CN108034689A - A kind of microbial flocculant and its application in the rich phosphorus sewage of processing - Google Patents
A kind of microbial flocculant and its application in the rich phosphorus sewage of processing Download PDFInfo
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- CN108034689A CN108034689A CN201711066945.3A CN201711066945A CN108034689A CN 108034689 A CN108034689 A CN 108034689A CN 201711066945 A CN201711066945 A CN 201711066945A CN 108034689 A CN108034689 A CN 108034689A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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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
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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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
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/347—Use of yeasts or fungi
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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
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- 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
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
Abstract
A kind of application the invention discloses microbial flocculant and its in the rich phosphorus sewage of processing.The microbial flocculant is prepared by following methods：(1) flcos producing bacteria is inoculated with crude fibre raw material, one grade fermemtation matrix is made, carries out aerobic fermentation, obtains one grade fermemtation thing；(2) the inoculation solution fiber clostridium ATCC35319 in one grade fermemtation thing, carries out anaerobic fermentation, obtains second order fermentation thing；(3) after the water content of second order fermentation thing is down to 20~30%, slime bacteria is inoculated with, three grade fermemtation matrix is made；(4) aerobic fermentation is carried out, obtains three grade fermemtation thing；(5) three grade fermemtation thing is extracted using ethyl acetate, obtains the microbial flocculant；The flcos producing bacteria includes aspergillus niger CGMCC3.3289 and streptomyces parvus CGMCC4.5927.The present invention ferments crude fibre raw material using flcos producing bacteria Clostridium baratii slime bacteria successively, and the flocculant of acquisition has excellent phosphor-removing effect.
Description
Technical field
The present invention relates to rich phosphorus technical field of sewage, and in particular to a kind of microbial flocculant and its in the rich phosphorus of processing
Application in sewage.
Background technology
COD and SS are not only sought out during municipal sewage treatment at present, and dephosphorization and denitrogenation are proposed
Higher requirement.And in rural area, because environmental consciousness is thin, because seek it is cheap and use rich phosphorus washing powder and there are plenty of such people, this
The phosphorus pollution condition for resulting in rural area river water also allows of no optimist.
Sewage dephosphorization technology can be divided into two class of biological phosphate-eliminating and chemical dephosphorization.Wherein, the essence of biological phosphate-eliminating is to pass through
Phosphorus and accumulation of the polyP bacteria under aerobic condition in excess ingestion waste water in the cell, are then discharged as excess sludge.Usually
Think that Biological Phosphorus Removal System operating cost is low, management is convenient, but treatment effect is unstable, and polyP bacteria has fixed growth in itself
In the cycle, be difficult to realize long-time stable qualified discharge, and when phosphate concn of especially intaking is higher, such case is particularly evident.
And chemical dephosphorization is then phosphate is separated in the form of chemical precipitation from sewage by adding chemical agent.
The characteristics of chemical phosphorus removal system is maximum is can to control phosphor-removing effect by controlling added amount of chemical, but chemical agent is usual takes
With higher, its application in Practical Project is limited.
The content of the invention
This application provides a kind of microbial flocculant, which has excellent phosphor-removing effect.
A kind of microbial flocculant, is prepared by following methods：
(1) flcos producing bacteria is inoculated with crude fibre raw material, it is the one grade fermemtation base that 50~70%, pH is 3~6 that water content, which is made,
Matter, carries out aerobic fermentation, obtains one grade fermemtation thing；
(2) inoculation solution fiber clostridium (Clostridium cellulolyticum) ATCC35319 in one grade fermemtation thing,
Anaerobic fermentation is carried out, obtains second order fermentation thing；
(3) after the water content of the second order fermentation thing is down to 20~30%, slime bacteria is inoculated with second order fermentation thing, is made
Into water content below 40%, the three grade fermemtation matrix that pH is 6~8；
(4) aerobic fermentation is carried out, obtains three grade fermemtation thing；
(5) three grade fermemtation thing is extracted using ethyl acetate, takes leaching liquor, filter to take clear liquid, it is dry, described in acquisition
Microbial flocculant；
The flcos producing bacteria includes：
①：Geneva Mucor (Mucor genevensis) CGMCC 3.4901, trichoderma reesei (Trichoderma
Reesei) CGMCC 3.3711, annulus aspergillus (Aspergillus zonatus) CGMCC3.4469, aspergillus niger
(Aspergillus niger) CGMCC3.3289, beamforming mould (Penicillium isariiforme) CGMCC3.4025 and
At least one of Rhizopus oryzae (Rhizopus oryzae) CGMCC 3.5818；
And 2.：Light ash streptomycete (Streptomyces spororaveus) CGMCC4.5873 of spore, streptomyces parvus
(Streptomyces parvus) CGMCC4.5927 and Streptomyces Luteogriseus (Streptomyces luteogriseus)
At least one of CGMCC4.2001.
