Method for preparing magnetic covalent immobilized enzyme carriers

An immobilized enzyme carrier, covalent technology, applied in the field of biochemistry, can solve problems such as affecting enzyme activity

Active Publication Date: 2013-04-24
杭州馨海生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immobilization condition is to react the carrier and the enzyme at 30°C for 6 hours, which will affect the activity of the enzyme to some extent.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation method of magnetic covalently immobilized enzyme carrier, the method is as follows:

[0027] Preparation of oleic acid-coated Fe 3 o 4 Nanoparticles: pass into nitrogen, add 94g FeCl after exhausting the air in the three-necked flask 3 ?6H 2 O, 50.5g FeSO 4 ?7 H 2 O, 100g potassium oleate (chemically pure) and 100ml water. It was placed in a constant temperature stirring tank at 80°C and stirred for 30 minutes. After the stirring was completed, NH 3 ?H 2 O solution (1M) until the solution pH = 11, then continue stirring for 30 min. After the reaction, the magnetic particles are separated using a magnetic field. Wash the magnetic particles with ethanol and deionized distilled water until the pH of the washing solution = 7, and then the black oleic acid-coated Fe 3 o 4 Nanoparticles, and then freeze-dry the obtained nanoparticles with a freeze dryer for future use.

[0028] Fe 3 o 4 Copolymerization reaction with enoic acid ester: Introduce ni...

Embodiment 2

[0031] The preparation method of magnetic covalently immobilized enzyme carrier, the method is as follows:

[0032] Preparation of oleic acid-coated Fe 3 o 4 Nanoparticles: pass into nitrogen, discharge the air in the three-necked flask, then add 23.5g FeCl 3 ?6H 2 O, 10.1 g FeSO 4 ?7 H 2 O, 30g potassium oleate and 50ml water. It was placed in a constant temperature stirring tank at 75 °C and stirred for 1 hour. Add a certain amount of 1 M sodium hydroxide solution until the pH of the solution = 11, and then continue to stir for 1 hour. After the reaction, the magnetic particles are separated using a magnetic field. Wash the magnetic particles with ethanol and deionized distilled water until the pH of the washing solution = 7, and then the black oleic acid-coated Fe 3 o 4 Nanoparticles. Then it was freeze-dried with a freeze dryer for future use.

[0033] Fe 3 o 4 Copolymerization reaction with enoic acid ester: Introduce nitrogen, discharge the air in the three-...

Embodiment 3

[0036] The preparation method of magnetic covalently immobilized enzyme carrier, the method is as follows:

[0037] Preparation of oleic acid-coated Fe 3 o 4 Nanoparticles: pass into nitrogen, discharge the air in the three-necked flask, then add 1.04g FeCl 3 ?6H 2 O, 0.48g FeSO 4 ?7 H 2 O, 1g potassium oleate (chemically pure) and 10ml water. It was placed in a constant temperature stirring tank at 80°C and stirred for 40 minutes. A certain amount of potassium hydroxide solution with a concentration of 2M was added until the pH of the solution = 13, and then continued to stir for 40 minutes. After the reaction, the magnetic particles are separated using a magnetic field. Wash the magnetic particles with ethanol and deionized distilled water until the pH of the washing solution = 7, and then the black oleic acid-coated Fe 3 o 4 Nanoparticles. It was directly used in the following reaction without lyophilization.

[0038] Fe 3 o 4 Copolymerization reaction with eno...

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PUM

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Abstract

The invention discloses a method for preparing magnetic covalent immobilized enzyme carriers, which relates to the field of biological chemistry and comprises the following steps that: firstly, oleic acid covered Fe3O4 with a magnetic structure is prepared, and then, Fe3O4 carries out polymerization reaction with olefine acid ester and subsequently and sequentially carries out reaction with hydrazine hydrate and nitrous acid to obtain activated immobilized enzyme carriers in nitrine magnetic nanometer structures. The method of the invention has the advantages that the process is simple, the operation cost is low, the enzyme carrying quantity is high, the stability is good, the reaction is easy to control and operate, the enzyme carrying capacity is as high as 250mg / g, and the original enzyme performance of the immobilized enzyme can almost be maintained at 100 percent.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a method for preparing a magnetic covalently immobilized enzyme carrier. Background technique [0002] Immobilized enzyme refers to an enzyme preparation that is not easy to lose with water, that is, its movement is restricted, and can play a catalytic role after being treated by physical or chemical methods. Compared with free enzymes, the advantages of immobilized enzymes are: it can be used multiple times, and the stability of the enzyme is improved; after the reaction, the immobilized enzyme is easy to separate from the substrate and product; the reaction conditions are easy to control, which is conducive to automated production. [0003] Magnetic nanostructured Fe 3 o 4 It has attracted extensive attention because of its biocompatibility, small size effect, large surface area, and special magnetic properties. Fe 3 o 4 The literature on covalent immobilization of enzymes as c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/08
Inventor 于洪巍王丹王珏王博刘敏
Owner 杭州馨海生物科技有限公司
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