CN105798324B - One kind is based on self-assembled structures analogue enztme and preparation method and application - Google Patents
One kind is based on self-assembled structures analogue enztme and preparation method and application Download PDFInfo
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- CN105798324B CN105798324B CN201610165756.0A CN201610165756A CN105798324B CN 105798324 B CN105798324 B CN 105798324B CN 201610165756 A CN201610165756 A CN 201610165756A CN 105798324 B CN105798324 B CN 105798324B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F9/00—Making metallic powder or suspensions thereof
- B22F9/16—Making metallic powder or suspensions thereof using chemical processes
- B22F9/18—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds
- B22F9/24—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds starting from liquid metal compounds, e.g. solutions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F1/00—Metallic powder; Treatment of metallic powder, e.g. to facilitate working or to improve properties
- B22F1/05—Metallic powder characterised by the size or surface area of the particles
- B22F1/054—Nanosized particles
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
Abstract
The invention discloses a kind of preparation method based on self-assembled structures analogue enztme, comprise the following steps：(1) chemical dissolution of self assembly behavior will can occur in water；(2) to adding active metal salt solution, reducing agent in above-mentioned (1), stirring, produce and described be based on self-assembled structures analogue enztme.In preparation method provided by the invention, two kinds of chemicals aqueous solution that tissue can occur prepared in advance are mixed, two kinds of molecules are fitted together by weak intermolecular force, ionizable metal salt is in its adsorption, in the presence of reducing agent, in-situ preparation metal nanoparticle, it is resulting that there is good peroxidase activity based on the metal nanoparticle of self-assembled structures under the conditions of wide pH and temperature range, high ionic strength etc..The materials synthesis mild condition is easy, low in raw material price, is easily commercially exploited.The metal nanoparticle material based on self-assembled structures of the present invention can also be applied to the numerous areas such as electro-catalysis, water process, biomedicine, have wide practical use.
Description
Technical field
The present invention relates to high-molecular compound field, specifically discloses one kind and is based on self-assembled structures analogue enztme and its system
Preparation Method and application.
Background technology
Compared with protease, although nano material analogue enztme has acid, alkali and high-temperature stability well, cost is cheap,
Synthesis is simple, can largely prepare, be easy to by types of functionality molecular modification and mark, but still suffers from very big gap, and what is faced chooses
War is still very severe.Common nano material analogue enztme Fe3O4, CeO2, nanogold analog derivative, graphite vinyl derivative etc. are all
It can be used to combine specific oxidizing ferment (such as glucose oxidase) and developer (such as 3,3', 5,5'- tetramethyl benzidine
TMB etc.) carry out specific molecular detection.It is difficult to be commercially exploited applied to people although achieving certain Detection results
Daily life in, its main cause has：A) the common high catalytic activity condition of easily prepared nano material analogue enztme is in acid
Property environment, almost without catalytic effect under neutrallty condition, and native enzyme (such as glucose oxidase) have in neutral conditions it is good
Good catalytic effect, protease may inactivate under acid condition, it is difficult to combine exploitation and examination that protease is applied to kit
The development of paper.Claim to prepare the nano material analogue enztme that can play high catalytic activity in neutral conditions although having been reported,
Its harsh preparation condition is difficult to be commercially employed；B) catalytic reaction activity is low, although some nano material analogue enztmes can be used for
Enzyme linked immunological marks, but its catalytic activity still can not be compared with horseradish peroxidase, it is difficult to be applied to enzyme linked immunological examination
The exploitation of agent box.
The content of the invention
The problem of in order to overcome prior art to exist, the present invention provide one kind and are based on self-assembled structures analogue enztme and its preparation
Method and application.
Present invention firstly provides a kind of preparation method based on self-assembled structures analogue enztme, comprise the following steps：
(1) compound that self assembly behavior can occur is dissolved in the water；
(2) to adding active metal salt solution, reducing agent in above-mentioned (1), stirring, produce and described be based on self-assembled structures mould
Intend enzyme.
Further, the chemical substance that self assembly behavior can occur is at least one of following substances combination：Trimerization
Cyanamide and cyanuric acid, melamine and ellagic acid, melamine and catechol -3,5- disulfonic acid, melamine and lemon
Acid, melamine and trimesic acid, melamine and trithiocyanuric acid, cyanuric acid and citric acid, cyanuric acid and equal benzene three
Acid.
Further, the active metal salt is at least one of following substances：Copper sulphate, copper chloride, copper acetate, nitric acid
Copper, silver nitrate, gold chloride, platinum chloride, palladium bichloride, palladium nitrate, iron chloride, ferric nitrate, frerrous chloride, ferric sulfate, cobalt chloride, nitre
Sour cobalt, nickel nitrate, nickel chloride.
Further, the reducing agent is at least one of following substances：Sodium borohydride, potassium borohydride, ascorbic acid, lemon
Lemon acid sodium, citric acid, ellagic acid, catechol -3,5- disulfonic acid.
Further, the concentration of the chemicals that self assembly behavior can occur can be 0.01~10mmol/L, specifically
Can be 0.01mmol/L, 0.05mmol/L, 0.1mmol/L, 0.5mmol/L, 1mmol/L, 5mmol/L, 10mmol/L, mol ratio
Can be 1~3：1~3, concretely 1~3：1、1：1 or 3：1.