The present invention carries out biology turn using 2. actinomyces coordinated that the fungi 1. organized and the are organized to crude fibre raw material
Change, metabolin of the generation with flocculation phosphor-removing effect, antagonism is not present between two groups of microorganisms listed by the present invention, each
Synergy is there is likely to be between caused metabolin, the microbial flocculant of the present invention is imitated with more preferable dephosphorization
Fruit.
The present invention carries out crude fibre raw material first with above-mentioned flcos producing bacteria the bioconversion of the first round, but aerobic by a wheel
After fermentation, flcos producing bacteria has all come into production cycle end (decaying the phase) substantially；Therefore the present invention is in one grade fermemtation thing
Continue inoculation solution fiber clostridium, solution fiber clostridium can also generate the metabolism with flocculation phosphor-removing effect in anaerobic fermentation process
Thing, and between the metabolin that produces of flcos producing bacteria, between the metabolin that solution fiber clostridium produces, metabolin that flcos producing bacteria produces with
It also occur that complicated chemical reaction between the metabolin that solution fiber clostridium produces so that in the microbial flocculant finally obtained
Contain a variety of functional groups.
Similarly, it is also limited to the bioconversion of crude fibre raw material to solve fiber clostridium.Therefore the present invention further exists
Be inoculated with slime bacteria in second order fermentation thing, slime bacteria can using in second order fermentation thing remaining crude fibre raw material as source of nutrition,
Or can dead using in second order fermentation thing or active relatively low flcos producing bacteria conciliate fiber clostridium thalline as source of nutrition, into one
Step generation can make the metabolin of the calcium phosphate precipitation in sewage, substantially increase raw material availability and capacity efficiency.
Preferably, the flcos producing bacteria includes：Aspergillus niger (Aspergillus niger) CGMCC3.3289 and small strepto-
Bacterium (Streptomyces parvus) CGMCC4.5927.
In step (1), in terms of dry weight, the inoculum concentration of the flcos producing bacteria is 104~106A spore/gram crude fibre raw material.Connect
After kind, the water content and pH that adjust one grade fermemtation matrix can provide more suitable propagation environment for flcos producing bacteria.
Before flcos producing bacteria is inoculated with, first crude fibre raw material can be crushed, degree of grinding is more thin better.So can not only
Crude fibre utilization rate is enough improved, and also allows for flcos producing bacteria, solution fiber clostridium and slime bacteria growth, during subsequent extracted flocculant,
Also higher extraction efficiency can be obtained.
Crude fibre raw material can be crops, such as stalk, sweet potato dregs, bagasse etc..
Preferably, in step (1), the temperature of the aerobic fermentation is 28~35 DEG C, and the aerobic fermentation time is 25~30
My god.
Preferably, in terms of dry weight, described solution fiber clostridium (Clostridium cellulolyticum) ATCC35319
Inoculum concentration be 108~1010CFU/ grams of crude fibre raw material.
In step (2), the time of the anaerobic fermentation is 50~70 days.The longer anaerobic fermentation time can be metabolin
Between occur chemical reaction provide time enough.
In the present invention, the slime bacteria includes：Orange-yellow myxococcus (Myxococcus xanthus) CGMCC1.3865 and
Sorangium cellulosum (Soranguim cellulosum) ATCC25532.Wherein, orange-yellow myxococcus can be with one grade fermemtation thing
The residuum of flcos producing bacteria as nutriment, and sorangium cellulosum then can using in one grade fermemtation thing remaining crude fibre raw material as
Nutriment, both, which share out the work and help one another, improves raw material availability, additionally it is possible to produces a large amount of mucus, glycoprotein, viscous more is contained in mucus
The macromoleculars such as sugar, these macromoleculars and the metabolin and reactant in second order fermentation thing, or mixing, or react to each other, collaboration increases
Further improve to effect the flocculation ability and dephosphorization efficiency of flocculant.
Preferably, in terms of dry weight, orange-yellow myxococcus (Myxococcus xanthus) CGMCC1.3865 and fibre
The inoculum concentration for tieing up heap capsule bacterium (Soranguim cellulosum) ATCC25532 is 109~1011CFU/ grams of crude fibre raw material.
