fermentation heat production, to improve the efficiency of fermentation and the yield of metabolin；
2）Fermentation：Seed mixture liquid is inoculated with by the 12.2% of fermentation medium dry weight in fermentation medium, stirs evenly, sets
Ferment 72h in constant incubator under the conditions of 37 DEG C, 160r/min；Zymotic fluid is taken to be centrifuged under the conditions of 8000r/min, 4 DEG C
30min, collects supernatant and the acetone of 5 times of volumes is sufficiently mixed, and carries out centrifugation purification using the same terms after 36h, will centrifuge
The sediment vacuum drying obtained afterwards, obtains microbial flocculant；By enzyme effect caused by microorganism in wheat bran substrate, have
The release for helping bran polysaccharide makes more bran polysaccharides no longer be fettered by various firm chemical bonds, keeps it directly free
Out, without excessively high temperature damage chemical bond to disengage it from；
3）Absorption：By sesame stalk activated carbon, enzyme solution, microbial flocculant and microorganism formulation according to mass ratio 100:500:
50:1.8 uniformly mix and are adsorbed, and adsorption temp is 24 DEG C, and thermostatic absorption 5h obtains complex enzyme liquid；Sesame stalk activated carbon is
Porous structure, mass-transfer performance is stronger, and the contact of larger area can be generated with enzyme solution, microbial flocculant, microorganism formulation,
Appendicular activity and degradation efficiency can greatly be improved；
4）Crosslinking：It is 1 by mass ratio:5:2:8:20 polyethylene glycol, sodium alginate, calcium chloride, deionized water and complex enzyme liquid
Mixed solvent is mixed to obtain, the wherein molecular weight of polyethylene glycol is 500~1200, a concentration of 1.05mol/L of sodium alginate, chlorination
A concentration of 0.4mol/L of calcium；Then according to weight ratio 1:2.2 are added dropwise to solvent in crosslinking agent, controlled at 1 DEG C, in ice
Stirred in water bath is crosslinked 30min, then moves to drying at room temperature 60min and obtains immobilized microorganism bottom and changes agent microballoon, will obtain
Immobilized microorganism bottom change agent microballoon normal saline flushing 5 times, as fresh water complex microorganism bottom changes agent；Complex microorganism
The mechanical strength that bottom changes agent is higher, and resistance to physical impact, mass-transfer performance is higher with balling property, while complex microorganism bottom changes agent
Stability is very high, is not easily broken decomposition, can rapidly, expeditiously flocculate, wastewater pollutants of degrading, nontoxic secondary residual, cost
It is relatively low, it is suitble to large-scale production and application, is that a kind of safe and efficient fresh water bottom changes agent.
Seed mixture liquid is that weight ratio is 1:3:10 rhodococcus erythropolis, S. cervisiae and the mixing of bacillus subtilis
Bacterium solution.
Fermentation synergist includes the substance of following content：78% urea, 3% α-naphthaleneacidsodium, 15% calcium oxide, 2.5%
Magnesium sulfate, 1.5% dimethyl phosphate.
The preparation process of enzyme solution is：Choosing decontamination scene and filter out, there is the chlorella of higher decontamination performance to be cultivated,
By 15d is cultivated and the bead algae solution in exponential phase filters, and filter membrane selects 0.45 μm, removes containing chlorella
Filter membrane, liquid nitrogen frozen are added 4 times of PBS buffer solution, are ground at a temperature of 2 DEG C later, lapping liquid pour into centrifuge tube and
2 DEG C, 30min is centrifuged under conditions of 12000r/min, take out supernatant up to enzyme solution, be put in 2 DEG C of preservations.
Crosslinking agent is that mass ratio is 1:0.04:0.4:15 methyl orthosilicate, 3,5- difluoro-benzoic acids, three oxygen methyl of methyl
Silane and methanol mix to obtain crosslinking agent in ice-water bath.
Microorganism formulation is that weight ratio is 1:0.8:0.5 actinomyces, the mixed microorganism of saccharomycete, bafillus natto
Preparation.
