CN106636058A - Preparation method of nanometer material/group sensing quenching enzyme particle modified composite membrane - Google Patents
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- C12Y305/01—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in linear amides (3.5.1)
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Abstract
The invention discloses a preparation method of a nanometer material/group sensing quenching enzyme particle modified composite membrane. According to the preparation method of the nanometer material/group sensing quenching enzyme particle modified composite film, oxidized graphene, graphene or carbon nanotubes are used as carriers; the static electricity adsorption effect is used for immobilizing bacterium groups to sense quenching enzymes; the prepared loaded particle ensures the quenching enzyme activity to the maximum degree. Meanwhile, the particle is added into high molecular polymer membrane forming liquid; a phase conversion method is used for preparing the nanometer material/group sensing quenching enzyme modified composite membrane. The composite membrane has the advantages that the functions of the bacterium signal molecules can be degraded by the quenching enzymes; the growth of the membrane surface biological membrane can be inhibited; meanwhile, the organic composite membrane has the effects of easy operation of the preparation method and good biological membrane pollution resistant effect, and can be widely applied to all fields such as biology, medicine, food and water treatment.
Description
Technical field
The invention belongs to application of membrane separation technology field, is related to a kind of nano material/Quorum quenching enzymes granule modified
Compound membrane preparation method.
Background technology
In current water-treatment technology field, membrane separation technique is still one of mainstream technology, and separation membrane has separation
The features such as effect is good, feature is strong, is obtained in every field such as water process, biology, medicine and widely applies.But in film
During materials'use, the easily phenomenon such as pollution, susceptible to plugging is there is, affect the service life of membrane material.In control fouling membrane
Method in, including membrane material itself modified, the modified bag of the various methods such as physics, Chemical cleaning, wherein membrane material itself
Including improves the hydrophilicity of membrane material, and antibacterial nano-Ag particles, lysozyme etc. are added in membrane material.
In recent years, scientists study finds that when the quantity accumulation of bacterium increases to a certain degree, bacterium can lead to
Signaling molecule signalling is crossed, changes and coordinate the behavior between them, this regulatory mechanism is referred to as the quorum sensing of bacterium
(quorum sensing,QS).The quorum sensing of bacterium can regulate and control the formation of bacterial biof iotalm, and bioluminescence, pigment is produced,
Antibiotic is formed, plasmid Conjugative tiansfer etc..In recent years scientists found that quorum sensing inhibitor can be with interference signal molecule system
The conduction of system, suppresses the expression of certain characteristic of bacterium and mechanism.Researchers' trial will be quenched enzyme and be introduced directly in membrane material, i.e.,
The addition that enzyme and nano material can be passed through changes the structure of membrane material, while and can reach the purpose for suppressing biofilm development.
But, when enzyme being quenched being used alone, can existence and stability it is bad, the shortcomings of easy in inactivation.Therefore, typically can select
Enzyme is made composite and is applied by suitable fixed material.Carbon nanomaterial is study hotspot in recent years, is mainly included
The materials such as graphene oxide, Graphene, CNT, carbon fiber.Carbon nanomaterial has nano-grade size, nanoscale due to it
Lamellar structure and the well characteristic such as absorption, are used in many fields, and have obtained quickly widely promoting.Using nanometer material
The powerful specific surface of material and electrostatic adsorption immobilized enzyme, can improve the fixed amount of enzyme can not cause big to the activity of enzyme again
Destruction, becomes a kind of preferable enzyme immobilization carrier.
The content of the invention
The present invention provides a kind of nano material/Quorum quenching enzymes granule modified compound membrane preparation method.
The purpose of the present invention is realized by following technical proposals.
A kind of granule modified compound membrane preparation method of nano material/Quorum quenching enzymes, comprises the steps:
1) preparation of nano material/Quorum quenching enzymes particle：By 25-50mg include graphene oxide, Graphene and
The nano material of CNT is dispersed in 100-200mL PBSs, room temperature 30～60min of ultrasound, obtains nanometer
Material mixed solution；75-150mg is quenched into enzyme to be dispersed in nano material mixed solution, room temperature 30～60min of ultrasound is obtained
Nano material/Quorum quenching enzymes particle mixed solution, vacuum freeze drying obtains final product nano material/Quorum quenching enzymes
Particle, freezen protective is standby；
2) preparation of composite membrane casting solution：It is 0.1～1% nano material/Quorum quenching enzymes particle by mass fraction
In adding 80～85% organic solvents, 15～30min of ultrasound；0.1～1% polymeric additive of addition, high-speed stirred 30min,
14～26% high molecular polymers, high-temperature stirring are added to be completely dissolved to high molecular polymer, standing and defoaming；
3) film forming solidification：By step 2) casting solution for preparing is placed on the glass plate of smooth clean, and put down with scraper
Paving on a glass, is immersed in film forming in deionized water glass plate level rapidly；The film for making is put in deionized water extremely
Few 24h, obtains the granule modified composite membrane of nano material/Quorum quenching enzymes.