Preferably, in step (3), the temperature of the aerobic fermentation is 30~35 DEG C, and fermentation time is 45~55 days.
Present invention also offers application of the microbial flocculant in the rich phosphorus sewage of processing.
Preferably, the dosage of the microbial flocculant is 0~0.5g/L richness phosphorus sewage, i.e. 0 ＜ dosages≤
0.5g/L richness phosphorus sewage.
When carrying out rich phosphorus sewage disposal, the dosage of microbial flocculant can according to the concentration of phosphate from sewage and
Make corresponding adjust.For the microbial flocculant of the present invention, when the concentration of phosphate from sewage is in 5mg/L or so, dosage
In 0.1g/L, phosphatic concentration is smaller at this time, and the dosage of microbial flocculant of the invention can also be reduced accordingly.
When the concentration of phosphate from sewage is in 200mg/L or so, dosage is in 0.04~0.05g/L.The microorganism of the present invention
Flocculant has excellent phosphor-removing effect, using less dosage can efficient process richness phosphorus sewage, make in rich phosphorus sewage
Phosphorus content is reduced to normal level.
Compared with prior art, beneficial effects of the present invention are embodied in：
(1) present invention carries out crude fibre raw material using 2. actinomyces coordinated that the fungi 1. organized and the are organized biological
Conversion, generation have the metabolin of flocculation phosphor-removing effect, antagonism are not present between two groups of microorganisms listed by the present invention, respectively
Synergy is there is likely to be between caused metabolin, the microbial flocculant of the present invention is imitated with more preferable dephosphorization
Fruit；
(2) present invention relays continued access kind solution fiber clostridium in one grade fermemtation thing, solves fiber clostridium in anaerobic fermentation process,
Also can generate the metabolin with flocculation phosphor-removing effect, and between the metabolin that produces of flcos producing bacteria, solution fiber clostridium produces
Also occur that complicated chemistry is anti-between metabolin, between the metabolin that flcos producing bacteria produces and the metabolin that solution fiber clostridium produces
Should so that contain a variety of functional groups in the microbial flocculant finally obtained.
(3) present invention is further inoculated with slime bacteria in second order fermentation thing, and slime bacteria can be to remain in second order fermentation thing
Crude fibre raw material as source of nutrition, or fiber can be conciliate with dead in second order fermentation thing or active relatively low flcos producing bacteria
Clostridium thalline can make the metabolin of the calcium phosphate precipitation in sewage, substantially increase original as source of nutrition, further generation
Expect utilization rate and capacity efficiency.
Brief description of the drawings
Fig. 1 is treatment effect of the microbial flocculant to the phosphorous simulated wastewater of various concentrations of each embodiment preparation；
Fig. 2 be embodiment 1 prepare microbial flocculant under different dosages to the treatment effect of phosphorous simulated wastewater；
Middle flocculant in microbial flocculant and embodiment 1 that Fig. 3 is prepared for comparative example 1~4 is to phosphorous simulation
The treatment effect of waste water.
Embodiment
Technical scheme is described in further detail with reference to the accompanying drawings and detailed description.
Solution fiber clostridium (Clostridium cellulolyticum) ATCC35319 and fiber stack used in the present invention
Capsule bacterium (Soranguim cellulosum) ATCC25532 is from American Type Culture collection warehousing (American type
Culture collection) purchase acquisition, remaining bacterial strain is from China General Microbiological culture presevation administrative center
What (China General Microbiological Culture Collection Center, CGMCC) purchase obtained.
Embodiment 1
A kind of microbial flocculant of the present embodiment, is prepared by following preparation method：
1st, activated spawn
(1) aspergillus niger (Aspergillus niger) CGMCC3.3289 is inoculated in sterilized bran mass,
Cultivated 5 days at 28 DEG C, treat the spore quantity of aspergillus niger in culture more than 107During a/gram culture, black-koji mould is obtained
Agent, it is stand-by.
(2) that streptomyces parvus (Streptomyces parvus) CGMCC4.5927 is inoculated in sterilized Gause I is oblique
In the culture medium of face, after being cultivated 7 days at 30 DEG C, the sterile water with bead is injected into test tube slant, fully after vibration, filtering
Into monospore suspension, spore count is detected using photoelectric turbidimetry, and spore quantity is made as 109The spore suspension of a/mL, is treated
With.