Embodiment 3：
Fresh water complex microorganism bottom changes the preparation method of agent, including：Fermentation medium, fermentation, absorption, crosslinking are prepared, it is specific to wrap
Include following steps：
Prepare fermentation medium：It is auxiliary material by major ingredient, 40 parts of maize cob meals of 200 parts of wheatfeeds, separately adds 2 parts of honey, 3 parts of sugarcanes
Sugar, addition starch wastewater to dry matter content are 50%, and 1.2 parts of fermentation synergists are added after sterilizing up to fermentation medium；Utilize shallow lake
Powder waste water prepares fermentation medium with wheatfeed, maize cob meal, not only realizes waste utilization, protects environment, but also real
The increment of material is showed, higher moisture is conducive to nutriment as Institute of Micro-biology's intake in culture medium, can promote microorganism
Growth, the progress of enzymatic reaction and distributing for fermentation heat production, to improve the efficiency of fermentation and the yield of metabolin；
Fermentation：Seed mixture liquid is inoculated with by the 12% of fermentation medium dry weight in fermentation medium, stirs evenly, is placed in 37
DEG C, ferment in the constant incubator under the conditions of 150r/min 60h；Zymotic fluid is taken to be centrifuged under the conditions of 6500r/min, 2 DEG C
25min, collects supernatant and the acetone of 4 times of volumes is sufficiently mixed, and carries out centrifugation purification using the same terms after 36h, will centrifuge
The sediment vacuum drying obtained afterwards, obtains microbial flocculant；By enzyme effect caused by microorganism in wheat bran substrate, have
The release for helping bran polysaccharide makes more bran polysaccharides no longer be fettered by various firm chemical bonds, keeps it directly free
Out, without excessively high temperature damage chemical bond to disengage it from；When sewage disposal, the item existing for flocculation aid calcium chloride
Under part, calcium ion is furthered negatively charged colloidal solid by Coulomb attraction, and is formed calcium ion-colloidal solid therewith and combined
Object, and then the thickness of the diffusion electric double layer of colloidal solid is reduced, furthered the distance between colloidal solid, microbial flocculant
As a kind of bridging agent, mainly by the ionic bond between its functional group and calcium ion by two or more calcium ions-
Colloidal solid conjugate is adsorbed onto on flocculant macromolecular chain, so as to complete the flocculation of colloidal solid, calcium ion-colloidal solid
While conjugate is adsorbed onto on a microbial flocculant strand, it can be adsorbed onto on other flocculating agent molecule chains, in turn
A kind of floccule body of tridimensional network is generated, floccule body during rapid sedimentation quickly catch and volume sweeps colloidal solid by net,
It is finally reached ideal flocculating effect；
Absorption：By sesame stalk activated carbon, enzyme solution, microbial flocculant and microorganism formulation according to mass ratio 100:400:25:
1.6 uniformly mix and are adsorbed, and adsorption temp is 24 DEG C, and thermostatic absorption 4h obtains complex enzyme liquid；Sesame stalk activated carbon is more
Pore structure, mass-transfer performance is stronger, and the contact of larger area can be generated with enzyme solution, microbial flocculant, microorganism formulation, can
Greatly to improve appendicular activity and degradation efficiency；
Crosslinking：It is 1 by mass ratio:4:1:7:13 polyethylene glycol, sodium alginate, calcium chloride, deionized water and complex enzyme liquid is mixed
Close to obtain mixed solvent, wherein the molecular weight of polyethylene glycol be 500~1200, a concentration of 1mol/L of sodium alginate, calcium chloride
A concentration of 0.3mol/L；Then according to weight ratio 1:2 are added dropwise to solvent in crosslinking agent, controlled at 1 DEG C, in ice-water bath
Stirring crosslinking 25min, then moves to drying at room temperature 45min and obtains immobilized microorganism bottom and change agent microballoon, the fixation that will be obtained
Change microorganism bottom and change agent microballoon normal saline flushing 4 times, as fresh water complex microorganism bottom changes agent；Complex microorganism bottom changes agent
Mechanical strength it is higher, resistance to physical impact, mass-transfer performance is higher with balling property, while complex microorganism bottom changes the stability of agent
It is very high, it is not easily broken decomposition, can rapidly, expeditiously be flocculated, wastewater pollutants of degrading, nontoxic secondary residual, cost is relatively low, fits
Large-scale production and application are closed, is that a kind of safe and efficient fresh water bottom changes agent.