Further, the preparation method of the Graphene is mechanical stripping method, vapour deposition process (CVD), pyrolysis SiC methods, oxygen
Graphite alkene reducing process, carbon pipe patterning method and liquid phase less than the three layers Graphenes that directly prepared by ultrasound stripping method.
Further, the CNT includes single wall, double-walled and multi-walled carbon nano-tubes, and caliber is 1～100nm, and pipe range is
0.5～100 μm.
Further, the PBS is in mass ratio (0.68)：(0.12):Potassium dihydrogen phosphate, the chlorine of 1 configuration
Change the mixed solution of sodium and ultra-pure water configuration.Further, the step 1) in, 1～1.5h is stirred at 0～10 DEG C.
Further, the enzyme that is quenched is including all to bacterium signal including amino-acylase, lactonase or decarboxylase
What molecule had a degradation is quenched enzyme.
Further, the high molecular polymer can be by other polymers polypropylene nitrile, Kynoar, polysulfones, poly-
One kind in ether sulfone, polyurethane and cellulose acetate class.
Further, the organic solvent includes N-N dimethyl acetamides, N-N dimethylformamides, dimethyl sulfoxide or N- first
One or more of base pyrrolidones.
Further, the polymeric additive selected from polyvinylpyrrolidone, polyvinyl alcohol and polyethylene glycol, lithium chloride,
One or more in acrylic acid or beta-schardinger dextrin.
Further, the granule modified composite membrane deionized water of the nano material/Quorum quenching enzymes is rinsed, is put into
4 DEG C of preservations in deionized water.
The beneficial effects of the present invention is：
(1) preparation method of the granule modified composite membrane of nano material/Quorum quenching enzymes, composite membrane is except dropping
Solution Bacterial signal molecules, again may be by hydrolyzing the amino acids materials such as amino, degraded methionine.
(2) improve and be quenched that enzyme dead load amount efficiency is low, diffusion restriction effect, activity of the immobilized enzyme be not high, and with
When traditional chemical bond is closed, the problems such as enzymatic activity is restricted and affects enzyme activity.
(3) low-dimensional carbon nanomaterial easily prepare, it is easy to maintain, specific surface area is big, and big destruction is not resulted in enzyme activity.
(4) preparation method of the granule modified composite membrane of nano material/Quorum quenching enzymes is simple, can it is biological,
The multiple fields such as medical science, environment are applied.
Specific embodiment
The present invention is described in further details with reference to specific embodiment, but is not intended as doing any restriction to invention
Foundation.
A kind of granule modified compound membrane preparation method of nano material/Quorum quenching enzymes, comprises the steps:
1) preparation of nano material/Quorum quenching enzymes particle：By 25-50mg include graphene oxide, Graphene and
The nano material of CNT is dispersed in 100-200mL PBSs, room temperature 30～60min of ultrasound, obtains nanometer
Material mixed solution；75-150mg is quenched into enzyme to be dispersed in nano material mixed solution, room temperature 30～60min of ultrasound is obtained
Nano material/Quorum quenching enzymes particle mixed solution, vacuum freeze drying obtains final product nano material/Quorum quenching enzymes
Particle, freezen protective is standby；
The preparation method of the Graphene is mechanical stripping method, vapour deposition process (CVD), pyrolysis SiC methods, graphene oxide
Reducing process, carbon pipe patterning method and liquid phase less than the three layers Graphenes that directly prepared by ultrasound stripping method.CNT includes single wall, double
Wall and multi-walled carbon nano-tubes, caliber is 1～100nm, and pipe range is 0.5～100 μm.