(3) solution fiber clostridium (Clostridium cellulolyticum) ATCC35319 is inoculated in sterilized dress
In the 250mL anaerobism bottles for having 100mL DCB-1 culture mediums (crushing maize stalk containing 5.0g/L), 35 DEG C are cultivated 3 days, are collected
Thalline, is made concentration as 1011The bacteria suspension of CFU/mL, it is stand-by.
(4) orange-yellow myxococcus (Myxococcus xanthus) CGMCC1.3865 is inoculated in sterilized VY/2 to train
Support in base, cultivated 7 days at 35 DEG C, collect gloeospore, concentration is made as 1013The gloeospore suspension of a gloeospore/mL, it is stand-by.
(5) sorangium cellulosum (Soranguim cellulosum) ATCC25532 is inoculated in sterilized CNST solids
(media surface lays the maize straw and bagasse 1 for having crushing in culture medium:1 mixture), cultivate 7 days, receive at 35 DEG C
Collect gloeospore, concentration is made as 1013The gloeospore suspension of a gloeospore/mL, it is stand-by.
2nd, microbial flocculant is prepared
(1) fresh corn stalk and each 10kg of bagasse are taken, is fitted into after pulverizer pulverization process in mixer, side stirring
While add water to crude fibre raw material water content for 55% or so when, nitric acid is added in a manner of spraying, treats the pH of crude fibre raw material extremely
During 3-4, continue plus water is until the water content of crude fibre raw material is 60%；Add above-mentioned aspergillus niger microbial inoculum 40g, above-mentioned streptomyces parvus
Spore suspension 100mL, after stirring evenly, obtains one grade fermemtation matrix；
(2) one grade fermemtation matrix is transferred to four walls to be equipped with the plastic box of air hole, plastic box is placed in 28
Constant temperature aerobic fermentation is carried out in DEG C, after 28 days, obtains one grade fermemtation thing；
(3) the one grade fermemtation thing in case is transferred in a plastic barrel, adds above-mentioned solution fiber clostridium (Clostridium
Cellulolyticum) ATCC35319 bacteria suspensions 200mL；Compacting, ensures to fill one grade fermemtation thing in plastic barrel, bung hole covers tightly
Shut, plastic barrel is placed at 32 DEG C and carries out constant-temperatureanaerobic anaerobic fermentation 60 days, obtains second order fermentation product (water content 25%)；
(4) pH of second order fermentation product is adjusted to after 7~7.5, adds the gloeospore of the above-mentioned orange-yellow myxococcus of 200mL
The gloeospore suspension of suspension and the above-mentioned sorangium cellulosums of 200mL (Soranguim cellulosum) ATCC25532, stirring are equal
It is even, it is transferred to four walls and is equipped with the plastic box of air hole, plastic box is placed in 32 DEG C and carries out constant temperature aerobic fermentation, 50
Three grade fermemtation thing is obtained after it；
(5) three grade fermemtation thing is taken out, 50L ethyl acetate is added into three grade fermemtation thing, is extracted at room temperature, every time
Extract 24 it is small when, extract 6~7 times repeatedly, collect leaching liquor, filter to take clear liquid, rotate to dry, obtain the microorganism of the present embodiment
Flocculant.
In fermentation process, part primary fermentate, second order fermentation thing are taken respectively, adds appropriate ethyl acetate thereto,
Extracted at room temperature, when extraction 24 is small every time, extract 6~7 times repeatedly, collect leaching liquor, filter to take clear liquid, rotated to dry,
Flocculant among level-one, flocculant among two level are obtained respectively, it is spare.
Embodiment 2
A kind of microbial flocculant of the present embodiment, is prepared by following preparation method：
1st, activated spawn
(1) beamforming mould (Penicillium isariiforme) CGMCC 3.4025 is inoculated in sterilized wheat bran
In culture medium, cultivated 5 days at 28 DEG C, treat the spore quantity of beamforming mould in culture more than 107During a/gram culture, obtain
Beamforming mould microbial inoculum is obtained, it is stand-by.
(2) Streptomyces Luteogriseus (Streptomyces luteogriseus) CGMCC4.2001 is inoculated in sterilized
In Gause I slant medium, after being cultivated 7 days at 30 DEG C, the sterile water with bead is injected into test tube slant, fully
After vibration, monospore suspension is filtered into, spore count is detected using photoelectric turbidimetry, and spore quantity is made as 109The spore of a/mL
Sub- suspension, it is stand-by.
