CN107937473A - A kind of microbial flocculant and its preparation method and application - Google Patents

A kind of microbial flocculant and its preparation method and application Download PDF

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CN107937473A
CN107937473A CN201711063441.6A CN201711063441A CN107937473A CN 107937473 A CN107937473 A CN 107937473A CN 201711063441 A CN201711063441 A CN 201711063441A CN 107937473 A CN107937473 A CN 107937473A
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microbial flocculant
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卢梅雅
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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
    • 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
    • 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
    • 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
    • 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

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Abstract

The invention discloses a kind of microbial flocculant and its preparation method and application.The preparation method includes：(1) flcos producing bacteria is inoculated with crude fibre raw material, carries out aerobic fermentation, obtains one grade fermemtation thing；(2) Clostridium baratii CGMCC1.208 is inoculated with one grade fermemtation product, carries out anaerobic fermentation, obtains second order fermentation thing；(3) after the water content of second order fermentation thing is down to 25~30%, slime bacteria is inoculated with second order fermentation thing, water content is made 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, obtain the microbial flocculant.The present invention carries out crude fibre raw material aerobic-anaerobic aerobic fermentation successively using flcos producing bacteria Clostridium baratii slime bacteria, and the flocculant of acquisition has excellent phosphor-removing effect to rich phosphorus sewage.

