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Modified epoxy resin immobilized enzyme and preparation method and application thereof

An epoxy resin, immobilized enzyme technology, applied in the field of enzyme catalysis, can solve the problems of low recovery rate of enzyme activity, weak action force, singleness, etc.

Active Publication Date: 2021-09-07
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, commercial epoxy resins usually only have a single functional group, and the interaction with enzymes is weak; resulting in low enzyme immobilization efficiency, low recovery of enzyme activity, and poor reusability

Method used

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  • Modified epoxy resin immobilized enzyme and preparation method and application thereof
  • Modified epoxy resin immobilized enzyme and preparation method and application thereof
  • Modified epoxy resin immobilized enzyme and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] The preparation method of embodiment 1 modified epoxy resin immobilized inositol-1-phosphate synthase

[0098] Synthesize modified epoxy resin immobilized inositol-1-phosphate synthase according to the following steps:

[0099] Take 4g of epoxy resin 1000EP in a 50mL centrifuge tube, add 15mL of 50mM Tris-HCl buffer and stir for 15 minutes to maintain the pH of the overall system environment = 7.2. Suction filter after 1 hour, and wash the carrier with deionized water.

[0100] Weigh 1.0g of activated epoxy resin 1000EP into different 10mL centrifuge tubes, add 6mL of 5% ethylenediamine aqueous solution (pH=9.0) and place it on a rotary shaker for 2 hours. Use milli-Q to wash repeatedly to remove unreacted Ethylenediamine, wash by suction filtration; then add 2mL 0.25% glutaraldehyde to activate on a shaker for 1h, wash with buffer 3 times, then weigh 400mg of resin and place it in 1mL of 70% by mass ammonium sulfate precipitant solution (containing 0.472g of ammonium ...

Embodiment 2

[0108] Embodiment 2 Preparation method of immobilized inositol-1-phosphate synthase

[0109] Synthesize immobilized inositol-1-phosphate synthase according to the following steps:

[0110] Take 5g of epoxy resin 1000EP in a 50mL centrifuge tube, add 20mL of 50mM Tris-HCl buffer solution and stir for 15 minutes to maintain the pH of the overall system environment = 7.2, suction filter after 1 hour, and wash the carrier with deionized water.

[0111] Weigh 1.0g of activated epoxy resin 1000EP into different 10mL centrifuge tubes, add 6mL of 5% ethylenediamine aqueous solution (pH=9.0) and place it on a rotary shaker for 2 hours. Use milli-Q to repeatedly wash away unreacted ethylenediamine , wash by suction filtration; add 2mL of 0.25% glutaraldehyde to activate on a shaking table for 1h, wash 3 times with buffer, then weigh 400mg of resin and place it in 1mL of 70% by mass ammonium sulfate precipitant solution (containing 0.472 g ammonium sulfate), then add 1 mL of IPS enzyme ...

Embodiment 3

[0114] Embodiment 3 Preparation method of immobilized inositol-1-phosphate synthase

[0115] Synthesize immobilized inositol-1-phosphate synthase according to the following steps:

[0116] Take 8g of epoxy resin 1000EP in a 50mL centrifuge tube, add 32mL of 50mM Tris-HCl buffer and stir for 15 minutes to maintain pH = 7.2. Suction filter after 1 hour and wash the carrier with deionized water.

[0117] Weigh 4.0g of activated epoxy resin 1000EP into different 50mL centrifuge tubes, add 24mL of 5% ethylenediamine aqueous solution (pH=9.0) and place it on a rotary shaker for 2 hours, then use milli-Q to repeatedly filter and wash to remove unreacted ethylenediamine, then add 8mL 0.25% glutaraldehyde to activate on a shaking table for 1h, wash with buffer 3 times, then weigh 800mg modified epoxy resin and place it in 2mL, 70% by mass ammonium sulfate precipitant solution (containing 0.944g of ammonium sulfate), then add 2mL of IPS enzyme solution with a final concentration of 0.7...

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Abstract

The invention discloses a modified epoxy resin immobilized enzyme and a preparation method and application thereof, and belongs to the field of enzyme catalysis. The method comprises the steps that ethylenediamine is added into pretreated epoxy resin for modification, the modified epoxy resin is activated, then an enzyme to be immobilized is added for immobilization, the immobilized enzyme is obtained, and the epoxy resin with double functional groups is obtained. The phenomenon that the immobilization effect of one functional group is poor can be avoided, the enzyme activity of the immobilized enzyme is improved, the immobilization effect can be higher, the better reusability is achieved, and the immobilization time can be shortened. The preparation method is easy to operate and high in operability, and the prepared immobilized enzyme has the wide application prospect.

Description

technical field [0001] The invention belongs to the field of enzyme catalysis, and in particular relates to a modified epoxy resin-immobilized enzyme and its preparation method and application. Background technique [0002] Inositol, also known as cyclohexanhexol, is a water-soluble six-carbon ring vitamin B group compound. Chemical formula and glucose (C 6 h 12 o 6 ) same, but with a six carbon ring. Almost all cells contain inositol, which exists in free or bound form. Inositol is also a water-soluble vitamin that is necessary for human and animal growth. Widely used in cosmetics, feed, food and pharmaceutical industries. In the cosmetics industry, inositol can promote cell growth and prevent cell aging; in the feed industry, it can be used as a biological accelerator, which can promote the growth of livestock and prevent death; in the food industry, inositol is often used as a nutritional enhancer to prevent fat in humans Deposits in the body, especially cardiovasc...

Claims

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Application Information

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IPC IPC(8): C12N11/089C12N9/06C12P7/18
CPCC12N11/089C12N9/0022C12Y104/03007C12Y104/03011C12P7/18Y02P20/50
Inventor 崔建东王高杨杜英杰王梓源
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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