(3) solution fiber clostridium (Clostridium cellulolyticum) ATCC35319 is inoculated in sterilized dress
In the 250mL anaerobism bottles for having 100mL DCB-1 culture mediums (crushing maize stalk containing 5.0g/L), 35 DEG C are cultivated 3 days, are collected
Thalline, is made concentration as 1011The bacteria suspension of CFU/mL, it is stand-by.
(4) orange-yellow myxococcus (Myxococcus xanthus) CGMCC1.3865 is inoculated in sterilized VY/2 to train
Support in base, cultivated 7 days at 35 DEG C, collect gloeospore, concentration is made as 1013The gloeospore suspension of a gloeospore/mL, it is stand-by.
(5) sorangium cellulosum (Soranguim cellulosum) ATCC25532 is inoculated in sterilized CNST solids
(media surface lays the maize straw and bagasse 1 for having crushing in culture medium:1 mixture), cultivate 7 days, receive at 35 DEG C
Collect gloeospore, concentration is made as 1013The gloeospore suspension of a gloeospore/mL, it is stand-by.
2nd, microbial flocculant is prepared
(1) fresh corn stalk and each 10kg of bagasse are taken, is fitted into after pulverizer pulverization process in mixer, side stirring
While add water to crude fibre raw material water content for 45% or so when, nitric acid is added in a manner of spraying, treats the pH of crude fibre raw material extremely
During 3-4, continue plus water is until the water content of crude fibre raw material is 50%；Add above-mentioned beamforming mould microbial inoculum 30g, above-mentioned gamboge ash
Streptomycete spore suspension 80mL, after stirring evenly, obtains one grade fermemtation matrix；
(2) one grade fermemtation matrix is transferred to four walls to be equipped with the plastic box of air hole, plastic box is placed in 28
Constant temperature aerobic fermentation is carried out in DEG C, after 25 days, obtains one grade fermemtation thing；
(3) the one grade fermemtation thing in case is transferred in a plastic barrel, adds above-mentioned solution fiber clostridium (bacteria suspension 160mL；
Compacting, ensures to fill one grade fermemtation thing in plastic barrel, and bung hole, which covers tightly, to be shut, and plastic barrel is placed in progress constant-temperatureanaerobic anaerobic hair at 32 DEG C
Ferment 50 days, obtains second order fermentation product (water content 20%)；
(4) pH of second order fermentation product is adjusted to after 7~7.5, adds the gloeospore of the above-mentioned orange-yellow myxococcus of 180mL
The gloeospore suspension of suspension and the above-mentioned sorangium cellulosums of 180mL, stirs evenly, and is transferred to the modeling that four walls are equipped with air hole
In hopper, plastic box is placed in progress constant temperature aerobic fermentation in 32 DEG C, three grade fermemtation thing is obtained after 45 days；
(5) three grade fermemtation thing is taken out, 50L ethyl acetate is added into three grade fermemtation thing, is extracted at room temperature, every time
Extract 24 it is small when, extract 6~7 times repeatedly, collect leaching liquor, filter to take clear liquid, rotate to dry, obtain the microorganism of the present embodiment
Flocculant.
Embodiment 3
A kind of microbial flocculant of the present embodiment, is prepared by following preparation method：
1st, activated spawn
(1) annulus aspergillus (Aspergillus zonatus) CGMCC3.4469 is inoculated in sterilized bran mass
In, cultivated 5 days at 28 DEG C, treat the spore quantity of annulus aspergillus in culture more than 107During a/gram culture, annulus is obtained
Aspergillus microbial inoculum, it is stand-by.
(2) light ash streptomycete (Streptomyces spororaveus) CGMCC4.5873 of spore is inoculated in sterilized
In Gause I slant medium, after being cultivated 7 days at 30 DEG C, the sterile water with bead is injected into test tube slant, fully
After vibration, monospore suspension is filtered into, spore count is detected using photoelectric turbidimetry, and spore quantity is made as 109The spore of a/mL
Sub- suspension, it is stand-by.
(3) solution fiber clostridium (Clostridium cellulolyticum) ATCC35319 is inoculated in sterilized dress
In the 250mL anaerobism bottles for having 100mL DCB-1 culture mediums (crushing maize stalk containing 5.0g/L), 35 DEG C are cultivated 3 days, are collected
Thalline, is made concentration as 1011The bacteria suspension of CFU/mL, it is stand-by.
(4) orange-yellow myxococcus (Myxococcus xanthus) CGMCC1.3865 is inoculated in sterilized VY/2 to train
Support in base, cultivated 7 days at 35 DEG C, collect gloeospore, concentration is made as 1013The gloeospore suspension of a gloeospore/mL, it is stand-by.