Further, the concentration of the active metal salt can be 0.01~1mmol/L, concretely 0.01mmol/L,
0.05mmol/L, 0.1mmol/L, 0.5mmol/L, 1mmol/L, concretely 0.1mmol/L, 0.5mmol/L, 1mmol/L,
5mmol/L、10mmol/L。
Further, the concentration of the reducing agent can be 0.1~10mmol/L.
Further, in the step (2), the mixing time can be 1~48h, concretely 1h, 5h, 10h, 15h,
20h、25h、30h、35h、40h、48h；The reaction temperature can be able to be 0~100 DEG C for reaction temperature in 0~100 DEG C of water
In water, concretely 0~100 DEG C, 0 DEG C, 10 DEG C, 20 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C, 70 DEG C, 80 DEG C or 100 DEG C.
On the other hand, present disclosure also include by the above method be prepared based on self-assembled structures analogue enztme.
On the other hand, present disclosure also includes the self-assembled structures analogue enztme material that is based on available for following any
The application of kind：
1) biomolecule kit and test paper development；
2) biological tissue dyes；
3) pollutant process；
4) elctro-catalyst.
Beneficial effects of the present invention are：In preparation method provided by the invention, self assembly row can occur by what is prepared in advance
For the chemical substance aqueous solution in mix, two kinds of molecules are fitted together by Hyarogen-bonding, and ionizable metal salt is on its surface
Adsorb, in the presence of reducing agent, in-situ preparation metal nanoparticle, the resulting metal nano based on self-assembled structures
Grain has good peroxidase activity under the conditions of wide pH and temperature range, high ionic strength etc..The materials synthesis condition
It is gentle easy, low in raw material price, easily it is commercially exploited.The metal nanoparticle material based on self-assembled structures of the present invention is also
The numerous areas such as electro-catalysis, water process, biomedicine are can apply to, are had wide practical use.
In order to more fully understand and implement, the invention will now be described in detail with reference to the accompanying drawings.
Brief description of the drawings
Fig. 1 is the transmission electron microscope photo based on self-assembled structures analogue enztme prepared by embodiment 1.
Fig. 2 is the rate of catalysis reaction based on self-assembled structures analogue enztme under condition of different pH prepared by embodiment 1, its
Middle a, b, c, d, e are respectively rate of catalysis reaction when ph is 4,5,6,7,8.
Fig. 3 is the rate of catalysis reaction based on self-assembled structures analogue enztme at different temperatures prepared by embodiment 1, wherein
A, B, C, D, E, F, G are respectively rate of catalysis reaction when reaction temperature is 25,37,45,55,65,75,85 DEG C.
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material used, reagent etc., unless otherwise specified, are commercially obtained in following embodiments.
【Embodiment 1】Preparation and its Mimetic Peroxidase active testing based on self-assembled structures analogue enztme
100mL melamines (5mmol/L) aqueous solution and 100mL cyanuric acids (5mmol/L) aqueous solution are mixed, added
Enter 4mL gold chlorides (10g/L), add 1mL sodium citrates (50mmol/L), stirring at normal temperature 24 hours, obtain being based on self assembly
The aqueous solution of structure nano gold.
1) sign based on self-assembled structures nanogold：
Detected with Japanese transmission electron microscope (JEM-2100F) and obtain being based on self-assembled structures nanogold under above-mentioned condition
Pattern, as shown in Figure 1, the results showed that manufactured in the present embodiment to be existed based on self-assembled structures nanogold pattern for little particle nanogold
High degree of dispersion shape on carrier.
2) the Mimetic Peroxidase determination of activity based on self-assembled structures nanogold：
0.1g is taken based on self-assembled structures nanogold by prepared by embodiment 1, ultrasonic disperse takes in 100mL water
0.1mL above-mentioned solution is added to 0.9mL differences pH buffer system (including 0.05mol/L Tris-Ac, 1mmol/L
H2O2,1mmol/L 2,2- join nitrogen-two (3- ethyls-benzothiazole -6- sulfonic acid) di-ammonium salts (ABTS)) in, measure is based on from group
Catalytic rate of the assembling structure nanogold in 37 DEG C of different pH systems, as shown in Fig. 2 based on self-assembled structures nanogold in pH 4
~8 scopes have very high rate of catalysis reaction；
0.1g is taken based on self-assembled structures nanogold by prepared by embodiment 1, ultrasonic disperse takes in 100mL water
0.1mL above-mentioned solution is added to 0.9mL pH=7 buffer system (including 0.05mol/L Tris-Ac, 1mmol/L
H2O2,1mmol/L2,2- connection nitrogen-two (3- ethyls-benzothiazole -6- sulfonic acid) di-ammonium salts (ABTS)) in, measure is based on self assembly
The catalytic rate of structure nano gold at different temperatures, as shown in figure 3, based on self-assembled structures nanogold in 25~85 DEG C of scopes
With very high rate of catalysis reaction；
0.1g is taken based on self-assembled structures nanogold by prepared by embodiment 1, ultrasonic disperse takes in 100mL water
0.1mL above-mentioned solution is added to 0.9mL pH=7 buffer system (including 0.05mol/L Tris-Ac, 1mmol/L
H2O2,1mmol/L2,2- connection nitrogen-two (3- ethyls-benzothiazole -6- sulfonic acid) di-ammonium salts (ABTS)) in, measure is based on self assembly
Catalytic rate of the structure nano gold under the different NaCl concentrations at a temperature of 37 DEG C, based on self-assembled structures nanogold very high
Under salt ionic concentration (5mol/L), its rate of catalysis reaction is unaffected.
