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Preparation and application of dopamine functional magnetic nano-carrier

A magnetic nanocarrier, magnetic nanoparticle technology, applied to magnetic objects, immobilized on or in inorganic carriers, magnetic materials, etc., can solve the problems of complicated operation, influence of enzyme activity, adding coupling agent, etc., and achieve good immobilization. improved efficiency, reduced swing freedom restrictions, and improved dynamism

Inactive Publication Date: 2016-05-04
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these immobilization methods are not only cumbersome to operate, but also need to add a coupling agent, which has a certain impact on the activity of the enzyme and needs to be improved.

Method used

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  • Preparation and application of dopamine functional magnetic nano-carrier
  • Preparation and application of dopamine functional magnetic nano-carrier
  • Preparation and application of dopamine functional magnetic nano-carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: immobilized lipase

[0042] Weigh 6.0gFeSO 4 4H 2 O and 11.6 g FeCl 3 6H 2 O was dissolved in 350mL of distilled water, then transferred to a three-neck round bottom flask, and at 80°C, under the conditions of high-speed stirring and nitrogen protection, 20mL of 25% NH 4 Oh. After reacting for 5 minutes, 1.0 mL of oleic acid was added. The black precipitate obtained after stirring and reacting for 25 minutes was repeatedly washed with absolute ethanol and ultrapure water until the supernatant was colorless and transparent, precipitated and separated by a strong magnet, and dried in a vacuum oven at 50°C to obtain oleic acid magnetic nanoparticles called Fe 3 o 4 OA.

[0043] 0.2 g of dopamine hydrochloride was weighed and dissolved in 10 mM Tris-HCl solution at pH 8.5 to prepare a 2 mg / mL dopamine hydrochloride solution. Grind 1.0 g of the above-prepared oleic acid magnetic nanoparticles into 100 mL of dopamine hydrochloride solution evenly, ultr...

Embodiment 2

[0048] Embodiment 2: immobilized penicillin G acylase

[0049] The preparation of dopamine-functionalized magnetic nanocarrier is the same as that in Example 1.

[0050] Weigh 6 parts of dopamine-functionalized magnetic nanoparticles, 30 mg each, add penicillin G acylase solution with concentrations of 1.37, 2.74, 4.11, 5.48, 6.85, and 8.22 mg / mL, and fix at pH 6.0 at 25 °C After reaction for 8 hours, after the reaction, centrifuge, take 0.5mL of supernatant, use Tris-HCL buffer solution (0.1M, pH9.0) to wash twice, take 0.5mL of supernatant respectively, and use Bradford method to measure the first 3 The content of protein in the supernatant was measured by PDAB method for the enzyme activity of the above-mentioned immobilized penicillin G acylase.

[0051] Depend on Figure 4 , it can be seen that at different enzyme concentrations, Fe 3 o 4 The effect of OADP immobilization of lipase showed a significant difference. When the enzyme concentration is in the range of 1....

Embodiment 3

[0052] Example 3: Immobilized cellulase

[0053] The preparation of dopamine-functionalized magnetic nanocarrier is the same as that in Example 1.

[0054] Weigh 6 parts of dopamine-functionalized magnetic nanoparticles, each 30 mg, add cellulase solution with a concentration of 0.74, 1.48, 2.22, 2.96, 3.7, 4.44 mg / mL respectively, and immobilize the reaction at pH 5.0 and 25 °C After 10 hours of reaction, centrifuge, take 0.5mL of supernatant, wash twice with citrate buffer solution (0.1M, pH5.0), take 0.5mL of supernatant respectively, and use Bradford method to measure the three times before The content of protein in the supernatant, and the enzymatic activity of immobilized cellulase were determined by DNS method.

[0055] Depend on Figure 5 , it can be seen that at different enzyme concentrations, Fe 3 o 4 The effect of OADP immobilization of lipase showed a significant difference. When the enzyme concentration was in the range of 0.74-4.44mg / mL, the immobilizatio...

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Abstract

The invention belongs to the field of immobilization of proteins, and relates to preparation and application of a dopamine functional magnetic nano-carrier. The preparation comprises the following steps: firstly, dissolving FeSO4.4H2O and FeCl3.6H2O into distilled water, high-speed stirring and under protection of nitrogen, adding ammonia water of 25% NH4OH, and adding oleic acid; secondly, repeatedly cleaning black precipitates obtained by a stirring reaction and then using a strong magnet to perform precipitation separation, and drying to obtain oleic acid magnetic nano-particles; and finally, adding the oleic acid magnetic nano-particles into a dopamine hydrochloric acid solution, after the reaction is over, using a magnet to separate, cleaning and drying obtained deep brown precipitates, and then the dopamine functional magnetic nano-carrier can be obtained. According to the preparation and application of the dopamine functional magnetic nano-carrier provided by the invention, dopamine hydrochloride is adopted for modifying magnetic oleic acid nano-particles, so that the surface of a magnetic nano-particle has a plentiful of quinonyls and amidogen functional groups, thus proteins can be effectively immobilized; besides, the surface of the carrier has a layer of soft poly dopamine oxide films, the surface area is larger, and the preparation method is simple.

Description

technical field [0001] The invention belongs to the field of protein immobilization, and relates to the preparation and application of a dopamine functionalized magnetic nanometer carrier. Background technique [0002] Enzymes are mainly composed of proteins and have catalytic activity. Due to their high catalytic activity and strong substrate transfer, they have broad application prospects in the fields of food, brewing, medicine, environment and chemicals. However, the free enzyme has poor stability and is easily denatured and inactivated under extreme conditions such as high temperature, strong acid and strong alkali, and it is difficult to separate from the substrate and product after the catalytic reaction, resulting in product pollution and increased production costs. limit its industrial application. In order to solve the above shortcomings of free enzymes, the concept of immobilized enzymes was proposed in the 1960s. Enzyme immobilization refers to the process of ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): H01F1/00B82Y25/00C12N11/14
CPCH01F1/0045B82Y25/00C12N9/20C12N9/2437C12N9/84C12N11/14C12Y301/01003C12Y305/01011
Inventor 张业旺王进王香玉庄孟瑶凌小敏
Owner JIANGSU UNIV
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