(5) sorangium cellulosum (Soranguim cellulosum) ATCC25532 is inoculated in sterilized CNST solids
(media surface lays the maize straw and bagasse 1 for having crushing in culture medium:1 mixture), cultivate 7 days, receive at 35 DEG C
Collect gloeospore, concentration is made as 1013The gloeospore suspension of a gloeospore/mL, it is stand-by.
2nd, microbial flocculant is prepared
(1) fresh corn stalk and each 10kg of bagasse are taken, is fitted into after pulverizer pulverization process in mixer, side stirring
While add water to crude fibre raw material water content for 65% or so when, nitric acid is added in a manner of spraying, treats the pH of crude fibre raw material extremely
During 3-4, continue plus water is until the water content of crude fibre raw material is 70%；Add above-mentioned annulus aspergillus microbial inoculum 55g, the above-mentioned light ash of spore
Streptomycete spore suspension 150mL, after stirring evenly, obtains one grade fermemtation matrix；
(2) one grade fermemtation matrix is transferred to four walls to be equipped with the plastic box of air hole, plastic box is placed in 28
Constant temperature aerobic fermentation is carried out in DEG C, after 30 days, obtains one grade fermemtation thing；
(3) the one grade fermemtation thing in case is transferred in a plastic barrel, adds above-mentioned solution fiber clostridium bacteria suspension 320mL；
Compacting, ensures to fill one grade fermemtation thing in plastic barrel, and bung hole, which covers tightly, to be shut, and plastic barrel is placed in progress constant-temperatureanaerobic anaerobic hair at 32 DEG C
Ferment 70 days, obtains second order fermentation product (water content 30%)；
(4) pH of second order fermentation product is adjusted to after 7~7.5, adds the gloeospore of the above-mentioned orange-yellow myxococcus of 350mL
The gloeospore suspension of suspension and the above-mentioned sorangium cellulosums of 350mL (Soranguim cellulosum) ATCC25532, stirring are equal
It is even, it is transferred to four walls and is equipped with the plastic box of air hole, plastic box is placed in 32 DEG C and carries out constant temperature aerobic fermentation, 55
Three grade fermemtation thing is obtained after it；
(5) three grade fermemtation thing is taken out, 50L ethyl acetate is added into three grade fermemtation thing, is extracted at room temperature, every time
Extract 24 it is small when, extract 6~7 times repeatedly, collect leaching liquor, filter to take clear liquid, rotate to dry, obtain the microorganism of the present embodiment
Flocculant.
The phosphor-removing effect of 4 microbial flocculant of embodiment
1st, microbial flocculant is to phosphatic removal effect in simulated wastewater
21, beaker is taken, is divided into three groups, pouring into 100mL potassium dihydrogen phosphates respectively in every group of beaker, (concentration is successively
For 5mg/L, 10mg/L, 20mg/L, 40mg/L, 80mg/L, 100mg/L, 200mg/L), thrown respectively into first group of each beaker
Enter the dephosphorization microbial flocculant of the preparation of 0.01g embodiments 1, putting into 0.01g embodiments 2 respectively into second group of each beaker makes
Standby dephosphorization microbial flocculant, the dephosphorization microorganism wadding of the preparation of 0.01g embodiments 3 is put into the 3rd group of each beaker respectively
Solidifying agent, stirs evenly, and in 100r/min, vibrates mixing 40min at room temperature, centrifugation, goes to precipitate, and takes supernatant, detects supernatant
The concentration of middle potassium dihydrogen phosphate, calculates removal efficiency of the dephosphorization microbial flocculant to potassium dihydrogen phosphate of each embodiment, as a result
See Fig. 1.
As seen from Figure 1, the dephosphorization microbial flocculant that prepared by each embodiment is equal to the phosphorous simulated wastewater that concentration is 5mg/L
With more than 98% treatment effeciency, processing of the dephosphorization microbial flocculant that wherein prepared by embodiment 1 to phosphorous simulated wastewater
Effect is best, still has 92.3% dephosphorization efficiency, phosphorous simulated wastewater when phosphorus concentration is 20mg/L in phosphorous simulated wastewater
Still there is 63.7% dephosphorization efficiency when middle phosphorus concentration is 200mg/L.