Description

A kind of microbial flocculant and its preparation method and application
Technical field
The present invention relates to technical field of sewage, and in particular to a kind of microbial flocculant and preparation method thereof and should With.
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 preparation method of microbial flocculant, can be obtained with excellent using the preparation method Phosphor-removing effect and microbial flocculant of low cost.
A kind of preparation method of microbial flocculant, including：
(1) flcos producing bacteria is inoculated with crude fibre raw material, carries out aerobic fermentation, obtains one grade fermemtation thing；
(2) Clostridium baratii (Clostridium pasteurianum) CGMCC1.208 is inoculated with one grade fermemtation product, Anaerobic fermentation is carried out, obtains second order fermentation thing；
(3) after the water content of the second order fermentation thing is down to 25~30%, 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：Mould (Mucor foenicola) CGMCC3.2212 of careless setation, long shoot trichoderma (Trichoderma Longibrachiatum) CGMCC3.5924, Tabin aspergillus (Aspergillus tubingensis) CGMCC3.3292, mountainous region Mould (Penicillium montanense) CGMCC 3.6588 and Rhizopus microsporus (Rhizopus microsporus) At least one of CGMCC3.3608.
It is of the invention that flcos producing bacteria is first inoculated with crude fibre raw material, the life of first time is carried out to crude fibre raw material using flcos producing bacteria Thing converts, and generation can make the metabolin of the calcium phosphate precipitation in sewage；But by one wheel aerobic fermentation after, flcos producing bacteria into Enter growth cycle end (decaying the phase), it is very limited that this allows for conversion of the flcos producing bacteria to crude fibre raw material.
Therefore, the present invention continues to be inoculated with Clostridium baratii (Clostridium pasteurianum) in one grade fermemtation thing CGMCC1.208 carries out anaerobic fermentation.The Clostridium baratii bacterial strain can not only utilize cellulose raw into the metabolism with flocculating function Thing, and in anaerobic fermentation process, between the metabolin produced during step (1) aerobic fermentation and anaerobic fermentation mistake The chemical reaction of complexity can all occur between the metabolin produced during the metabolin and aerobic fermentation that are produced in journey, so as to obtain There must be the flocculant of a variety of functional groups.
The present invention is further inoculated with slime bacteria in second order fermentation thing, and slime bacteria can be with remaining thick in second order fermentation thing Fibrous raw material is as source of nutrition, or can conciliate fiber clostridium with dead in second order fermentation thing or active relatively low flcos producing bacteria As source of nutrition, further 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 further includes：
②：Streptomyces parvus (Streptomyces parvus) CGMCC4.5927, streptomyces diastaticus slabstone color subspecies (Streptomyces diastaticus subsp.ardesiacus) CGMCC4.1682 and ash slightly red streptomyces At least one of (Streptomyces griseorubens) CGMCC4.5959.
The 2. organize flcos producing bacteria from the 1. group it is different, the 2. group be actinomyces (the 1. group be fungi), it is a discovery of the invention that above-mentioned Actinomyces can not only degraded cellulose carry out bioconversion to it, and antagonism is not present between fungi listed in the 1. group Act on, it is also possible to there are synergistic function between metabolins caused by both so that the dephosphorization efficiency higher of flocculant.
As further preferred, the flcos producing bacteria includes：Rhizopus microsporus (Rhizopus microsporus) CGMCC3.3608 and streptomyces parvus (Streptomyces parvus) CGMCC4.5927.
Before step (1) is inoculated with flcos producing bacteria, first crude fibre raw material can be crushed, degree of grinding is more thin better. Crude fibre utilization rate can not only be so improved, and also allows for flcos producing bacteria, Clostridium baratii and slime bacteria growth, subsequent extracted wadding During solidifying agent, higher extraction efficiency can be also obtained.
Crude fibre raw material can be crops, such as stalk, sweet potato dregs, bagasse etc..
After crushing, the water content of crude fibre raw material should be adjusted to 50~70%, pH and adjusted to 4~8, so as to be production wadding Bacterium provides the environment of suitable its growth breeding.
Preferably, in terms of dry weight, the 1. group and the inoculum concentration for 3. organizing flcos producing bacteria are 105~106A spore/gram thick fibre Tie up raw material；The 3. to organize the flcos producing bacteria speed of growth relatively slow, therefore its inoculum concentration should be higher than that the 1. group.
Preferably, in step (1), the temperature of the aerobic fermentation is 25~30 DEG C, and the aerobic fermentation time is 28~38 My god.
Preferably, in terms of dry weight, Clostridium baratii (Clostridium pasteurianum) CGMCC1.208's Inoculum concentration is 1010~1012CFU/ grams of crude fibre raw material.
Preferably, in step (2), the time of the anaerobic fermentation is 70~90 days.
Preferably, the slime bacteria is sorangium cellulosum (Soranguim cellulosum) ATCC25532.Fiber stack Cellulose remaining cellulose raw material as source of nutrition, can be carried out bioconversion, produced by capsule bacterium using in second order fermentation thing The macromolecular such as substantial amounts of mucus, the glycoprotein contained in mucus, mucopolysaccharide can be with the metabolin and reactant in second order fermentation thing Mixing, makes the flocculant of acquisition have more preferably flocculation ability and dephosphorization efficiency.
Preferably, in terms of dry weight, the inoculum concentration of the slime bacteria is 1010~1012CFU/ grams of crude fibre raw material.
Slime bacteria is inoculated after why the water content of second order fermentation thing being down to 25~30%, while after controlling inoculation The water content of three grade fermemtation matrix is below 40%, pH 6~8, be because slime bacteria is to the more demanding of own growth condition, Oxygen demand is high, if water content is excessive to be unfavorable for slime bacteria growth.
Preferably, in step (3), the temperature of the aerobic fermentation is 30~35 DEG C, and fermentation time is 70~90 days.It is fine It is longer to the fermentation period of cellulose raw material, it is necessary to which one section of longer time could farthest utilize cellulosic to tie up heap capsule bacterium Raw material.In the fermentation process of sorangium cellulosum, the metabolin of sorangium cellulosum can may also wad a quilt with cotton with having in second order fermentation thing Complicated chemical reaction occurs for solidifying and dephosphorization effect small molecule so that the flocculant finally obtained has more preferably flocculation ability And dephosphorization efficiency.
