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Mesoporous titanium dioxide immobilized enzyme and preparation method and application thereof

A technology of mesoporous titanium dioxide and immobilized enzymes, which is applied in the fields of immobilization on or in inorganic carriers, chemical industry, sustainable manufacturing/processing, etc. It can solve the problems of unreported immobilization methods and achieve good reusability , Applicable pH and temperature range expansion, the effect of structural stability

Inactive Publication Date: 2010-07-14
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the application of γ-glutamyl transpeptidase all uses the free enzyme as the catalyst, and there is no report on its immobilization method and its properties.

Method used

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  • Mesoporous titanium dioxide immobilized enzyme and preparation method and application thereof
  • Mesoporous titanium dioxide immobilized enzyme and preparation method and application thereof
  • Mesoporous titanium dioxide immobilized enzyme and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] M-TiO 2 Balance of carrier: weigh 5g M-TiO 2 For the mesoporous material, the carrier was equilibrated with 250 mL of Tris-HCl buffer solution of pH 8.050 mmol / L for 24 hours, and stored in the above buffer solution at 4° C. for future use.

[0033] Mesoporous material M-TiO 2 Immobilization of γ-glutamyl transpeptidase on the carrier: the above M-TiO 2 Drain, weigh 1gM-TiO 2 Put the wet granules in a sand core tube, add 10 mL of 0.038 mg / mL γ-glutamyl transpeptidase solution (enzyme activity is 22.35 U / mL), and shake and react in an ice bath at 4°C for 2.5 hours. After the reaction, wash off the excess enzyme solution with a Tris-HCl buffer solution of pH 8.050mmol / L, and store it in the above buffer solution to obtain M-TiO 2Immobilized γ-glutamyl transpeptidase. Its enzyme activity is 131U / g carrier, and the recovery rate of enzyme activity is 58.6%.

Embodiment 2

[0035] M-TiO 2 Balance of carrier: weigh 5g M-TiO 2 For the mesoporous material, the carrier was equilibrated with 250 mL of Tris-HCl buffer solution of pH 8.050 mmol / L for 24 hours, and stored in the above buffer solution at 4° C. for future use.

[0036] Mesoporous material M-TiO 2 Immobilization of γ-glutamyl transpeptidase on the carrier: the above M-TiO 2 Drain, weigh 1gM-TiO 2 The wet granules were placed in a sand core tube, 10 mL of 0.073 mg / mL γ-glutamyl transpeptidase solution (enzyme activity: 22.62 U / mL) was added, and the reaction was shaken in an ice bath at 4°C for 4 hours. After the reaction, wash off the excess enzyme solution with a Tris-HCl buffer solution of pH 8.050mmol / L, and store it in the above buffer solution to obtain M-TiO 2 Immobilized γ-glutamyl transpeptidase. Its enzyme activity is 119U / g carrier, and the recovery rate of enzyme activity is 52.6%.

Embodiment 3

[0038] M-TiO 2 Balance of carrier: weigh 5g M-TiO 2 For the mesoporous material, the carrier was equilibrated with 250 mL of Tris-HCl buffer solution of pH 8.050 mmol / L for 24 hours, and stored in the above buffer solution at 4° C. for future use.

[0039] Mesoporous material M-TiO 2 Immobilization of γ-glutamyl transpeptidase on the carrier: the above M-TiO 2 Drain, weigh 1gM-TiO 2 Put the wet granules in a sand core tube, add 10 mL of 0.017 mg / mL γ-glutamyl transpeptidase solution (enzyme activity is 21.07 U / mL), and shake and react in an ice bath at 4°C for 2.5 hours. After the reaction, wash off the excess enzyme solution with a Tris-HCl buffer solution of pH 8.050mmol / L, and store it in the above buffer solution to obtain M-TiO 2 Immobilized γ-glutamyl transpeptidase. Its enzyme activity is 107.8U / g carrier, and the recovery rate of enzyme activity is 51.15%.

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Abstract

The invention discloses a mesoporous titanium dioxide immobilized enzyme, which comprises enzyme and a carrier to immobilize the enzyme. The enzyme is a gamma- glutamyltranspeptidase, and the carrier is mesoporous titanium dioxide. The invention also discloses a preparation method of the mesoporous titanium dioxide immobilized enzyme and application thereof. The mesoporous titanium dioxide immobilized enzyme obtained through the invention has the advantages of homogeneous property, large surface area, high enzyme stability, good reusability and the like, and establishes foundations to further improve the stability of the gamma-glutamyltranspeptidase and expand the industrial application of the glutamyl acylation reaction.

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] The non-toxic and environmentally friendly titanium dioxide material is a new type of material that has attracted much attention in recent years. It has broad prospects as an ideal titanium-containing active carrier. Mesoporous titanium dioxide (M-TiO 2 ) has excellent characteristics such as large specific surface area, strong chemical stability, and good mechanical operation strength, so the development and production of M-TiO 2 Mesoporous materials and immobilized enzyme products based on the materials are very meaningful. [0003] γ-glutamyltranspeptidase (γ-glutamyltranspeptidase, GGT, EC 2.3.2.2) can specifically transfer γ-glutamyl to acceptor molecules to obtain new γ-glutamyl-containing compounds. become a glutamylation reaction. Different glutamyl compounds can be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12P13/02C12P21/02
CPCY02P20/50
Inventor 姚忠秦丹华王浩琦徐红韦萍
Owner NANJING UNIV OF TECH
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