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Preparation method and application of functionalized graphene oxide carrier

A graphene and functionalization technology, applied in chemical instruments and methods, immobilized on or in inorganic carriers, carrier-bound/immobilized peptides, etc., can solve the structural modification of graphene oxide, hinder the widespread use of enzymes, and cannot be reused and other problems, to achieve the effect of increasing the degree of freedom of swing, wide application range and strong applicability

Inactive Publication Date: 2015-05-27
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, free enzymes are difficult to separate from reaction products and cannot be reused, resulting in high production costs and low economic benefits, which seriously hinder the widespread use of enzymes.
There is no research report on the structural modification of graphene oxide by this method

Method used

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  • Preparation method and application of functionalized graphene oxide carrier
  • Preparation method and application of functionalized graphene oxide carrier
  • Preparation method and application of functionalized graphene oxide carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Immobilized cellulase

[0038] Weigh 1g of graphite powder and dissolve it in 25ml of concentrated sulfuric acid (ice bath), add 1g of NaNO 3 , 3 g KMnO 4 , after which the mixture was heated to 35 o C, continue stirring for 30 min, add 50 ml of distilled water, raise the temperature of the reactant to 90°C, continue stirring for 1 h, add 40 ml of distilled water and 3 ml of 30% hydrogen peroxide to terminate the reaction, and The reactant was filtered and washed with distilled water, and the product was resuspended in distilled water for 5 min and then vacuum-dried at 40°C to obtain graphene oxide.

[0039] figure 1 SEM and TEM images of graphene oxide prepared above. From the SEM and TEM images, it can be seen that graphene oxide has a sheet structure with a large surface area, wrinkles of different degrees, and a large protein load.

[0040] Weigh 0.5 g SESA and dissolve in 5 ml distilled water at 40 °C with 1.0 M Na 2 CO 3 Adjust the pH of the so...

Embodiment 2

[0044] Example 2 Immobilized penicillin G acylase

[0045] The preparation of the functionalized graphene oxide carrier is the same as in Example 1.

[0046] Weigh 11 parts of functionalized graphene oxide, each 25mg, add 0.3ml of liquid penicillin G acylase, then add 0.7ml of phosphate buffer (0.1 M, pH 6.0), react for 1min, 2min, 3min, 4min respectively , 5min, 7min, 10min, 15min, 20min, 30min, 60min, after the reaction, centrifuge, take 0.8ml of supernatant, then add 1ml of phosphate buffer (0.1 M, pH 6.0), centrifuge, take 0.9ml of supernatant ml, then add 1ml of citric acid buffer (0.1 M, pH 6.0), centrifuge, remove all the supernatant, use the Bradford method to measure the protein content in the first two supernatants respectively, and use the PDAB method to determine the above-mentioned immobilized penicillin G Acylase activity.

[0047] Depend on Figure 4 , it can be seen that with the increase of immobilization time, the immobilization yield and immobilization ...

Embodiment 3

[0048] Example 3 Immobilized bovine serum albumin

[0049] The preparation of the functionalized graphene oxide carrier is the same as in Example 1.

[0050] Weigh 11 parts of functionalized graphene oxide, 25 mg each, add 0.8 ml of bovine serum albumin, then add 0.2 ml of phosphate buffer (0.1 M, pH 7.0), and react for 1 min, 2 min, 3 min, 4 min, 5 min, respectively. 7min, 10min, 15min, 20min, 30min, 60min, after the reaction, centrifuge, take 0.8ml of supernatant, then add 1ml of phosphate buffer (0.1 M, pH 7.0), centrifuge, take 0.8ml of supernatant, and then Add 1ml of citric acid buffer (0.1 M, pH 7.0), centrifuge to remove all the supernatant, and use the Bradford method to measure the protein content in the first two supernatants respectively.

[0051] Depend on Figure 5 It can be seen that within 10 min, with the increase of immobilization time, the immobilization yield of bovine serum albumin increases. When the immobilization time reaches 10 min, as the time con...

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Abstract

The invention belongs to the field of protein immobilization and discloses preparation of a functionalized graphene oxide carrier and an application method of the functionalized graphene oxide carrier in rapid immobilization of proteins. According to the technical scheme, the invention has the key points of preparing and modifying functionalized graphene oxide; P-beta-sulfatoethylsulfonyl aniline (SESA) is adopted to activate the graphene oxide (GO), diazotization is carried out on the activated carrier, and finally, the carrier is used for rapidly immobilizing the proteins. The functionalized graphene oxide carrier has a lamellar structure and is large in surface area and simple in preparation method. According to the preparation method and the application provided by the invention, three different proteins in different shapes are used for immobilization, the immobilization can be rapidly finished with the functionalized graphene oxide as an immobilization carrier, and in addition, the immobilization yield and the immobilization efficiency can both reach 80-90 percent. Therefore, the method of preparing the functionalized graphene oxide carrier for immobilizing the proteins provides a new thinking to study rapid immobilization and has certain study significance.

Description

technical field [0001] The invention belongs to the field of protein immobilization, and relates to the preparation of a functionalized graphene oxide carrier and its application in rapid protein immobilization. Background technique [0002] Since the advent of immobilization technology, immobilization technology is developing at an unprecedented speed and is constantly improving. Among them, immobilized enzyme is an enzyme application technology developed in the past ten years, which has attractive application prospects in industrial production, chemical analysis and medicine (see: Guo Liquan, Wang Hongyu, Research Progress in Enzyme Immobilization Technology, Jilin Business School Journal, 2008). Enzyme, as a kind of macromolecular substance with biocatalytic function, is mostly protein. Enzymes are used as catalysts to catalyze reactions with high specificity, high efficiency and mild reaction conditions. However, free enzymes are difficult to separate from reaction pr...

Claims

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Application Information

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IPC IPC(8): C07K17/14C12N11/14C01B31/04
Inventor 张业旺李越江晓萍刘彩虹
Owner JIANGSU UNIV
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