Present invention also offers the microbial flocculant obtained by the preparation method.Do not contained only in the microbial flocculant There are the metabolin that the generation of fiber clostridium is conciliate by flcos producing bacteria, and the reactant for the generation that reacts to each other between metabolin, also include Macromolecular, these macromoleculars and the metabolins and reactant in second order fermentation thing such as the glycoprotein of slime bacteria generation, mucopolysaccharide, Or mixing, or react to each other, further improve the flocculation ability and dephosphorization efficiency of flocculant.
Present invention also offers application of the microbial flocculant in rich phosphorus sewage disposal.
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 suitably. When the concentration of phosphate from sewage is in 160mg/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 is first inoculated with flcos producing bacteria in crude fibre raw material, and crude fibre raw material is carried out for the first time using flcos producing bacteria Bioconversion, generation can make the metabolin of calcium phosphate precipitation in sewage；And continue to be inoculated with Pasteur in one grade fermemtation thing Clostridium (Clostridium pasteurianum) CGMCC1.208 carries out anaerobic fermentation, which can not only profit With cellulose raw into the metabolin with flocculating function, and in anaerobic fermentation process, produced during step (1) aerobic fermentation Between the metabolin produced during the metabolin and aerobic fermentation that are produced between raw metabolin and in anaerobic fermentation process Complicated chemical reaction will occur, so as to obtain the flocculant with a variety of functional groups；
(2) 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 can make the metabolin of the calcium phosphate precipitation in sewage as source of nutrition, further generation, substantially increase raw material profit With 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；
Fig. 3 be middle flocculant in microbial flocculant prepared by comparative example 1 and comparative example 2 and embodiment 1 to containing The treatment effect of phosphorus simulated wastewater.
Embodiment
Technical scheme is described in further detail with reference to the accompanying drawings and detailed description.
Sorangium cellulosum (Soranguim cellulosum) ATCC25532 used in the present invention is trained from US mode Support what thing collection warehousing (American type culture collection) purchase obtained, remaining bacterial strain is from Chinese common micro- Biological inoculum preservation administrative center (China General Microbiological Culture Collection Center, CGMCC) purchase obtains.
Embodiment 1
A kind of preparation method of microbial flocculant of the present embodiment, including：
1st, activated spawn
(1) Rhizopus microsporus (Rhizopus microsporus) CGMCC3.3608 is inoculated in sterilized wheat bran culture In base, cultivated 5 days at 28 DEG C, treat the spore quantity of Rhizopus microsporus in culture more than 108During a/gram culture, obtain small Spore head mold microbial inoculum, 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) Clostridium baratii (Clostridium pasteurianum) CGMCC1.208 is inoculated in sterilized be equipped with In the 250mL anaerobism bottles of 100mL DCB-1 culture mediums (crushing maize stalk containing 5.0g/L), 35 DEG C are cultivated 3 days, collect bacterium Body, is made concentration as 1012The bacteria suspension of CFU/mL, it is stand-by.
(4) by sorangium cellulosum (Soranguim cellulosum) ATCC25532 (from American Type Culture collection warehousing (American type culture collection) purchase obtains) (training is inoculated in sterilized CNST solid mediums Support primary surface and lay the maize straw and bagasse 1 for having crushing:1 mixture), cultivated 7 days at 35 DEG C, collect gloeospore, system It is 10 into concentration15The 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 Rhizopus microsporus microbial inoculum 40g, above-mentioned small strepto- Bacterium 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 33 days, obtains one grade fermemtation thing；
(3) the one grade fermemtation thing in case is transferred in a plastic barrel, adds above-mentioned Clostridium baratii bacteria suspension 200mL；Pressure It is real, ensure to fill one grade fermemtation thing in plastic barrel, bung hole, which covers tightly, to be shut, and plastic barrel is placed in progress constant-temperatureanaerobic anaerobic hair at 32 DEG C Ferment, obtains second order fermentation product (water content 25%) after 80 days；
(4) pH of second order fermentation product is adjusted to after 7~7.5, the gloeospore for adding the above-mentioned sorangium cellulosums of 200mL hangs Liquid, stirs evenly, and is transferred to four walls and is equipped with the plastic box of air hole, and it is good that plastic box is placed in progress constant temperature in 32 DEG C Aerobe fermentation, obtains three grade fermemtation thing after 80 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.
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 preparation method of microbial flocculant of the present embodiment, including：
1st, activated spawn
(1) Tabin aspergillus (Aspergillus tubingensis) CGMCC3.3292 is inoculated in sterilized wheat bran to train Support in base, cultivated 5 days at 28 DEG C, treat the spore quantity of Tabin aspergillus in culture more than 108During a/gram culture, obtain Tabin aspergillus microbial inoculum, it is stand-by.
(2) by streptomyces diastaticus slabstone color subspecies (Streptomyces diastaticus Subsp.ardesiacus) CGMCC4.1682 is inoculated in sterilized Gause I slant medium, and 7 are cultivated at 30 DEG C After it, the sterile water with bead is injected into test tube slant, fully after vibration, monospore suspension is filtered into, using photoelectricity ratio Turbid method detects spore count, and spore quantity is made as 109The spore suspension of a/mL, it is stand-by.
(3) Clostridium baratii (Clostridium pasteurianum) CGMCC1.208 is inoculated in sterilized be equipped with In the 250mL anaerobism bottles of 100mL DCB-1 culture mediums (crushing maize stalk containing 5.0g/L), 35 DEG C are cultivated 3 days, collect bacterium Body, is made concentration as 1012The bacteria suspension of CFU/mL, it is stand-by.