Seed mixture liquid is that weight ratio is 1:2:7 rhodococcus erythropolis, S. cervisiae and the mixing of bacillus subtilis
Bacterium solution；It selects mixed fungus fermentation that can play the synergistic effect between a variety of strains, the disadvantage of single bacterium fermentation, a variety of strains can be reduced
Between there may be synbiosis, complementary effect occurs for metabolite, the excessively multipair polysaccharide of intermediate product can be overcome to generate unfavorable
Factor, therefore be conducive to the fermentation of bran polysaccharide.
Fermentation synergist includes the substance of following content：70% urea, 10% α-naphthaleneacidsodium, 15% calcium oxide, 3.8%
Magnesium sulfate, 1.2% dimethyl phosphate；Ammoniation, wheat bran, the maize cob meal porosity after ammoniated treatment can occur for urea
Increase, increases with the contact area of enzyme, be conducive to the generation of polysaccharide；In fermentation synergist the α-naphthaleneacidsodium of special proportioning with
There is dimethyl phosphate synergistic effect, the synergistic effect bacillus subtilis can be promoted to enhance the secretion to acetyl coenzyme A, into
And promote its condensation reaction with oxaloacetic acid, accelerate the synthesis of citric acid, bacillus subtilis tricarboxylic can be increased substantially
The speed of acid cycle, accelerates the growth and breeding and fermentating metabolism of bacillus subtilis so that and more enzymes are acted on fermentation substrate,
And then improve the yield of polysaccharide.
The preparation process of enzyme solution is：Choosing decontamination scene and filter out, there is the chlorella of higher decontamination performance to be cultivated,
By 10d is cultivated and the bead algae solution in exponential phase filters, and filter membrane selects 0.4 μm, removes containing chlorella
4 times of PBS buffer solution is added in filter membrane, liquid nitrogen frozen later, and the pH of PBS buffer solution is 7.4, a concentration of 0.4mol/L；At 2 DEG C
At a temperature of be ground, lapping liquid pours into centrifuge tube and centrifuges 20min under conditions of 2 DEG C, 10000r/min, take out supernatant
Up to enzyme solution, it is put in 2 DEG C of preservations；Extraction enzyme solution is as immobilised enzymes, specific aim out of decontamination scene filters out chlorella
It is relatively strong, while different algae solutions can be selected according to heterogeneity sewage, chlorella producing enzyme is efficient, while institute's producing enzyme is to pollution
Object has higher degradability, and enzyme will not be impacted by extracting enzyme solution under low temperature, have higher practicability.
Crosslinking agent is that mass ratio is 1:0.02:0.2:10 methyl orthosilicate, 3,5- difluoro-benzoic acids, three oxygen methyl of methyl
Silane and methanol mix to obtain crosslinking agent in ice-water bath；Two fluorine atoms of 3,5- difluoro-benzoic acids have larger elecrtonegativity,
The base of a fruit that sugar chain can be strengthened closes, and enhances bridging property, improves mechanical strength and stability that immobilized microorganism bottom changes agent microballoon, increases
Strong impact resistance extends the service life that bottom changes agent microballoon, reduces cost, and resistance to physical impact improves the mass transfer that bottom changes agent microballoon
Performance and balling property, at the same immobilized microorganism bottom change agent microballoon stability it is very high, be not easily broken decomposition, can it is rapid,
Expeditiously degradation flocculation, degradation of contaminant, nontoxic secondary residual, cost is relatively low, is suitble to large-scale production and application, is a kind of peace
Entirely, efficient fresh water bottom changes agent.