2nd, influence of the microbial flocculant dosage to phosphatic removal effect in simulated wastewater
21 beakers are taken, are divided into seven groups, the potassium dihydrogen phosphate that 100mL concentration is 200mg/L is poured into every beaker
Solution put into as simulated wastewater, respectively into six groups of beakers each 0.01g, 0.02g of microbial flocculant prepared by embodiment 1,
0.03g, 0.04g, 0.05g, 0.06g, 0.07g, stir evenly, and in 100r/min, vibrate mixing 40min at room temperature, centrifugation, goes
Precipitation, takes supernatant, detects the concentration of potassium dihydrogen phosphate in supernatant, calculates each group dephosphorization microbial flocculant to biphosphate
The removal efficiency of potassium, is averaged, and the result is shown in Fig. 2.
From Figure 2 it can be seen that with the increase of dosage, the treatment effeciency of the microbial flocculant of embodiment 1 to simulated wastewater
Also gradually step up, when dosage is 0.04g, treatment effeciency reaches 87.3%, continues after increasing dosage, treatment effeciency carries
Between lift-off less.
3rd, phosphor-removing effect of the microbial flocculant to sanitary sewage
Dongyang City Wu Ning streets river river water 200mL is taken, detection wherein phosphate concn is 0.6mg/L, by the river water
It is divided into two parts, puts into microbial flocculant prepared by 0.01g, 0.005g embodiment 1 into two parts of river waters respectively, stir evenly,
In 100r/min, mixing 40min is vibrated at room temperature, and centrifugation, goes to precipitate, and takes supernatant, detects potassium dihydrogen phosphate in supernatant
Concentration.
Detect that phosphatic concentration is each about 0.0006mg/L in two parts of river waters, treatment effeciency reaches 99%.
It can be seen from the experimental result of embodiment 4 when phosphorus content is relatively low in simulated wastewater, it can add on a small quantity
The microbial flocculant of the present invention, adds be obtained with good phosphor-removing effect on a small quantity.When the phosphorus content in simulated wastewater
When higher, it can suitably increase the dosage of microbial flocculant.
Comparative example 1
Microbial flocculant is prepared using method same as Example 1, but step " 2, prepare microbial flocculant it
(1) " in, it is added without streptomyces parvus spore suspension.
Comparative example 2
Microbial flocculant is prepared using method same as Example 1, but step " 2, prepare microbial flocculant it
(1) " in, it is added without aspergillus niger microbial inoculum.
Comparative example 3
Microbial flocculant is prepared using method same as Example 1, but step " 2, prepare microbial flocculant it
(4) " in, it is added without the gloeospore suspension of orange-yellow myxococcus.
Comparative example 4
Microbial flocculant is prepared using method same as Example 1, but step " 2, prepare microbial flocculant it
(4) " in, it is added without the gloeospore suspension of sorangium cellulosum.
Flocculant and two level centre flocculant among level-one in Example 1, and micro- life prepared by comparative example 1~4
Thing flocculant；18, beaker is taken again, is divided into six groups, pours into the biphosphate that 100mL concentration is 5mg/L respectively in every group of beaker
Potassium solution, flocculant among 0.01g level-ones is put into first group of each beaker, 0.01g is put into second group of each beaker
Flocculant among two level, 1 microbial flocculant of 0.01g comparative examples is put into the 3rd group of each beaker, to the 4th group of each burning
2 microbial flocculant of 0.01g comparative examples is put into cup, 3 microbial flocculation of 0.01g comparative examples is put into the 5th group of each beaker
Agent, 4 microbial flocculant of 0.01g comparative examples is put into the 6th group of each beaker；Stir evenly, in 100r/min, at room temperature
Vibration mixing 40min, centrifugation, goes to precipitate, and takes supernatant, detects the concentration of potassium dihydrogen phosphate in supernatant, calculates each flocculant
To the removal efficiency of potassium dihydrogen phosphate, the result is shown in Fig. 3.
As seen from Figure 3, the dephosphorization efficiency of flocculant among level-one can be improved by solving the anaerobic fermentation that fiber clostridium carries out, and
The aerobic fermentation that orange-yellow myxococcus (comparative example 4) or sorangium cellulosum (comparative example 3) individually carry out can further improve two
The dephosphorization efficiency of flocculant among level, this is because orange-yellow myxococcus decompose thalline, sorangium cellulosum in second order fermentation thing
Remaining cellulose raw material in second order fermentation thing is decomposed, not only improves raw material availability, but also improve the dephosphorization efficiency of flocculant.
But orange-yellow myxococcus and sorangium cellulosum are carried out at the same time castering action maximum of the fermentation to dephosphorization efficiency, prompt
There may be there may be synergy between the metabolin of collaboration symbiosis, or both between the two for this.
