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Immobilized enzyme using modified epoxy resin as carrier and preparation method and application thereof

An epoxy-based resin and immobilized enzyme technology, which is applied in the fields of immobilization on/in organic carriers, chemical industry, climate sustainability, etc., can solve the problem of immobilization and stabilization of γ-glutamyl transpeptidase There are few researches, immobilization and application have not been reported, and the expansion of application range is limited, so as to achieve the effect of good reusable performance, uniform shape, and increased applicable pH and temperature range.

Inactive Publication Date: 2012-07-11
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because GGT is prone to subunit depolymerization in vitro, it causes an irreversible decrease in enzyme activity, which limits the further expansion of its application range.
[0007] At present, there are few studies on the immobilization and stabilization of γ-glutamyl transpeptidase, and there are no reports on the immobilization and application of GGT using ampholyte-modified materials as carriers.

Method used

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  • Immobilized enzyme using modified epoxy resin as carrier and preparation method and application thereof
  • Immobilized enzyme using modified epoxy resin as carrier and preparation method and application thereof
  • Immobilized enzyme using modified epoxy resin as carrier and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Preparation of ECCA carrier.

[0045] Take 0.3g dried Eupergit C 250L Put it in a 20ml sand core tube, add 50% (v / v) CA aqueous solution according to the solid-to-liquid ratio of 1g / 20mL, and react at room temperature for 16h. After the reaction is completed, the reaction liquid will be filtered out first. The dried solid particles were rinsed repeatedly with pure water for 6 to 7 times; then rinsed with 0.5mol / L NaCl solution for 3 to 4 times; finally rinsed with pure water for 6 to 7 times and then filtered dry. The CA-modified carrier ECCA was obtained by drying at 40°C. The grafted amount of CA calculated by the mass difference method was 0.0964g.

Embodiment 2

[0046] Example 2: Preparation of ECCA carrier.

[0047] Weigh 0.3g of dried Eupergit C 250L Put it in a 20ml sand core tube, add 100% (v / v) CA according to the solid-to-liquid ratio of 1g / 10mL, and react at room temperature at 24°C for 20h. After the reaction, the reaction liquid will be filtered out first. The dried solid particles were rinsed repeatedly with pure water for 6 to 7 times; then rinsed with 0.5mol / L NaCl solution for 3 to 4 times; finally rinsed with pure water for 6 to 7 times and then filtered dry. The CA-modified carrier ECCA was obtained by drying at 40°C. The grafted amount of CA calculated by mass subtraction method was 0.124g.

Embodiment 3

[0048] Example 3: Preparation method of ECCA-GGT.

[0049] Take by weighing 0.3g of the dry product carrier ECCA prepared by the most preferred method in Example 2 and put it in the ampoule bottle, add the purified GGT enzyme solution (enzyme concentration is 0.3mg / mL) according to the solid-liquid ratio 1g: 13.3mL, and in Shake the reaction in a water bath at room temperature for 1 hour. After the reaction, the reaction liquid is filtered out, and the filtered solid particles are repeatedly rinsed with pure water for 3 to 4 times, and dried at 30°C to obtain ECCA-GGT. The adsorption capacity of GGT was 1.26mg / g resin, and the recovery rate of enzyme activity was 9.72%.

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Abstract

The invention discloses an immobilized enzyme using modified epoxy resin as a carrier, which comprises an enzyme and the modified carrier for fixing the enzyme. The enzyme is gamma-glutamyl transpeptidase, and the modified carrier is the epoxy resin Eupergit C250L modified through carrier ampholyte with potential of hydrogen (pH) 8-10.5. The invention further discloses a preparation method and application of the immobilized enzyme. The immobilized enzyme has the advantages of being uniform in shape, strong in enzyme stability, good in repeated use performance, strong in reaction environment pH adaptability and the like. Stability of the gamma-glutamyl transpeptidase is improved further, and application of the novel immobilized enzyme in synthesis of theanine is achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an immobilized enzyme and its preparation method and application. Background technique [0002] Biocatalytic reactions have the characteristics of mild conditions, high selectivity, and high reaction efficiency, so they are increasingly used in the field of chemical synthesis. However, the poor stability of biological enzymes in in vitro industrial environments and the high cost of enzyme preparations greatly limit the practical application of biocatalysis in the industrial field. Immobilized enzyme technology not only effectively solves the problem of catalyst reuse by binding (or binding) enzyme molecules to a special phase, but also significantly improves the stability and catalytic properties (substrate affinity, stereoselectivity, etc.) of biological enzymes. ). [0003] At present, as an important branch of immobilized enzyme technology, the development and applic...

Claims

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Application Information

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IPC IPC(8): C12N11/08C12P13/04
CPCY02P20/50
Inventor 姚忠杨敬王浩绮周治房鑫徐虹韦萍
Owner NANJING UNIV OF TECH
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