CN109603858B

CN109603858B - Double-activity mimic enzyme material and preparation and application thereof

Info

Publication number: CN109603858B
Application number: CN201811533227.7A
Authority: CN
Inventors: 王瑾; 王毅; 张盾
Original assignee: Institute of Oceanology of CAS
Current assignee: Institute of Oceanology of CAS
Priority date: 2018-12-14
Filing date: 2018-12-14
Publication date: 2021-11-16
Anticipated expiration: 2038-12-14
Also published as: CN109603858A

Abstract

The invention provides a preparation method and application of a cobalt disulfide nano material with double activities of a peroxide mimic enzyme and an oxide mimic enzyme. The method belongs to the technical field of mimic enzyme catalytic oxidation. The cobalt disulfide mimic enzyme material with high activation performance is obtained by a one-step hydrothermal method. The nano material prepared by the invention has the advantages of simple synthesis method, low cost, remarkable performance and the like, and has wide application prospect in the field of nano material catalytic oxidation.

Description

Double-activity mimic enzyme material and preparation and application thereof

Technical Field

The invention relates to a mimic enzyme technology, in particular to a double-activity mimic enzyme material and preparation and application thereof.

Background

The active enzyme in the organism has high specificity and catalytic activity, and is widely applied to a plurality of fields of biosensing, medical care, environmental protection, food industry and the like. However, most natural enzymes have poor stability and are easily inactivated by unavoidable conditions such as temperature, pH value and protease existence, and the preparation and purification costs are high, so that the application of the natural enzymes is greatly limited. Therefore, the research for developing the mimic enzyme material with simple preparation method, strong stability and low cost is very important.

In recent years, with the discovery that more and more nanomaterials or nanocomposites have been demonstrated to mimic enzymatic properties, such as MnOOH [1 ]]、CuS[2]、Fe₃S₄[3]And the like. Compared with the natural enzyme obtaining method, on one hand, the material cost is reduced, and on the other hand, the synthetic method is simple and convenient, and the unit operation is easy to realize.

1.Wang Y,Zhang D,Xiang ZB(2016)Synthesis and intrinsic enzyme-like activity ofβ-MnOOH nanoplates.J Taiwan Inst Chem Eng 59:547-552.

2.Yang ZJ,Cao Y,Li J,MM L,Jiang ZK,XY H(2016)Smart CuS nanoparticles as peroxidase mimetics for the Desigh of Novel Label-Free Chemiluminescent Immunoassy.ACS Appl Mater Interfaces 8:12031-12038.

3.Ding CP,Yan YH,Xiang DS,Zhang CL,Xian YZ(2016)Magnetic Fe₃S₄nanoparticles with peroxidase-like activity,and their use in a photometric enzymatic glucose assay.Microchim Acta 183:625-631.

Disclosure of Invention

The invention aims to provide a double-activity mimic enzyme material, and preparation and application thereof.

In order to achieve the purpose, the invention adopts the technical scheme that:

a method for preparing a double-activity mimic enzyme nano material comprises the step of obtaining a chemical component CoS by a one-step hydrothermal method₂Of (4) a nanomaterial of (2).

Further, cobalt salt and Na₂S₂O₃·5H₂Mixing O powder, dissolving in deionized water, uniformly mixing by magnetic stirring, and reacting for 24-28 hours under the conditions of 160 ℃ and 200 ℃ in a sealed manner after uniform mixing to obtain the cobalt disulfide mimic enzyme material; wherein, Co in the cobalt salt²⁺With Na₂S₂O₃·5H₂O is added according to the molar ratio of 2-1: 1-3.

And uniformly mixing the mixture by magnetic stirring for 20-40min, placing the mixture into a hydrothermal reaction kettle, covering and sealing the hydrothermal reaction kettle, placing the hydrothermal reaction kettle into a constant-temperature air-blast drying box, reacting for 24-28 hours at the temperature of 160-200 ℃, naturally cooling the reaction kettle to room temperature after the reaction, and sequentially cleaning and drying the product.

The cobalt salt is cobalt chloride hexahydrate or cobalt nitrate hexahydrate;

the Co²⁺The optimal molar ratio of the urea to the urea is 1: 1; co²⁺The molar ratio to thioacetamide is 1: 1.

and washing the product with deionized water and ethanol in sequence, performing centrifugal separation, and drying for 10-12h to obtain the target material.

The double-activity mimic enzyme material obtained by the preparation method is used for obtaining the cobalt disulfide nano material by a one-step hydrothermal method according to the method.

The application of the double-activity mimic enzyme material and the application of the cobalt disulfide nano material in oxidation-reduction reaction.

The cobalt disulfide nano material is used as a mimic enzyme to perform catalytic oxidation-reduction reaction on a substrate.

The cobalt disulfide nano material is used as mimic enzyme to carry out catalytic oxidation-reduction reaction on a substrate under an acidic condition.

The cobalt disulfide nano material is used as an oxide mimic enzyme or a peroxide mimic enzyme to perform catalytic oxidation-reduction reaction on a substrate under an acidic condition.

The substrates are TMB and H₂O₂Or TMB.

Compared with the prior art, the invention has the following advantages and prominent effects:

the invention obtains the cobalt disulfide-peroxide mimic enzyme and oxide mimic enzyme double-activity nano material by a one-step hydrothermal method. The material obtained by the invention has low cost, simple preparation method and good repeatability; the mimic enzyme can be used as a novel mimic enzyme and has potential application value in the fields of immunoassay, biological detection, clinical diagnosis and the like. It has wide application prospect in novel catalytic oxidation analysis.