The PBS is in mass ratio (0.68)：(0.12):1 configuration potassium dihydrogen phosphate, sodium chloride and
The mixed solution of ultra-pure water configuration.It is including all to bacterium letter including amino-acylase, lactonase or decarboxylase that enzyme is quenched
What number molecule had a degradation is quenched enzyme.
2) preparation of composite hyperfiltration membrane casting solution：It is 0.1～1% nano material/Quorum quenching enzymes by mass fraction
Particle adds 80～85% organic solvents (N-N dimethyl acetamides, N-N dimethylformamides, dimethyl sulfoxide or N- methylpyrroles
In one or more of alkanone), 15～30min of ultrasound；Add 0.1～1% polymeric additive (polyvinylpyrrolidone, poly-
In vinyl alcohol and polyethylene glycol, lithium chloride, acrylic acid or beta-schardinger dextrin one or more), high-speed stirred 30min, add 14
～26% high molecular polymer (in polyacrylonitrile, Kynoar, polysulfones, polyether sulfone, polyurethane and cellulose acetate class
Kind), high-temperature stirring is completely dissolved to high molecular polymer, standing and defoaming；
3) film forming solidification：By step 2) casting solution for preparing is placed on the glass plate of smooth clean, and put down with scraper
Paving on a glass, is immersed in film forming in deionized water glass plate level rapidly；The film for making is put in deionized water extremely
Few 24h, obtains the granule modified composite membrane of nano material/Quorum quenching enzymes, and then deionized water is rinsed, and is put into deionization
4 DEG C of preservations in water.
The present invention is illustrated below by specific embodiment.
Embodiment 1：
The preparation of graphene oxide uses Hummer improved methods.Natural flake graphite powder used is 100 purposes
Powder, purity is 99.999＆.
1) preparation of nano material/Quorum quenching enzymes particle：50mg graphene oxides are dispersed in into 150mL0.02M
In the PBS of pH (6.9), room temperature ultrasound 30min adds 150mg amino-acylases, and at 0 DEG C 1h is stirred, and surpasses
Sound 30min, vacuum freeze drying obtains final product graphene oxide/amino-acylase hybrid particulates, and freezen protective is standby.
2) preparation of composite membrane casting solution：0.1wt% graphene oxides/amino-acylase hybrid particulates are added
84.9wt%N, in N-dimethylformamide, ultrasonic 15min；1wt% polyvinyl alcohol, high-speed stirred 30min is added to add
14wt% Kynoar, 40 DEG C of stirrings, is completely dissolved, standing and defoaming to Kynoar.
3) film forming solidification：By step 2) casting solution for preparing is placed on the glass plate of smooth clean, and put down with scraper
Paving on a glass, is immersed in film forming in 10 DEG C of deionized waters glass plate level rapidly.The film for making is placed on into deionized water
Middle 24h.Obtain the granule modified composite hyperfiltration membrane of nano material/Quorum quenching enzymes.Film deionized water is rinsed, is put into
4 DEG C of preservations in ionized water.
Embodiment 2：
The preparation of Graphene adopts hydrazine hydrate reduction graphene oxide method：By the graphene oxide (100mg) in embodiment 1
In being distributed to the aqueous solution (100ml), ultrasonic disperse 50min is subsequently adding hydrazine hydrate (2ml, 50%w/v), returns after being well mixed
24h is reacted under the conditions of stream.After reaction terminates, sample filtering separating, washing, and dry in 60 DEG C of vacuum drying oven.
1) preparation of nano material/Quorum quenching enzymes particle：By 25mg graphene dispersions in 100mL0.2M pH
(7.0) in PBS, room temperature ultrasound 40min adds 100mg amino-acylases, and 1h is stirred at 4 DEG C, ultrasound
40min, vacuum freeze drying obtains final product Graphene/amino-acylase hybrid particulates, and freezen protective is standby.
2) preparation of composite membrane casting solution：0.3wt% Graphenes/amino-acylase hybrid particulates are added into 80wt% diformazans
Sulfoxide and N methyl pyrrolidones are according to 1:In 1 mixed liquor, ultrasonic 30min；0.5wt% polyethylene glycol and lithium chloride is added to press
According to 1:1 ratio mixing, high-speed stirred 30min adds 19.2wt% polysulfones, 45 DEG C of stirrings to be completely dissolved to Kynoar,
Standing and defoaming.
