Preparation method for separating lysozyme by amino-functionalized magnetic nanoparticles

A technology of magnetic nanoparticles and amino functionalization, applied in biochemical equipment and methods, chemical instruments and methods, enzymes, etc., can solve problems such as backward production technology and lysozyme scale gap, and achieve small impact, large adsorption capacity, Simple effect of extraction separation method

Inactive Publication Date: 2016-01-27
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, compared with foreign countries, there is still a large gap in the scale of my country's lysozyme manufacturing industry, and the production technology is still relatively backward
Therefore, the current domestic production and supply of highly active egg white lysozyme cannot meet the growing demand of our country.

Method used

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  • Preparation method for separating lysozyme by amino-functionalized magnetic nanoparticles
  • Preparation method for separating lysozyme by amino-functionalized magnetic nanoparticles
  • Preparation method for separating lysozyme by amino-functionalized magnetic nanoparticles

Examples

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Effect test

Embodiment 1

[0021] 3g FeCl 2 ·4H 2 O and 8.1g FeCl 3 ·6H 2 O was dissolved in 200 mL of deionized water, at room temperature, under nitrogen protection, 85 mL of 25% ammonia water was added dropwise to it, stirred for 30 min, then heated to 70 °C and continued to stir for 30 min, cooled to room temperature, collected with a magnet, and washed with deionized water for several times Rinse and dry to get magnetic Fe 3 O 4 Nanoparticles. Put 4g of magnetic Fe 3 O 4 The nanoparticles were placed in 50 mL of anhydrous toluene and subjected to ultrasound for 15 min. Under nitrogen protection, 22 nmol of 3-aminopropyltriethoxysilane (APTES) was added and stirred at room temperature for 5 h. After repeated rinsing and drying, amino-functionalized magnetic nanoparticles can be obtained.

[0022] Take the egg white of fresh eggs and phosphate buffer solution (20nmol) and dilute them at a volume ratio of 1:1, and then continuously stir the reaction in an ice-water bath for 6 hours. Insoluble...

Embodiment 2

[0025] 3g FeCl 2 ·4H 2 O and 8.1g FeCl 3 ·6H 2 O was dissolved in 200 mL of deionized water, at room temperature, under nitrogen protection, 85 mL of 25% ammonia water was added dropwise to it, stirred for 30 min, then heated to 70 °C and continued to stir for 30 min, cooled to room temperature, collected with a magnet, and washed with deionized water for several times Rinse and dry to get magnetic Fe 3 O 4 Nanoparticles. Put 4g of magnetic Fe 3 O 4 The nanoparticles were placed in 50 mL of anhydrous toluene and subjected to ultrasound for 15 min. Under nitrogen protection, 22 nmol of 3-aminopropyltriethoxysilane (APTES) was added and stirred at room temperature for 5 h. After repeated rinsing and drying, amino-functionalized magnetic nanoparticles can be obtained.

[0026] Take the egg white of fresh eggs and phosphate buffer solution (20nmol) and dilute them at a volume ratio of 1:1, and then continuously stir the reaction in an ice-water bath for 6 hours. Insoluble...

Embodiment 3

[0029] 3gFeCl 2 4H 2 O and 8.1 g FeCl 3 ·6H 2 O was dissolved in 200mL of deionized water, at room temperature, under the protection of nitrogen, 85mL of 25% ammonia water was added dropwise to it, stirred for 30min, then heated to 70°C and continued to stir for 30min, cooled to room temperature, collected with a magnet, and washed with deionized water several times Rinse and dry to obtain magnetic Fe 3 o 4 Nanoparticles. 4g magnetic Fe 3 o 4 Nanoparticles were put into 50mL of anhydrous toluene, sonicated for 15min, under the protection of nitrogen, 22nmol 3-aminopropyltriethoxysilane (APTES) was added, stirred at room temperature for 5h, magnetic particles were collected with a magnet, and the magnetic particles were collected with ethanol and dichloromethane After repeated washing and drying, amino functionalized magnetic nanoparticles can be obtained.

[0030] Take the egg white of fresh eggs and phosphate buffer solution (20nmol) to dilute at a volume ratio of 1:1...

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Abstract

The invention discloses a preparation method of separating lysozyme by amino-functionalized magnetic nanoparticles and specifically designs a method for separating lysozyme from egg white by the use of a magnetic nanoparticles adsorption material. According to the preparation method, Fe3O4 nanoparticles are selected and undergo surface-amino functionization modification so as to obtain amino-functionalized magnetic nanoparticles; the nanoparticles are added into an egg white solution and are saturated after 1 h of adsorption at 37 DEG C; the amino-functionalized magnetic nanoparticles saturated by adsorption are added into a phosphate buffer (20 nmol, pH=8.0, 1 mol/L NaCl) to be eluted at 37 DEG C for 2 h; and magnetic separation is carried out to obtain a solution, the eluate is put into a refrigerator of 4 DEG C, and centrifugation is carried out to obtain a solid enzyme powder. The separation method for extraction of egg white lysozyme is simple and has high separation efficiency. The obtained lysozyme has high purity. In addition, conditions are mild and have little influence on lysozyme activity, and recovery rate is high.

Description

technical field [0001] The invention designs a preparation method for separating lysozyme from amino-functionalized magnetic nano-particles, and particularly designs a method for separating lysozyme from egg white by utilizing a magnetic nano-adsorption material, which belongs to the technical field of fine chemicals. Background technique [0002] Lysozyme, also known as muramidase, its chemical name is N-acetyl muramidase, composed of a polypeptide chain, cross-linked by 4 disulfide bonds in the molecule, very stable in structure, and resistant to acid and alkali. It can still maintain good stability in the range of pH 1.2-11.2, and it is also extremely stable to heat. Under the condition of pH 3.0, it still maintains more than 90% in boiling water bath for 60min. In 1907, Nicolle first published a report on Bacillus subtilis bacteriolytic factor. Two years later, Laschtschenk pointed out that eggs also have strong bacteriolytic activity, and named it lysozyme. After Flemi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/30B01J20/22B01J20/28C12N9/36
Inventor 牛效迪周欣蕊王虹苏吕卓刁梦雪
Owner JILIN UNIV
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