And the dephosphorization efficiency of the flocculant obtained when streptomyces parvus (comparative example 2) or aspergillus niger (comparative example 1) individualism
The dephosphorization efficiency of flocculant obtained in the presence of at the same time without both is high, indicates this between the two there may be collaboration symbiosis,
Or both metabolin between there may be synergy.
Claims (9)
1. a kind of microbial flocculant, it is characterised in that be prepared by following methods：
(1) flcos producing bacteria is inoculated with crude fibre raw material, it is the one grade fermemtation matrix that 50~70%, pH is 3~6 that water content, which is made,
Aerobic fermentation is carried out, obtains one grade fermemtation thing；
(2) inoculation solution fiber clostridium (Clostridium cellulolyticum) ATCC35319 in one grade fermemtation thing, carries out
Anaerobic fermentation, obtains second order fermentation thing；
(3) after the water content of the second order fermentation thing is down to 20~30%, slime bacteria is inoculated with second order fermentation thing, is made and contains
The three grade fermemtation matrix that water is below 40%, pH is 6~8；
(4) aerobic fermentation is carried out, obtains three grade fermemtation thing；
(5) three grade fermemtation thing is extracted using ethyl acetate, takes leaching liquor, filter to take clear liquid, it is dry, obtain micro- life
Thing flocculant；
The flcos producing bacteria includes：
①：Geneva Mucor (Mucor genevensis) CGMCC 3.4901, trichoderma reesei (Trichoderma reesei)
CGMCC 3.3711, annulus aspergillus (Aspergillus zonatus) CGMCC3.4469, aspergillus niger (Aspergillus
Niger) CGMCC3.3289, beamforming mould (Penicillium isariiforme) CGMCC 3.4025 and Rhizopus oryzae
At least one of (Rhizopus oryzae) CGMCC 3.5818；
And 2.：Light ash streptomycete (Streptomyces spororaveus) CGMCC4.5873 of spore, streptomyces parvus
(Streptomyces parvus) CGMCC4.5927 and Streptomyces Luteogriseus (Streptomyces luteogriseus)
At least one of CGMCC4.2001.
2. microbial flocculant as claimed in claim 1, it is characterised in that the flcos producing bacteria includes：Aspergillus niger
(Aspergillus niger) CGMCC3.3289 and streptomyces parvus (Streptomyces parvus) CGMCC4.5927.
3. microbial flocculant as claimed in claim 1, it is characterised in that in terms of dry weight, the solution fiber clostridium
The inoculum concentration of (Clostridium cellulolyticum) ATCC35319 is 108~1010CFU/ grams of crude fibre raw material.
4. microbial flocculant as claimed in claim 1, it is characterised in that in step (2), the time of the anaerobic fermentation is
50~70 days.
5. the microbial flocculant as described in Claims 1 to 4 is any, it is characterised in that the slime bacteria includes：It is orange-yellow glutinous
Coccus (Myxococcus xanthus) CGMCC1.3865 and sorangium cellulosum (Soranguim cellulosum)
ATCC25532。
6. microbial flocculant as claimed in claim 5, it is characterised in that in terms of dry weight, the orange-yellow myxococcus
(Myxococcus xanthus) CGMCC1.3865's and sorangium cellulosum (Soranguim cellulosum) ATCC25532
Inoculum concentration is 109~1011CFU/ grams of crude fibre raw material.
7. microbial flocculant as claimed in claim 5, it is characterised in that in step (3), the temperature of the aerobic fermentation is
30~35 DEG C, fermentation time is 45~55 days.
8. application of the microbial flocculant in the rich phosphorus sewage of processing as described in claim 1~7 is any.
9. application as claimed in claim 8, it is characterised in that the dosage of the microbial flocculant is rich for 0~0.5g/L
Phosphorus sewage.
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| CN110923270A (en) * | 2019-12-06 | 2020-03-27 | 鹤山市东古调味食品有限公司 | Method for producing acetic acid by utilizing chemical ethanol fermentation |
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| CN103525870A (en) * | 2013-10-23 | 2014-01-22 | 嘉兴学院 | Microbial flocculant as well as preparation method and application thereof |
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2017
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| CN103525870A (en) * | 2013-10-23 | 2014-01-22 | 嘉兴学院 | Microbial flocculant as well as preparation method and application thereof |
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| CN110923270A (en) * | 2019-12-06 | 2020-03-27 | 鹤山市东古调味食品有限公司 | Method for producing acetic acid by utilizing chemical ethanol fermentation |
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