(4) 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 1014The 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 Tabin aspergillus microbial inoculum 35g, above-mentioned amylase Streptomycete slabstone color subspecies spore suspension 110mL, 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 Clostridium baratii bacteria suspension 180mL；Pressure It is real, ensure to fill one grade fermemtation thing in plastic barrel, bung hole, which covers tightly, to be shut, and plastic barrel is placed in progress constant-temperatureanaerobic anaerobic hair at 32 DEG C Ferment, obtains second order fermentation product (water content 20%) after 70 days；
(4) pH of second order fermentation product is adjusted to after 7~7.5, the gloeospore for adding the above-mentioned sorangium cellulosums of 180mL hangs Liquid, stirs evenly, and is transferred to four walls and is equipped with the plastic box of air hole, and it is good that plastic box is placed in progress constant temperature in 32 DEG C Aerobe fermentation, obtains three grade fermemtation thing after 70 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 preparation method of microbial flocculant of the present embodiment, including：
1st, activated spawn
(1) mountainous region mould (Penicillium montanense) CGMCC 3.6588 is inoculated in sterilized wheat bran to train Support in base, cultivated 5 days at 28 DEG C, treat the spore quantity of mountainous region mould in culture more than 108During a/gram culture, obtain Mountainous region mould microbial inoculum, it is stand-by.
(2) ash slightly red streptomyces (Streptomyces griseorubens) CGMCC4.5959 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) Clostridium baratii (Clostridium pasteurianum) CGMCC1.208 is inoculated in sterilized be equipped with In the 250mL anaerobism bottles of 100mL DCB-1 culture mediums (crushing maize stalk containing 5.0g/L), 35 DEG C are cultivated 3 days, collect bacterium Body, is made concentration as 1012The bacteria suspension of CFU/mL, it is stand-by.
(4) 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 1015The 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%；Above-mentioned mountainous region mould microbial inoculum 65g is added, above-mentioned ash is slightly red Streptomycete spore suspension 160mL, 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 38 days, obtains one grade fermemtation thing；
(3) the one grade fermemtation thing in case is transferred in a plastic barrel, adds above-mentioned Clostridium baratii bacteria suspension 360mL；Pressure It is real, ensure to fill one grade fermemtation thing in plastic barrel, bung hole, which covers tightly, to be shut, and plastic barrel is placed in progress constant-temperatureanaerobic anaerobic hair at 32 DEG C Ferment, obtains second order fermentation product (water content 30%) after 90 days；
(4) pH of second order fermentation product is adjusted to after 7~7.5, the gloeospore for adding the above-mentioned sorangium cellulosums of 450mL hangs Liquid, stirs evenly, and is transferred to four walls and is equipped with the plastic box of air hole, and it is good that plastic box is placed in progress constant temperature in 32 DEG C Aerobe fermentation, obtains three grade fermemtation thing after 90 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.
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, 160mg/L), 0.01g is put into respectively into first group of each beaker Dephosphorization microbial flocculant prepared by embodiment 1, removing for the preparation of 0.01g embodiments 2 is put into second group of each beaker respectively Phosphorus microbial flocculant, the dephosphorization microbial flocculant of the preparation of 0.01g embodiments 3 is put into the 3rd group of each beaker respectively, Stir evenly, in 100r/min, vibrate mixing 40min at room temperature, centrifugation, goes to precipitate, and takes supernatant, detects phosphoric acid in supernatant The concentration of potassium dihydrogen, calculates the dephosphorization microbial flocculant of each embodiment to the removal efficiency of potassium dihydrogen phosphate, the result is shown in Figure 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 92% treatment effeciency, processing of the dephosphorization microbial flocculant that wherein prepared by embodiment 1 to phosphorous simulated wastewater Effect is best, still has 99.6% dephosphorization efficiency when phosphorus concentration is 87.3mg/L in phosphorous simulated wastewater, and phosphorous simulation is useless Still there is 67.1% dephosphorization efficiency when phosphorus concentration is 160mg/L in water.
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 160mg/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, stir evenly, and in 100r/min, vibrate mixing 40min at room temperature, and centrifugation, goes to precipitate, Supernatant is taken, detects the concentration of potassium dihydrogen phosphate in supernatant, calculates each group dephosphorization microbial flocculant to potassium dihydrogen phosphate Removal efficiency, 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 84.8%, 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 0.002mg/L in two parts of river waters, treatment effeciency reaches 96.7%.
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 Rhizopus microsporus microbial inoculum.
It is prepared by flocculant and two level centre flocculant among the level-one in Example 1, and comparative example 1 and comparative example 2 Microbial flocculant；12, beaker is taken again, is divided into four groups, pours into the phosphorus that 100mL concentration is 5mg/L respectively in every group of beaker Acid dihydride potassium solution, flocculant among 0.01g level-ones is put into first group of each beaker, is put into second group of each beaker Flocculant among 0.01g two levels, 1 microbial flocculant of 0.01g comparative examples is put into the 3rd group of each beaker, to the 4th group 2 microbial flocculant of 0.01g comparative examples is put into each beaker；Stir evenly, in 100r/min, vibrate mixing 40min at room temperature, Centrifugation, goes to precipitate, and takes supernatant, detects the concentration of potassium dihydrogen phosphate in supernatant, calculates each flocculant to potassium dihydrogen phosphate Removal efficiency, the result is shown in Fig. 3.
As seen from Figure 3, the anaerobic fermentation that Clostridium baratii carries out can improve the dephosphorization efficiency of flocculant among level-one, and fine The aerobic fermentation that dimension heap capsule bacterium carries out can further improve the dephosphorization efficiency of flocculant among two level again.And streptomyces parvus or small spore The dephosphorization for the flocculant that the dephosphorization efficiency of the flocculant obtained during head mold individualism is obtained without both in the presence of at the same time is imitated Rate is high, and indicating this, there may be there may be synergy between the metabolin of collaboration symbiosis, or both between the two.

