Application of oxidized graphene modified by polyethylene glycol

A technology of polyethylene glycol and graphene, applied in the direction of biochemical equipment and methods, enzymes, enzymes, etc., can solve the problems of failing to meet the requirements of trypsin and trypsin stabilizers, and achieve enhanced activity and stability, Improve activity and synthesize simple effects

Inactive Publication Date: 2012-11-21
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are few active agents that can improve enzyme activity
Recently, the gold nanoparticles studied by Huang Qing, a professor of Applied Physics at the Chinese Academy of Sciences in Shanghai, can increase the activity of trypsin by about 10%, which is very good compared to other materials that greatly reduce the enzyme activity, but still fails to meet the requirements of pancreatic enzymes. The significant increase in protease activity failed to meet the needs of its biological application, but almost no stabilizers for trypsin were found (except for immobilized enzymes)

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  • Application of oxidized graphene modified by polyethylene glycol
  • Application of oxidized graphene modified by polyethylene glycol
  • Application of oxidized graphene modified by polyethylene glycol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation of trypsin specific activator GO-10k- br -PEG-NH 2

[0022] Use 1g of expanded graphite as the initial raw material, add about 30 times the weight of sodium chloride solid and grind until there are no obvious visible particles, add water and suction filter, wash away the sodium chloride, and dry the finely ground graphite for later use. Add about 30mL of concentrated sulfuric acid (98.3%) into the flask, put in dried fine graphite, stir at room temperature for 8 hours, then place the mixture in an ice bath, add about 3g of potassium permanganate, and slowly raise the temperature to 115 ℃, slowly add about 150mL of water, then add about 10mL of 30% hydrogen peroxide and stir, centrifuge to remove the upper aqueous solution, and wash repeatedly until the solution no longer precipitates. Sonicate 5 mL of graphene oxide suspension with a concentration of 3 mg / mL for 30 min to obtain a clear graphene oxide hydrosol, then add NaOH solid at a final con...

Embodiment 2

[0025] Embodiment 2: Study the influence of the polyethylene glycol-modified graphene oxide obtained in Embodiment 1 on trypsin activity.

[0026] Graphene oxide GO-10k- modified with polyethylene glycol obtained in Example 1 br -PEG-NH 2 Interact with trypsin, mix at room temperature for 16min, and then measure the activity of the enzyme, the specific method is: 20μl of 0.1mg / ml trypsin respectively with 20μl of sterile water, 0.15mg / mL, 0.2mg / mL of GO- 10k- br -PEG-NH 2Mix at 25°C for 16 minutes, add 10 μl of 5×PBS to supplement 50 μl, add 50 μl of 5 mg / ml substrate casein for enzymatic hydrolysis reaction at 40°C, 10 minutes, stop the reaction with 100 μl of TCA (0.4M), centrifuge, Take 100 μl supernatant and 100 μl folinol (1:1) to 500 μl Na 2 CO 3 (0.4M) 40°C, 20min color reaction, measure OD 680 .

[0027] The result is as image 3 Shown: The enhancement of trypsin activity was concentration-dependent, when GO-10k- br -PEG-NH 2 When the final concentrations wer...

Embodiment 3

[0031] Example 3: Study the influence of the polyethylene glycol-modified graphene oxide obtained in Example 1 on the thermal stability of trypsin.

[0032] Trypsin and GO-10k- br -PEG-NH 2 The mixed solution was denatured at 70°C and 80°C for 5 minutes respectively, and then the activity of trypsin was measured according to the above enzyme activity assay method, specifically:

[0033] (1) When casein was used as the substrate, trypsin and GO-10k- br -PEG-NH 2 Mix according to the mass volume ratio (mg / mL) of 1:2. In addition, trypsin and sterile water, trypsin and 10k- br -PEG-NH 2 [Mass volume ratio (mg / mL) is 1:10] also mixed with each other, and then the mixture was placed at 70°C and 80°C for 5 minutes, and then the activity of trypsin was measured according to the above method. The result is as Figure 5 Shown: No GO-10k- br -PEG-NH 2 In this case, trypsin is almost denatured and has no activity. However, in GO-10k- br -PEG-NH 2 Under the protective effect o...

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Abstract

The invention relates to an application of oxidized graphene modified by polyethylene glycol as a trypsin activator and a stabilizer; the oxidized graphene modified by polyethylene glycol comprises oxidized graphene; the surface of the oxidized graphene is grafted with polyethylene glycol; and the oxidized graphene and polyethylene glycol are connected through amide bonds; wherein the precursor of the polyethylene glycol is polyethylene glycol with six branched chains (with a chain end of an amino group) with a molecular weight of 9000-11000; and the weight percent of the polyethylene glycol is 35-45%; the oxidized graphene modified by polyethylene glycol has a dimension of 10 nm-30 nm, and a height of 1 nm-2 nm. The oxidized graphene modified by polyethylene glycol of the invention is simple in synthesis, and can specifically improve the activity and thermal stability of trypsin; the improvement of the activity and thermal stability of trypsin can meet biomedical requirements.

Description

technical field [0001] The invention relates to a polyethylene glycol-modified graphene oxide that can be used as a trypsin activator. Background technique [0002] Trypsin is a kind of serine protease, which is an important digestive enzyme in higher animals and human body. Trypsin is synthesized in the form of zymogen in the pancreas, secreted as a component of pancreatic juice, and then decomposed by enterokinase or trypsin to become biologically active trypsin. It is an endopeptidase that selectively hydrolyzes peptide chains formed by carboxyl groups of lysine or arginine in proteins (polypeptide chains). [0003] In the body, trypsin not only acts as a digestive enzyme, but also acts on the precursors of other enzymes (such as chymotrypsinogen, carboxypeptidase, phospholipase, etc.) and activates them. Clinically, trypsin is often used to treat local edema, hematoma and abscess caused by empyema, hemothorax, ulcer, wound inflammation, traumatic injury, fistula, etc. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/36
Inventor 刘庄彭睿金丽玲杨凯姚凯张帅
Owner SUZHOU UNIV
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