Microorganism formulation is that weight ratio is 1:0.6:0.4 actinomyces, the mixed microorganism of saccharomycete, bafillus natto
Preparation；Bottom, which changes after the flocculation of the pollutants such as protein, starch, fat in polluted-water is reunited in agent, is settled down to the bottom, is settling
During with after the completion of sedimentation, immobilized microorganism bottom change actinomyces in enzyme preparation and microorganism formulation in agent microballoon,
Saccharomycete, bafillus natto can degrade to organic pollution, three kinds strong by flocculation, enzyme degradation, microbial degradation
Effect effect improves aquaculture wastewater, and Biodegradable is comprehensive, safe, nontoxic and non-secondary pollution.
The physicochemical property that fresh water complex microorganism bottom in Examples 1 to 3 is changed to agent compares, result such as 1 institute of table
Show, as shown in Table 1, fresh water complex microorganism bottom change agent mechanical strength and immobilization degree it is higher, the mass-transfer performance of microballoon with
Balling-up is preferable, has higher degradation rate to the organic pollution in freshwater aquiculture water.
Fresh water complex microorganism bottom in 1. Examples 1 to 3 of table changes the physicochemical property of agent
| Project | Mechanical strength/mN | Mass-transfer performance/min | Balling-up | Immobilization efficiency/% | Degradation rate/% |
| Embodiment 1 | 81.5 | 26 | Very well | 91.3 | 85.6 |
| Embodiment 2 | 84.8 | 22 | Very well | 94.8 | 89.2 |
| Embodiment 3 | 80.2 | 27 | Preferably | 92.6 | 84.7 |
Routine operation in operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (9)
1. fresh water complex microorganism bottom changes the preparation method of agent, including：Fermentation medium, fermentation, absorption, crosslinking are prepared, it is special
Sign is：It is added in the preparation fermentation medium step and contains the following fermentation synergist containing quantity of material：55~80% urine
Element, 2~10% α-naphthaleneacidsodium, 12~18% calcium oxide, 2.5~5.5% magnesium sulfate, 1.2~1.5% dimethyl phosphate,
Total content is 100%.
2. fresh water complex microorganism according to claim 1 bottom changes the preparation method of agent, it is characterised in that：The preparation hair
Ferment culture medium step is：It is auxiliary material by major ingredient, 30~50 parts of maize cob meals of 200~220 parts of wheatfeeds, separately adds 1.5~2.5
Part honey, 3~4 portions of sucrose, addition starch wastewater to dry matter content are 45~55%, and 1.2~1.5 parts of fermentation increasings are added after sterilizing
Agent is imitated up to fermentation medium.
3. fresh water complex microorganism according to claim 1 bottom changes the preparation method of agent, it is characterised in that：The fermentation step
Suddenly it is：Seed mixture liquid is inoculated with ferment at constant temperature in fermentation medium by the 10.8~12.2% of fermentation medium dry weight, is fermented
It takes that supernatant is collected by centrifugation in zymotic fluid and the acetone of 3~5 times of volumes is sufficiently mixed after the completion, stands after 24~36h using identical
Condition carries out centrifugation purification, and the sediment obtained after centrifugation is dried in vacuo up to microbial flocculant.
4. fresh water complex microorganism according to claim 3 bottom changes the preparation method of agent, it is characterised in that：The fermentation step
In rapid, seed mixture liquid is that weight ratio is 1:1~3:5~10 rhodococcus erythropolis, S. cervisiae and bacillus subtilis
Mixed bacteria liquid.
5. fresh water complex microorganism according to claim 1 bottom changes the preparation method of agent, it is characterised in that：The absorption step
Suddenly it is：By sesame stalk activated carbon, enzyme solution, microbial flocculant and microorganism formulation according to mass ratio 100:300~500:15
~50:1.5~1.8 uniformly mix and are adsorbed, and adsorption temp is 22~24 DEG C, and 3~5h of thermostatic absorption obtains complex enzyme liquid.
