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Graphene oxide modified by polyethylene glycol, and application of graphene oxide

A polyethylene glycol and graphene technology, applied in biochemical equipment and methods, enzymes, enzyme stabilization, etc., can solve problems such as failure to meet trypsin, trypsin stabilizer not found, etc., and achieve enhanced activity and stability. , improve the activity, the effect of simple synthesis

Inactive Publication Date: 2012-05-02
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)

Method used

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  • Graphene oxide modified by polyethylene glycol, and application of graphene oxide
  • Graphene oxide modified by polyethylene glycol, and application of graphene oxide
  • Graphene oxide modified by polyethylene glycol, and application of graphene oxide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Preparation of trypsin specific activator GO-2k- l -NH 2 -PEG

[0021] 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 conc...

Embodiment 2

[0024] Embodiment 2: Study the influence of the polyethylene glycol-modified graphene oxide obtained in Embodiment 1 on trypsin activity.

[0025] The polyethylene glycol-modified graphene oxide GO-2k- obtained in Example 1 l -PEG-NH 2Interact with trypsin, mix at room temperature for 16 minutes, and then measure the activity of the enzyme, see literature. (See: Lv Min, Su Y Y, et al. Preparative Biochemistry & Biotechnology, 2009, 39: 429–438). The specific method is: 20μl of 0.1mg / ml trypsin respectively mixed with 20μl of sterile water, 0.2mg / mL, 0.3mg / mL, 0.4mg / mL, 0.5mg / mL, 0.6mg / mL of GO-2k- l -NH 2 - PEG was mixed at 25°C for 16 minutes, then added 10 μl 5×PBS to make up to 50 μl, and then 50 μl of 5 mg / ml substrate casein was added for enzymatic hydrolysis reaction at 40°C for 10 minutes, and the reaction was terminated with 100 μl of TCA (0.4M). Centrifuge, take 100 μl supernatant and 100 μl folinol (1:1) into 500 μl Na 2 CO 3 (0.4M) 40°C, 20min color reaction, ...

Embodiment 3

[0030] Example 3: Study the influence of the polyethylene glycol-modified graphene oxide obtained in Example 1 on the thermal stability of trypsin.

[0031] Trypsin and GO-2k- l -NH 2 -The mixed solution of PEG 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:

[0032] (1) When casein was used as the substrate, trypsin and GO-2k- l -PEG-NH 2 Mix according to the mass volume ratio (mg / mL) of 1:2. In addition, trypsin and sterile water, trypsin and NH 2 -2k- l -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-mentioned method. The result is as Figure 5 Shown: No GO-2k- l -PEG-NH 2 In this case, trypsin is almost denatured and has no activity. However, in GO-2k- l -PEG-NH 2 Under the protecti...

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Abstract

The invention relates to graphene oxide modified by polyethylene glycol. The graphene oxide can be used as trypsin activator. The graphene oxide modified by the polyethylene glycol comprises graphene oxide and polyethylene glycol; one end of the polyethylene glycol is an amino group, and other end of the polyethylene glycol is an imino group, and nitrogen atoms of the imino group and carbon atomsin carbonyl of the graphene oxide are connected with each other to form an amido bond; the height of the graphene oxide is 1 to 2nm, and the size of the graphene oxide is 10 to 30nm; the polyethyleneglycol is a liner-chain polyethylene glycol of which the molecular weight is 1,800 to 2,200; and the graphene oxide comprises 25 to 35 weight percent of the polyethylene glycol. The graphene oxide modified by the polyethylene glycol is easy to synthesize, and can specifically improve the activity and the thermal stability of trypsin; and the activity and stability of the trypsin are improved to meet biomedical demand of the trypsin.

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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IPC IPC(8): C08G65/48C12N9/76C12N9/96
Inventor 彭睿刘庄金丽玲杨凯姚凯张帅李述汤
Owner SUZHOU UNIV
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