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Magnetic/functionalized SiO2 composite microsphere immobilized enzyme and preparation method thereof

A technology of composite microspheres and silica, which is applied in the direction of microsphere preparation, immobilization on or in inorganic carriers, microcapsule preparations, etc., can solve the problems of difficult recycling and reuse, and achieve improved operational stability, good biological Compatibility, the effect of easy preparation method

Inactive Publication Date: 2009-12-30
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although water-soluble enzymes have many advantages that other catalysts cannot match, because water-soluble enzymes are produced according to the needs of organisms themselves, they are greatly limited in practical application. The main disadvantages are: after the reaction, it is difficult to recycle and reuse
So far, there has been no report of using it in the immobilization research of biological enzymes

Method used

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  • Magnetic/functionalized SiO2 composite microsphere immobilized enzyme and preparation method thereof
  • Magnetic/functionalized SiO2 composite microsphere immobilized enzyme and preparation method thereof
  • Magnetic/functionalized SiO2 composite microsphere immobilized enzyme and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 5.40g FeCl 3 ·6H 2 O, 200mL of ethylene glycol and 14.40g of sodium acetate were stirred evenly, then transferred to a crystallization kettle, and placed in a 200°C oven for 8 hours, then cooled to room temperature, washed with ethanol for 6 times, and dried at 60°C to obtain magnetic microspheres. Then take 0.20g and use 100mL 0.1M hydrochloric acid to sonicate, then wash 3 times with water, then ultrasonically disperse into the mixed system of 160mL ethanol, 37mL water and 5mL ammonia water (25%), add 0.36mL tetraethyl orthosilicate and 0.04mLγ -Aminopropyltriethoxysilane (V 硅烷化试剂 / (V 硅烷化试剂 +V 正硅酸 乙酯)=10%), reacted for 6h, and magnetically separated and washed several times to obtain magnetic / aminated silica composite microspheres. Add 1.60mL glutaraldehyde solution to 0.10g magnetic / aminated silica composite microspheres to activate for 2h, recover by magnetic separation and wash with deionized water for several times, then add 1.20mL penicillin acylase at a cer...

Embodiment 2

[0022] 5.40g FeCl 3 ·6H 2 O, 200mL of ethylene glycol and 14.40g of sodium acetate were stirred evenly, then transferred to a crystallization kettle, and placed in a 200°C oven for 8 hours, then cooled to room temperature, washed with ethanol for 6 times, and dried at 60°C to obtain magnetic microspheres. Then take 0.20g and use 100mL 0.1M hydrochloric acid to sonicate, then wash 3 times with water, then ultrasonically disperse into the mixed system of 160mL ethanol, 37mL water and 5mL ammonia water (25%), add 0.36mL tetraethyl orthosilicate and 0.04mLγ -Aminopropyltriethoxysilane (V 硅烷化试剂 / (V 硅烷化试剂 +V 正硅酸 乙酯 )=10%), reacted for 6h, and magnetically separated and washed several times to obtain magnetic / aminated silica composite microspheres. Add 1.20mL of a certain concentration of penicillin acylase diluent to 0.10g of magnetic / aminated silica composite microspheres, vibrate and fix in a water bath shaker at 30°C for 24h, magnetically separate and wash with phosphate buf...

Embodiment 3

[0024] Same as Example 1, keep other conditions constant, change the adding volume of ethyl orthosilicate and γ-aminopropyltriethoxysilane into 0.20mL and 0.20mL respectively in the composite microsphere preparation process, i.e. V 硅烷化试剂 / (V 硅 烷化试剂 +V 正硅酸乙酯 ) = 50%.

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Abstract

The invention relates to a magnetic / functionalized SiO2 composite microsphere immobilized enzyme and a preparation method thereof, wherein, magnetic microspheres are cores, functionalized SiO2 is the shell, and the type and the content of functionalized groups, the particle size of the core in composite microsphere and the thickness of functionalized SiO2 can all be adjusted. The preparation method comprises the following steps: first synthesizing superparamagnetic microspheres, then treating the microspheres with hydrochloric acid, dispersing the microspheres in an ethanol / water alkaline system evenly by ultrasound, using tetraethoxysilane and silylating agent for further condensation and coating to obtain magnetic / functionalized SiO2 composite microspheres and performing absorbent immobilized and covalent immobilization to biological enzyme. The immobilized enzyme has fast magnetic responsiveness and good operational stability. The method has simple process and low cost and is applicable to industrialized production.

Description

technical field [0001] The invention relates to a magnetic / functionalized silicon dioxide composite microsphere immobilized enzyme and a preparation method thereof. More specifically, it is to prepare a magnetic / functionalized silica composite microsphere immobilized enzyme that can be rapidly magnetically separated and has good operational stability. Background technique [0002] Enzyme is a biological catalyst whose chemical essence is protein. Due to its high catalytic activity, strong selectivity, and mild reaction conditions, enzymes have been widely used in many fields such as biochemical industry, pharmaceutical chemical industry, food industry, and light industry. At present, thousands of enzymes have been discovered. Although water-soluble enzymes have many advantages that other catalysts cannot match, they are greatly limited in practical application because they are produced according to the needs of organisms themselves. The main disadvantage is that it is diff...

Claims

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Application Information

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IPC IPC(8): C12N11/14B01J13/02
Inventor 王艳芹石变芳卢冠忠王筠松郭杨龙任家文刘晓晖郭耘张志刚
Owner EAST CHINA UNIV OF SCI & TECH
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