6. fresh water complex microorganism according to claim 5 bottom changes the preparation method of agent, it is characterised in that：The absorption step
In rapid, the preparation method of enzyme solution is：Choosing decontamination scene and filter out, there is the chlorella of higher decontamination performance to be cultivated, and will train
It supports 7~15d and the bead algae solution in exponential phase is filtered, remove the filter membrane containing chlorella and liquid nitrogen frozen,
3~4 times of PBS buffer solution is added later, is centrifuged after grinding, takes out supernatant up to enzyme solution.
7. fresh water complex microorganism according to claim 5 bottom changes the preparation method of agent, it is characterised in that：The absorption step
It is 1 that microorganism formulation in rapid, which is weight ratio,:0.5~0.8:0.3~0.5 actinomyces, saccharomycete, bafillus natto it is mixed
Close microorganism formulation.
8. fresh water complex microorganism according to claim 1 bottom changes the preparation method of agent, it is characterised in that：The crosslinking step
Suddenly it is：It is 1 by mass ratio:3~5:1~2:5~8:12~20 polyethylene glycol, sodium alginate, calcium chloride, deionized water with it is multiple
Synthase liquid mixes to obtain mixed solvent, then according to weight ratio 1:1.8~2.2 are added dropwise to solvent in crosslinking agent, controlled at 0
It~1 DEG C, 5~30min of stirring crosslinking, then moves to drying at room temperature and obtains immobilized microorganism bottom and change agent microballoon, physiological saline
It is that fresh water complex microorganism bottom changes agent after rinsing well.
9. fresh water complex microorganism according to claim 8 bottom changes the preparation method of agent, it is characterised in that：The crosslinking agent
It is 1 for mass ratio:0.02~0.04:0.2~0.4:8~15 methyl orthosilicate, 3,5- difluoro-benzoic acids, three oxygen methyl of methyl
The mixture of silane and methanol mixes to obtain crosslinking agent in ice-water bath.
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| CN109534513A (en) * | 2018-12-28 | 2019-03-29 | 厦门理工学院 | The bacterium algae immobilization preparation purified in situ aquiculture waste water method that charcoal is strengthened |
| CN109804962A (en) * | 2019-01-26 | 2019-05-28 | 江西新农匠生物技术有限公司 | A kind of aquatic products healthy ecology cultivation water adjusting method |
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| CN109534513A (en) * | 2018-12-28 | 2019-03-29 | 厦门理工学院 | The bacterium algae immobilization preparation purified in situ aquiculture waste water method that charcoal is strengthened |
| CN109804962A (en) * | 2019-01-26 | 2019-05-28 | 江西新农匠生物技术有限公司 | A kind of aquatic products healthy ecology cultivation water adjusting method |
| CN109804962B (en) * | 2019-01-26 | 2021-08-24 | 北京新农匠生态科技有限公司 | Water transfer method for healthy ecological aquaculture |
| CN111470720A (en) * | 2020-04-16 | 2020-07-31 | 南昌航空大学 | Method for denitrifying rare earth ore wastewater by using supported microalgae |
| CN112320969A (en) * | 2020-10-19 | 2021-02-05 | 辽宁科技学院 | Preparation method of traditional Chinese medicine wastewater treatment agent based on microorganisms |
| CN113072156A (en) * | 2021-04-12 | 2021-07-06 | 济宁鑫惠生水产养殖专业合作社 | Aquaculture tail water treatment agent, preparation process and preparation device |
| CN113830979A (en) * | 2021-11-25 | 2021-12-24 | 深圳市华研环保科技有限公司 | Microbial treatment method for aquaculture sewage |
| CN115716671A (en) * | 2022-12-08 | 2023-02-28 | 山东珺宜环保科技有限公司 | Composite microbial flocculant and preparation method and application thereof |
| CN115716671B (en) * | 2022-12-08 | 2023-09-26 | 山东珺宜环保科技有限公司 | Composite microbial flocculant and preparation method and application thereof |
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