Method for immobilizing enzyme by using polydopamine-coated plastic filler

A technology of polydopamine and immobilized enzymes, applied in the direction of immobilized enzymes, biochemical equipment and methods, enzymes, etc., can solve the problems of substrates with specific requirements, short-term effects, difficult coating adhesion, etc., and increase the microscopic surface area , increase the fixed point, the effect of enhanced adhesion

Pending Publication Date: 2021-11-02
HEBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the surface energy of the plastic filler is low, and it is difficult to be adhered by the coating, which limits its application in the fields of coating and immobilized enzymes. Therefore, it is necessary to modify the surface of the plastic filler to have various required properties.
[0003] At present, most surface modification methods (such as surface roughening, chemical treatment, radiation, spraying treatment and plastic modification, etc.) have their own limitations, such as complex process, short-term effect, and harmful to the substrate. Specific requirements and unfriendly to the environment, etc.

Method used

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  • Method for immobilizing enzyme by using polydopamine-coated plastic filler
  • Method for immobilizing enzyme by using polydopamine-coated plastic filler
  • Method for immobilizing enzyme by using polydopamine-coated plastic filler

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Take 12.114 g of tris (Tris) in a 1 L volumetric flask, add deionized water to obtain a 0.1 M Tris buffer solution, and add hydrochloric acid to adjust the pH to 8.5. Dissolve 2 g of dopamine in Tris buffer solution to obtain a fresh dopamine solution with a concentration of 2.0 mg / mL. Put the polypropylene filler into the isopropanol solution, ultrasonically wash it for 30 minutes, and then ultrasonically clean it with distilled water for 15 minutes to remove the grease and dust on the surface, take it out and put it in a vacuum drying oven at 50°C. Immerse the cleaned and dried polypropylene packing in the dopamine solution, react continuously in a water bath constant temperature shaker at room temperature for 1.5h, take out the packing, wash thoroughly with ethanol and deionized water in turn until the filtrate is colorless and transparent, put it in 60℃ Dry overnight in a vacuum oven to obtain a dopamine-modified polypropylene filler.

[0039] Weigh 0.548 g of meth...

Embodiment 2

[0047] Other steps are the same as in Example 1, except that the mass ratio of MTMS to TMOS is replaced by 3:1, and the specific addition amounts are 0.651 g of methyl orthosilicate and 1.953 g of methyl trimethoxysilane.

[0048] The experimental results showed that the protein immobilization rate was 89.62±2.4%; the adhesive strength was 55.734MPa; after incubation in pH 10.0 for 12h, the relative enzyme activity was 80%; after incubation in the organic solvent ethyl acetate for 20h, the relative enzyme activity was 89%; 88% relative activity after storage for 60 days

[0049] It can be seen from the experimental results that the protein immobilization rate and adhesive strength do not change much after the ratio of silane precursor is changed, because the protein immobilization rate and adhesive strength mainly depend on the dopamine coating on the surface of the filler. The relative enzyme activity is slightly reduced, because the lipase CaLB is an interface active enzyme,...

Embodiment 3

[0051] Other steps are the same as in Example 1, except that the amount of methanol added is replaced by 60% of the total mass of liquid A, and the specific amount added is 3.65 g.

[0052] The experimental results showed that the protein immobilization rate was 88.42±2.6%; the adhesive strength was 56.762MPa; after incubation in pH 10.0 for 12 hours, the relative enzyme activity was 78%; after incubation in the organic solvent ethyl acetate for 20 hours, the relative enzyme activity was 89%; the relative enzyme activity was 79% after storage for 60 days.

[0053] From the experimental results, it can be seen that the enzyme activity decreased significantly. This is because methanol acts as a dispersant in the formula, allowing the enzyme to disperse evenly, but methanol damages the activity of the enzyme and reduces the activity of the enzyme, so the relative activity decreases significantly.

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Abstract

The invention relates to a method for immobilizing enzyme by using polydopamine-coated plastic filler. The method comprises the following steps: putting a plastic filler into a dopamine solution with a proper concentration, carrying out a constant-temperature oscillation reaction, carrying out surface modification to enable the plastic filler to have better adhesion, wettability and surface roughness, and loading sol containing an immobilized enzyme solution on the modified filler. The method is low in cost, good in effect and easy and convenient to operate, and the obtained plastic enzyme-loaded filler has higher acid-base stability, organic solvent stability, storage stability and reusability.

Description

Technical field: [0001] The method for immobilizing enzymes involved in the invention is a method for modifying the surface of plastic fillers and applying it to the fields of coating and immobilizing enzymes. Background technique: [0002] Plastic packing has been widely used in various packing towers due to its advantages of light weight, cheap price, high temperature resistance, corrosion resistance and low damage rate. However, the surface energy of the plastic filler is low, and it is difficult to be attached by the coating, which limits its application in the fields of coating and immobilized enzymes. Therefore, it is necessary to modify the surface of the plastic filler to make it have various required properties. [0003] At present, most surface modification methods (such as surface roughening, chemical treatment, radiation, spraying treatment and plastic modification, etc.) have their own limitations, such as complex process, short-term effect, and harmful to the s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/082C12N11/089C08J7/04C08L23/12C08L23/06C08L27/06
CPCC12N11/082C12N11/089C12N9/20C08J7/042C12Y301/01003C08J2323/12C08J2327/06C08J2323/06C08J2479/04
Inventor 王洪海张宇尹伊岳文达
Owner HEBEI UNIV OF TECH
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