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Preparation method of polydopamine/graphene oxide composite film with muramidase immobilized

A technology of graphene composite and polydopamine film, which is applied in the direction of fixation on or in the inorganic carrier, can solve the problems of short protein activity, affecting use, poor stability, etc., and achieve antibacterial time improvement, good immobilization, and antibacterial performance Improved effect

Inactive Publication Date: 2016-07-06
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still some problems, such as the short activity and poor stability of the protein, which greatly affects its use.

Method used

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  • Preparation method of polydopamine/graphene oxide composite film with muramidase immobilized
  • Preparation method of polydopamine/graphene oxide composite film with muramidase immobilized
  • Preparation method of polydopamine/graphene oxide composite film with muramidase immobilized

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Example 1: The PDA / rGO-cLY film was modified with glass as the substrate.

[0045]The reaction raw materials are dopamine hydrochloride, Tirs solution at pH 8.5, graphene oxide, egg white lysozyme, and PBS buffer at pH 7.4. Among them, dopamine hydrochloride was purchased from Sigma-Aldrich, egg white lysozyme was purchased from Sigma-Aldrich, the activity unit was ~100000U / mg, the molecular weight was a single chain of 14.3kDa, and the graphene oxide was obtained by the Hummers method (Marcano, D.C.; Kosynkin, D.V.; Berlin, J.M.; Sinitskii, A.; Sun, Z.; Slesarve, A.; Alemany, L.B.; Lu, W.; Tour, J.M. Improved Synthesis of Graphene Oxide.

[0046] (1) Dopamine modification of the glass surface

[0047] Dopamine was added to Tris solution at pH 8.5 to prepare 2 mg / mL, put into the cleaned glass slide, let stand at room temperature for 24 hours, take it out, ultrasonically clean it, and dry it with nitrogen gas.

[0048] (2) Preparation of incomplete graphene derivati...

Embodiment 2

[0070] Example 2: The glass surface was modified with PDA / rGO-cLY film.

[0071] The reaction raw materials are dopamine hydrochloride, Tirs solution with pH 8.5, egg white lysozyme, and PBS buffer solution with pH 7.4. Among them, dopamine hydrochloride was purchased from Sigma-Aldrich, egg white lysozyme was purchased from Sigma-Aldrich, the activity unit was ~100000U / mg, and the molecular weight was a single chain of 14.3kDa. Graphene oxide was prepared by the Hummers method. Fluorescein isothiocyanate (FITC) was purchased from Sigma-Aldrich.

[0072] Lysozyme was labeled with fluorescein isothiocyanate (FITC). Prepare 100 mL of a round solution of carbonate and sodium chloride with a pH of 9, add 1 g of lysozyme, disperse and dissolve, and lower the temperature to 4°C. Add 10 mg FITC to the lysozyme solution at 4°C, and magnetically stir in an ice-water bath for 12 hours in the dark to generate fluorescently labeled protein FITC-cLY. Prepare excess half-saturated amm...

Embodiment 3-5

[0080] Embodiment 3-5: The PDA / rGO film layer was prepared on the glass surface under different concentration conditions.

[0081] The reaction raw materials are dopamine hydrochloride, Tirs solution with pH 8.5, and graphene oxide. Among them, dopamine hydrochloride was purchased from Sigma-Aldrich, and graphene oxide was obtained by Hummers method.

[0082] (1) Dopamine modification of the glass surface

[0083] Dopamine was added to Tris solution at pH 8.5 to prepare 2 mg / mL, put into the cleaned glass slide, let stand at room temperature for 24 hours, take it out, ultrasonically clean it, and dry it with nitrogen gas.

[0084] (2) Preparation of incomplete graphene derivative film layer

[0085] Add the Tris solution at pH 8.5 to graphene oxide, prepare 0.1mg / mL, 0.3mg / mL and 0.5mg / mL graphene oxide solutions respectively, adjust the pH to 8.5, and put the dopamine-treated glass slide into In the graphene oxide solution, soak for 3 hours at 50°C, take it out, clean it...

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Abstract

The invention discloses a preparation method of a polydopamine / graphene oxide composite film with muramidase immobilized.The method includes the steps that firstly, autopolymerization of dopamine is carried out on the surface of a substrate like glass, an incomplete graphene oxide film layer is introduced onto a polydopamine film layer, part of the zone of the polydopamine film layer is covered with graphene oxide by controlling the concentration of graphene oxide and the temperature of the soaking process, the oxygen-containing functional group part of graphene oxide is reduced by polydopamine at the same time in the process, and a partially-reduced graphene oxide film layer is formed; finally, muramidase is immobilized to the incomplete partially-reduced graphene oxide film layer, and the polydopamine / graphene oxide composite film with muramidase immobilized is obtained.The method is simple and easy to implement; by inserting the incomplete graphene oxide film layer, the enzyme loading capacity of the composite film is obviously increased, the antimicrobial property is remarkably improved, and long-term high-efficiency bacteriostasis can be achieved; the composite film has good application prospects in the fields of food safety, medical treatment and public health and the like.

Description

technical field [0001] The invention relates to the field of preparation methods of graphene oxide composite materials, in particular to a preparation method of a polydopamine / graphene oxide composite film with high antibacterial performance and enzyme loading capacity. Background technique [0002] Antibacterial glass or packaging bags, as a new type of ecological functional material, can be widely used in the fields of medical treatment, food safety and public health. The common antibacterial glass now uses ionized metals such as zinc, silver, copper, etc. to be added during the glass manufacturing process, or titanium dioxide film is coated on the glass surface, and the antibacterial effect is achieved through the photocatalysis of titanium dioxide. But these fungicides still have certain harm to the human body, and release into the environment will cause secondary pollution. Therefore, it is urgent to choose an antifouling agent that is more environmentally friendly and...

Claims

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Application Information

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IPC IPC(8): C12N11/14C03C17/42
CPCC12N11/14C03C17/42
Inventor 陈守刚郝湘平朱弘政王龙强岳龙飞
Owner OCEAN UNIV OF CHINA
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