3) film forming solidification：By step 2) casting solution for preparing is placed on the glass plate of smooth clean, and put down with scraper
Paving on a glass, is immersed in film forming in 10 DEG C of deionized waters glass plate level rapidly.The film for making is placed on into deionized water
Middle 24h.Obtain the granule modified composite hyperfiltration membrane of nano material/Quorum quenching enzymes.Film deionized water is rinsed, is put into
4 DEG C of preservations in ionized water.
Embodiment 3：
1) preparation of nano material/Quorum quenching enzymes particle：30mg CNTs are dispersed in into 200mL0.2M pH
(7.0) in PBS, room temperature ultrasound 60min adds 75mg amino-acylases, and 1h is stirred at 10 DEG C, ultrasound
60min, vacuum freeze drying obtains final product CNT/amino-acylase hybrid particulates, and freezen protective is standby.
2) preparation of composite membrane casting solution：1wt% CNTs/amino-acylase hybrid particulates are added into 72.9wt%N,
In N dimethyl acetamide, ultrasonic 20min；0.1wt% polyvinylpyrrolidones and beta-schardinger dextrin are added according to 1:1 ratio is mixed
Close, high-speed stirred 30min, add 26wt% cellulose acetates, 40 DEG C of stirrings to be completely dissolved to cellulose acetate, standing and defoaming.
3) film forming solidification：By step 2) casting solution for preparing is placed on the glass plate of smooth clean, and put down with scraper
Paving on a glass, is immersed in film forming in 10 DEG C of deionized waters glass plate level rapidly.The film for making is placed on into deionized water
Middle 24h.Obtain the granule modified composite hyperfiltration membrane of nano material/Quorum quenching enzymes.Film deionized water is rinsed, is put into
4 DEG C of preservations in ionized water.
The water flux of three of the above milipore filter and enzyme membrane activity number of days see attached list 1.
Subordinate list 1
As it can be seen from table 1 the present invention prepares the granule modified composite membrane of nano material/Quorum quenching enzymes, its
2000ppm NaCl solution water fluxes are not less than 26L/ (M2H), BSA retentions are not less than 78%；Enzyme membrane activity number of days is 28d.
As can be seen from Table 1, the present invention prepared by the granule modified composite membrane of nano material/Quorum quenching enzymes not only
With higher separating property, excellent bioactive properties are provided simultaneously with, are a kind of good organic hybrid films of processing characteristics.
The above, be only the present invention for a kind of granule modified composite membrane of nano material/Quorum quenching enzymes and its
The embodiment of preparation method, not does any restriction to the present invention, it is every according to the technology of the present invention to made for any of the above embodiments
Any simple modification, change and equivalent structure change, still fall within the protection domain of technical solution of the present invention.
Claims (10)
1. the granule modified compound membrane preparation method of a kind of nano material/Quorum quenching enzymes, it is characterised in that including following steps
Suddenly:
1) preparation of nano material/Quorum quenching enzymes particle：25-50mg is received including graphene oxide, Graphene and carbon
The nano material of mitron is dispersed in 100-200mL PBSs, room temperature 30～60min of ultrasound, obtains nano material
Mixed solution；75-150mg is quenched into enzyme to be dispersed in nano material mixed solution, room temperature 30～60min of ultrasound obtains nanometer
Material/Quorum quenching enzymes particle mixed solution, vacuum freeze drying obtains final product nano material/Quorum quenching enzymes particle,
Freezen protective is standby；
2) preparation of composite hyperfiltration membrane casting solution：It is 0.1～1% graphite alkenyl nanometer materials/bacterial flora body-sensing by mass fraction
Enzyme hybrid particulates should be quenched to add in 80～85% organic solvents, 15～30min of ultrasound；Add the addition of 0.1～1% macromolecule
Agent, high-speed stirred 30min adds 14～26% high molecular polymers, and high-temperature stirring is completely dissolved to high molecular polymer, quiet
Put deaeration；
3) film forming solidification：By step 2) casting solution for preparing is placed on the glass plate of smooth clean, and is laid in scraper
On glass plate, rapidly glass plate level is immersed in film forming in deionized water；The film for making is put in deionized water at least
24h, obtains the granule modified composite membrane of nano material/Quorum quenching enzymes.
2. method according to claim 1, it is characterised in that the preparation method of the Graphene is mechanical stripping method, gas
The three of the direct ultrasound stripping method preparation of phase sedimentation CVD, pyrolysis SiC methods, graphene oxide reducing process, carbon pipe patterning method and liquid phase
The following Graphene of layer.