【Embodiment 2】Preparation and its Mimetic Peroxidase active testing based on self-assembled structures analogue enztme
100mL melamines (5mmol/L) aqueous solution and 100mL ellagic acids (5mmol/L) aqueous solution are mixed, added
4mL gold chlorides (10g/L), 1mL sodium citrates (50mmol/L) are added, stirring at normal temperature 24 hours, obtain being based on self assembly knot
The aqueous solution of structure nanogold.
Mimetic Peroxidase active testing based on self-assembled structures nanogold：Analogue enztme is in wide pH, temperature, salt ion
Concentration range is same as Example 1, and the result of gained is also similar to that embodiment 1.
【Embodiment 3】Preparation and its Mimetic Peroxidase active testing based on self-assembled structures analogue enztme
100mL cyanuric acids (5mmol/L) aqueous solution and 100mL citric acids (5mmol/L) aqueous solution are mixed, added
4mL gold chlorides (10g/L), 1mL sodium citrates (50mmol/L) are added, stirring at normal temperature 24 hours, obtain being based on self assembly knot
The aqueous solution of structure nanogold.
Mimetic Peroxidase active testing based on self-assembled structures nanogold：Analogue enztme is in wide pH, temperature, salt ion
Concentration range is same as Example 1, and the result of gained is also similar to that embodiment 1.
【Embodiment 4】Preparation and its Mimetic Peroxidase active testing based on self-assembled structures analogue enztme
100mL cyanuric acids (5mmol/L) aqueous solution and 100mL trimesic acids (5mmol/L) aqueous solution are mixed, added
Enter 4mL gold chlorides (10g/L), add 1mL sodium citrates (50mmol/L), stirring at normal temperature 24 hours, obtain being based on self assembly
The aqueous solution of structure nano gold.
Mimetic Peroxidase active testing based on self-assembled structures nanogold：Analogue enztme is in wide pH, temperature, salt ion
Concentration range is same as Example 1, and the result of gained is also similar to that embodiment 1.
The invention is not limited in above-mentioned embodiment, if the various changes or deformation to the present invention do not depart from the present invention
Spirit and scope, if these changes and deformation belong within the scope of the claim and equivalent technologies of the present invention, then this hair
It is bright to be also intended to comprising these changes and deformation.
Claims (9)
1. a kind of preparation method based on self-assembled structures analogue enztme, comprises the following steps：
(1) chemical dissolution of self assembly behavior will can occur in water；The chemical substance that self assembly behavior can occur
For at least one of following substances combination：Melamine and cyanuric acid, melamine and ellagic acid, melamine and adjacent benzene two
Phenol -3,5- disulfonic acid, melamine and citric acid, melamine and trimesic acid, melamine and trithiocyanuric acid, melamine
Acid and citric acid, cyanuric acid and trimesic acid；
(2) active metal salt solution, reducing agent are added in above-mentioned (1), stirring, the self-assembled structures that are based on is produced and simulates
Enzyme.
2. preparation method according to claim 1, it is characterised in that：The active metal salt is at least one in following substances
Kind：Copper sulphate, copper chloride, copper acetate, copper nitrate, silver nitrate, gold chloride, platinum chloride, palladium bichloride, palladium nitrate, iron chloride, nitric acid
Iron, frerrous chloride, ferric sulfate, cobalt chloride, cobalt nitrate, nickel nitrate, nickel chloride.
3. preparation method according to claim 2, it is characterised in that：The reducing agent is at least one of following substances：
Sodium borohydride, potassium borohydride, ascorbic acid, sodium citrate, citric acid, ellagic acid, catechol -3,5- disulfonic acid.
4. preparation method according to claim 3, it is characterised in that：The chemical substance that self assembly behavior can occur
Concentration is 0.01~10mmol/L, and mol ratio is 1~3：1~3.
5. preparation method according to claim 4, it is characterised in that：The concentration of the active metal salt be 0.01~
1mmol/L。
6. preparation method according to claim 5, it is characterised in that：The concentration of the reducing agent is 0.1~10mmol/L.
7. preparation method according to claim 6, it is characterised in that：In step (2), mixing time is 1~48h, stirring
Temperature is 0~100 DEG C.
8. according to method preparation any one of claim 1-7 based on self-assembled structures analogue enztme.
9. the application described in claim 8 based on self-assembled structures analogue enztme in following any aspect：
1) biomolecule kit and test paper development；
2) ELISA；
3) biological tissue dyes；
4) pollutant process；
5) elctro-catalyst.
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