Claims (10)

  1. A kind of 1. preparation method of microbial flocculant, it is characterised in that including：
    (1) flcos producing bacteria is inoculated with crude fibre raw material, carries out aerobic fermentation, obtains one grade fermemtation thing；
    (2) Clostridium baratii (Clostridium pasteurianum) CGMCC1.208 is inoculated with one grade fermemtation product, is carried out Anaerobic fermentation, obtains second order fermentation thing；
    (3) after the water content of the second order fermentation thing is down to 25~30%, 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：Mould (Mucor foenicola) CGMCC3.2212 of careless setation, long shoot trichoderma (Trichoderma Longibrachiatum) CGMCC3.5924, Tabin aspergillus (Aspergillus tubingensis) CGMCC3.3292, mountainous region Mould (Penicillium montanense) CGMCC 3.6588 and Rhizopus microsporus (Rhizopus microsporus) At least one of CGMCC3.3608.
  2. 2. the preparation method of microbial flocculant as claimed in claim 1, it is characterised in that the flcos producing bacteria further includes：
    ②：Streptomyces parvus (Streptomyces parvus) CGMCC4.5927, streptomyces diastaticus slabstone color subspecies (Streptomyces diastaticus subsp.ardesiacus) CGMCC4.1682 and ash slightly red streptomyces At least one of (Streptomyces griseorubens) CGMCC4.5959.
  3. 3. the preparation method of microbial flocculant as claimed in claim 2, it is characterised in that the flcos producing bacteria includes：Small spore Head mold (Rhizopus microsporus) CGMCC3.3608, Clostridium baratii (Clostridium pasteurianum) CGMCC1.208 and streptomyces parvus (Streptomyces parvus) CGMCC4.5927.
  4. 4. the preparation method of the microbial flocculant as described in claims 1 to 3 is any, it is characterised in that described in terms of dry weight The inoculum concentration of Clostridium baratii (Clostridium pasteurianum) CGMCC1.208 is 1010~1012CFU/ grams of crude fibre original Material.
  5. 5. the preparation method of the microbial flocculant as described in claims 1 to 3 is any, it is characterised in that in step (2), institute The time for stating anaerobic fermentation is 70~90 days.
  6. 6. the preparation method of the microbial flocculant as described in claims 1 to 3 is any, it is characterised in that the slime bacteria is Sorangium cellulosum (Soranguim cellulosum) ATCC25532.
  7. 7. the preparation method of the microbial flocculant as described in claim 1 or 6, it is characterised in that described viscous thin in terms of dry weight The inoculum concentration of bacterium is 1010~1012CFU/ grams of crude fibre raw material.
  8. 8. the preparation method of microbial flocculant as claimed in claim 1, it is characterised in that in step (3), the aerobic hair The temperature of ferment is 30~35 DEG C, and fermentation time is 80~100 days.
  9. 9. the microbial flocculant obtained by any preparation method of claim 1~8.
  10. 10. application of the microbial flocculant as claimed in claim 9 in rich phosphorus sewage disposal.
CN201711063441.6A 2017-10-26 2017-10-26 A kind of microbial flocculant and its preparation method and application Withdrawn CN107937473A (en)

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