Description of the drawings:

FIG. 1 is a TEM image of a nanomaterial provided by an embodiment of the present invention;

FIG. 2 is an X-ray diffraction pattern of a qualitatively analyzed material composition provided by an embodiment of the present invention;

fig. 3 is a colorimetric diagram of the nanomaterial provided in the embodiment of the present invention for simulating oxidase catalysis and peroxidase catalysis.

Detailed description of the inventionin the following, the invention is further illustrated by means of specific examples, which will assist the person skilled in the art in a more complete understanding of the invention, but without in any way limiting it.

Example 1:

adding the molar ratio into the beaker respectivelyCoCl of 1:1₂·6H₂O and Na₂S₂O₃·5H₂And O, dissolving in deionized water, and uniformly mixing by magnetic stirring. And transferring the reaction solution into a 100mL hydrothermal reaction kettle, covering and sealing the kettle, placing the kettle in a constant-temperature air-blowing drying oven, and reacting for 24 hours at 160 ℃. After the reaction is finished, the reaction kettle is naturally cooled to room temperature. The product is firstly centrifugally washed by ultrapure water for 3 times, and then centrifugally washed by absolute ethyl alcohol for 2-4 times; and centrifuging for 10min under the centrifugation condition of 4000 revolutions per minute, and drying for 10h at 60 ℃ after washing to obtain the cobalt disulfide mimic enzyme material. (see FIGS. 1 and 2).

The synthesized cobalt disulfide nano material can be seen from figure 1 as a lamellar structure with a certain geometric shape. The lamellar structure greatly increases the surface area of catalytic reaction and provides more active sites for catalytic oxidation. From FIG. 2, it can be seen that the synthesized cobalt disulfide nano material and components and CoS₂Standard cards (JCPDS No.41-1471) are matched.

Examples 2 to 5:

the preparation process comprises the following steps:

taking 4 beakers, respectively adding a certain amount of CoCl₂·6H₂O and Na₂S₂O₃·5H₂And O, dissolving in deionized water, and uniformly mixing by magnetic stirring. And transferring the reaction solution into a 100mL hydrothermal reaction kettle, covering and sealing the kettle, placing the kettle in a constant-temperature air-blowing drying oven, and reacting for 24 hours at 160 ℃. After the reaction is finished, the reaction kettle is naturally cooled to room temperature. The product is sequentially cleaned and dried to obtain the cobalt disulfide mimic enzyme material, and XRD test results show that the product is CoS₂(see Table 1)

TABLE 1

Examples	n(CoCl₂·6H₂O):n(Na₂S₂O₃·5H₂O)
		2	2:1
3	1:1
		4	1:2
5	1:3

Application example

At 0.2mM H₂O₂0.15mM TMB, 0.3mg/mL nanomaterial, pH 3.05 in PBS buffer. When only hydrogen peroxide and mimic enzyme materials are contained in the system and TMB is not added, the solution is colorless; when only TMB and mimic enzyme materials are in the solution and no hydrogen peroxide is added, the solution is slightly blue, which indicates that the nano material has oxidase property; when the mimic enzyme material was added to the TMB and hydrogen peroxide system, the solution turned a darker blue color; the nano material provided by the invention has double activities of peroxidase and oxidase.

Claims

1. A preparation method of a double-activity mimic enzyme nano material is characterized by comprising the following steps: obtaining CoS with chemical composition by one-step hydrothermal method₂The nano material of the lamellar structure of (a);

mixing cobalt salt and Na₂S₂O₃·5H₂Mixing O powder, dissolving in deionized water, magnetically stirring and uniformly mixing, and sealing and reacting at the temperature of 160 ℃ and 200 ℃ for 24-28h to obtain the cobalt disulfide mimic enzyme material; wherein, Co in the cobalt salt²⁺With Na₂S₂O₃·5H₂O is added according to the mol ratio of 2-1: 1-3;

2. The method of preparing the double-activity mimic enzyme nanomaterial of claim 1, wherein the method comprises the following steps: and washing the product with deionized water and ethanol in sequence, performing centrifugal separation, and drying for 10-12h to obtain the target material.

3. A dual-activity mimetic enzyme material produced by the process of claim 1, wherein: the method of claim 1, wherein the cobalt disulfide nanomaterial with a lamellar structure is obtained by a one-step hydrothermal method.

4. Use of a dual activity mimetic enzyme material according to claim 3, wherein: the lamellar structure cobalt disulfide nano material is applied to oxidation-reduction reaction.

5. Use of the double-active mimetic enzyme material according to claim 4, wherein: the lamellar structure cobalt disulfide nano material is used as a mimic enzyme to perform catalytic oxidation-reduction reaction on a substrate.

6. Use of a dual activity mimetic enzyme material according to claim 5, wherein: the lamellar structure cobalt disulfide nano material is used as a mimic enzyme to carry out catalytic oxidation-reduction reaction on a substrate under an acidic condition.

7. Use of a dual-activity mimetic enzyme material according to claim 6, wherein: the lamellar structure cobalt disulfide nano material is used as an oxide mimic enzyme or a peroxide mimic enzyme to perform catalytic oxidation-reduction reaction on a substrate under an acidic condition.

8. Use of a dual activity mimetic enzyme material according to claim 7, wherein: the substrates are TMB and H₂O₂Or TMB.