Method for preparing immobilized glucose isomerase with composite magnetic chitosan microballoon spheres

A technology of glucose isomerase and composite microspheres, which is applied in the field of preparation of immobilized glucose isomerase by magnetic chitosan composite microspheres, can solve the problems of non-reusable use, waste, and low efficiency of glucose isomerase use, and achieve The effect of easy operation, lower production cost and increased yield

Inactive Publication Date: 2010-06-23
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] An object of the present invention is to provide a method for preparing immobilized glucose isomerase with magnetic chitosan composi

Method used

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  • Method for preparing immobilized glucose isomerase with composite magnetic chitosan microballoon spheres
  • Method for preparing immobilized glucose isomerase with composite magnetic chitosan microballoon spheres

Examples

Experimental program
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Embodiment 1

[0018] 1. To 0.4mol / L FeCl 2 100ml and FeCl 3 In 100ml, add 10.0g of polyglycol with a molecular weight of 4000, fully dissolve it under a magnetic stirrer, adjust the pH of the solution to 10 with ammonia water, continue to stir for 30min, wash with double distilled water and vacuum freeze-dry to obtain Fe 3 o 4 magnetic core.

[0019] 2. Dissolve chitosan powder in 3% acetic acid and disperse ultrasonically for 10 minutes to prepare a 0.02g / ml chitosan solution. 3 o 4 Mix with magnetic nuclear ultrasound and electric stirring for 10 minutes to form a microemulsion system, and cross-link with 5% glutaraldehyde, continue stirring for 4 hours at 2000r / min, then wash with petroleum ether, acetone and double-distilled water, and vacuum After freeze-drying, the magnetic chitosan composite microspheres are obtained. The emulsifier consists of 60ml liquid paraffin, 1ml span-80, 1ml Tween-60, and 1ml n-butanol.

[0020] 3. Soak the prepared magnetic chitosan composite microsphe...

Embodiment 2

[0022] 1. To 0.5mol / L FeCl 2 30ml and FeCl 3 Add 6.0g of polyethylene glycol with a molecular weight of 4000 to 50ml, fully dissolve it under a magnetic stirrer, adjust the pH of the solution to 8 with ammonia water, continue to stir for 50min, wash with double distilled water and vacuum freeze-dry to obtain Fe 3 o 4 magnetic core.

[0023] 2. Dissolve the chitosan powder in 6% acetic acid for 10 minutes of ultrasonic dispersion to make a 0.08g / ml chitosan solution, and pass the emulsifier and Fe 3 o 4 Ultrasonic disperse the magnetic nucleus and mix with electric stirring for 10 minutes to form a microemulsion system, and cross-link with 1% glutaraldehyde, continue stirring for 2 hours at 2000r / min, then wash with petroleum ether, acetone and double distilled water, and filter with suction The magnetic chitosan composite microspheres are obtained after vacuum freeze-drying.

[0024] 3. Soak the prepared magnetic chitosan composite microspheres with phosphate buffer pH 7....

Embodiment 3

[0026] 1. Using 0.6mol / L FeCl 2 30ml and FeCl 3 50ml, add 8.0g of polyethylene glycol with a molecular weight of 4000, fully dissolve under a magnetic stirrer, adjust the pH of the solution to 9 with ammonia water, continue to stir for 30min, wash with double distilled water and vacuum freeze-dry to obtain Fe 3 o 4 magnetic core.

[0027] 2. Dissolve the chitosan powder in 5% acetic acid for 10 minutes of ultrasonic dispersion to make a 0.05g / ml chitosan solution, and pass the emulsifier and Fe 3 o 4 Mix with magnetic nuclear ultrasound and electric stirring for 10 minutes to form a microemulsion system, and cross-link with 2% glutaraldehyde, continue stirring for 2 hours at 2000r / min, then wash with petroleum ether, acetone and double-distilled water, filter and vacuum After freeze-drying, the magnetic chitosan composite microspheres are obtained. Wherein the emulsifier consists of 80ml liquid paraffin, 2.5ml span-80, 1.5ml Tween-60, and 2ml n-butanol.

[0028] 3. Soak ...

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Abstract

The invention relates to a method for preparing immobilized glucose isomerase with composite magnetic chitosan microballoon spheres, which comprises the steps of: adding polyethylene glycol into FeCl2 and FeCl3, dissolving, regulating pH of solution to 8 to 10 with ammonia water, washing, filtering, freezing and drying in vacuum to obtain Fe3O4 magnetic nuclei, dissolving chitosan powder in acetic acid to carry out ultrasonic dispersion to prepare chitosan solution, carrying out ultrasonic dispersion and stirring on emulsifying agents and the Fe3O4 magnetic nuclei, crosslinking 1 to 5% glutaric dialdehyde at 2000 r/min, stirring, washing, filtering, freezing and drying in vacuum to obtain composite magnetic chitosan microballoon spheres, soaking the composite magnetic chitosan microballoon spheres with phosphate buffer pH of which is 7.0 to 7.8, filtering, adding glucose isomerase diluted by buffer, vibrating, taking out the glucose isomerase, putting the glucose isomerase in a refrigerator for one night, pouring off supernate, precipitating, washing, and filtering to obtain the immobilized glucose isomerase. In the invention, the glucose isomerase activity recovery ratio of which can reach more than 80% is immobilized by the composite magnetic chitosan, and has high service efficiency, reusability and low cost.

Description

1. Technical field: [0001] The invention relates to a method for preparing immobilized glucose isomerase by magnetic chitosan composite microspheres. 2. Background technology: [0002] Glucose isomerase (GI) is mainly an intracellular enzyme, also known as xylose isomerase, which can convert aldose into corresponding ketose, that is, convert glucose into fructose, thereby preparing fructose syrup. However, during the use of free enzymes, not only the use efficiency is low, but also they cannot be reused, resulting in great waste. At present, there are also many methods for immobilizing glucose isomerase, but some methods have disadvantages such as weak binding ability between the enzyme and the carrier, and easy detachment and loss. 3. Contents of the invention: [0003] An object of the present invention is to provide a method for preparing immobilized glucose isomerase from magnetic chitosan composite microspheres, which is used to solve the problem of low use efficienc...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10
Inventor 曹龙奎王菲菲张学娟
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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