3. method according to claim 1, it is characterised in that the CNT includes single wall, double-walled and many wall carbon are received
Mitron, caliber is 1～100nm, and pipe range is 0.5～100 μm.
4. method according to claim 1, it is characterised in that the PBS is in mass ratio (0.68)：
(0.12):The mixed solution of the potassium dihydrogen phosphate, sodium chloride and ultra-pure water configuration of 1 configuration.
5. method according to claim 1, it is characterised in that the step 1) in, 1～1.5h is stirred at 0～10 DEG C.
6. method according to claim 1, it is characterised in that it is described be quenched enzyme be include amino-acylase, lactonase or
Decarboxylase it is interior it is all there is degradation to Bacterial signal molecules enzyme is quenched.
7. method according to claim 1, it is characterised in that the high molecular polymer includes polyacrylonitrile, gathers inclined fluorine
One kind in ethene, polysulfones, polyether sulfone, polyurethane and cellulose acetate class.
8. method according to claim 1, it is characterised in that the organic solvent includes N-N dimethyl acetamides, N-N
One or more of dimethylformamide, dimethyl sulfoxide or 1-METHYLPYRROLIDONE.
9. method according to claim 1, it is characterised in that the polymeric additive selected from polyvinylpyrrolidone,
One or more in polyvinyl alcohol and polyethylene glycol, lithium chloride, acrylic acid or beta-schardinger dextrin.
10. method according to claim 1, it is characterised in that change the nano material/Quorum quenching enzymes particle
Property composite membrane preparation method deionized water rinse, be put in deionized water 4 DEG C of preservations.
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| CN108949740A (en) * | 2018-08-24 | 2018-12-07 | 西安建筑科技大学 | A kind of preparation method of nano material/Quorum quenching enzymes hydrogel |
| CN109046046A (en) * | 2018-08-24 | 2018-12-21 | 西安建筑科技大学 | A kind of preparation method of nano material/Quorum quenching enzymes hydrogel nanofiltration membrane |
| CN109134950A (en) * | 2018-08-24 | 2019-01-04 | 西安建筑科技大学 | A kind of preparation method embedding quorum sensing inhibitor hydrogel blend film |
| CN109180982A (en) * | 2018-08-24 | 2019-01-11 | 西安建筑科技大学 | A kind of preparation method of embedded nano material/Quorum quenching enzymes composite pellets |
| CN109761348A (en) * | 2019-02-28 | 2019-05-17 | 北京工业大学 | A method of improving the anti-matrix impact capacity of anaerobic ammonium oxidation granular |
| CN111068529A (en) * | 2019-12-26 | 2020-04-28 | 沃克福德环境科技(深圳)有限责任公司 | Method for preparing anti-pollution membrane by using silicon dioxide nano material |
| US11441143B2 (en) * | 2019-01-31 | 2022-09-13 | Korea Univercity Research and Business Foundation | Enzyme-carrier complex |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949740A (en) * | 2018-08-24 | 2018-12-07 | 西安建筑科技大学 | A kind of preparation method of nano material/Quorum quenching enzymes hydrogel |
| CN109046046A (en) * | 2018-08-24 | 2018-12-21 | 西安建筑科技大学 | A kind of preparation method of nano material/Quorum quenching enzymes hydrogel nanofiltration membrane |
| CN109134950A (en) * | 2018-08-24 | 2019-01-04 | 西安建筑科技大学 | A kind of preparation method embedding quorum sensing inhibitor hydrogel blend film |
| CN109180982A (en) * | 2018-08-24 | 2019-01-11 | 西安建筑科技大学 | A kind of preparation method of embedded nano material/Quorum quenching enzymes composite pellets |
| US11441143B2 (en) * | 2019-01-31 | 2022-09-13 | Korea Univercity Research and Business Foundation | Enzyme-carrier complex |
| CN109761348A (en) * | 2019-02-28 | 2019-05-17 | 北京工业大学 | A method of improving the anti-matrix impact capacity of anaerobic ammonium oxidation granular |
| CN111068529A (en) * | 2019-12-26 | 2020-04-28 | 沃克福德环境科技(深圳)有限责任公司 | Method for preparing anti-pollution membrane by using silicon dioxide nano material |
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| CN106636058B (en) | 2019-07-12 |
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