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Anionic type laminated material immobilized enzyme and its preparation method

A technology for immobilizing enzymes and layered materials, which can be applied in the direction of immobilization on or in inorganic carriers, immobilization on/in organic carriers, etc. Problems such as small load

Inactive Publication Date: 2003-12-03
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because enzymes are easily combined with organic polymers in a chemical way, it is more and more widely used, but the main problems are: polymer carriers have poor hydrophilicity, small enzyme loading, and low chemical stability.
Its disadvantage is that due to the strong acidity of the α-zirconium phosphate laminate, this method of directly immobilizing enzymes is only suitable for a small amount of enzymes that can survive in this pH range, and because the enzymes interact with the carrier through ionic bonds , the force is relatively weak, making the thermal stability of the immobilized enzyme poor, which is almost the same as that of the free enzyme
The operational stability of immobilized enzymes is not discussed in the literature

Method used

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  • Anionic type laminated material immobilized enzyme and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A: Preparation of Mg 2+ : Al 3+ = 2:1 Mg-Al hydrotalcite, take 4.0g and add it to a container containing 100ml deionized water, stir it magnetically at room temperature, add 7.5g of excess glutamic acid, at this time, the container changes from a cloudy solution to a clear solution ,spare.

[0026] Prepare a NaOH solution with a concentration of 0.05M, pour it into a reactor with a condensing and stirring device, and add the above-mentioned standby clear solution dropwise while stirring, keeping the pH of the dropping process > 9. After the dropping is completed, condense and reflux for 6 hours. Filtrate, wash fully with hot water until pH<8, and dry at 85°C for 24 hours to obtain a Mg-Al type hydrotalcite-like layer column material with glutamic acid as the interlayer anion.

[0027] B: Take the above 0.5g of glutamic acid pillared LDHs in a beaker filled with 30ml of deionized water, add 10ml of 5% glutaraldehyde, stir for 1 hour with magnetic force, filter, and ful...

Embodiment 2

[0032] Preparation of Mg 2+ : Al 3+ =3:1 Mg-Al hydrotalcite, get 4.0g and join in the container that fills 100ml deionized water, magnetically stir at room temperature, add excess glutamic acid 8.0g, prepare by the method for embodiment 1 and obtain interlayer anion It is a Mg-Al type hydrotalcite-like layer column material of glutamic acid.

[0033] B: The immobilized penicillin acylase was prepared according to the method of Example 1.

[0034] The X-ray diffraction pattern and IR characterization results of the sample prepared in step B are similar to Example 1, which proves that the penicillin acylase molecule has been successfully immobilized between the laminates.

[0035] The operational stability study of the immobilized enzyme showed that after 60 continuous operations in the batch reactor, the immobilized enzyme activity maintained 80% of its initial activity; the thermostability study of the immobilized enzyme showed that when the free enzyme activity was at 45 T...

Embodiment 3

[0037] A: Preparation of Mg 2+ : Al 3+ =4: 1 Mg-Al hydrotalcite, get 4.0g and join in the container that fills 100ml deionized water, stir magnetically at room temperature, add excess glutamic acid 9.0g, prepare by the method for embodiment 1 and obtain interlayer The anion is the Mg-Al type hydrotalcite-like layer column material of glutamic acid.

[0038] B: The immobilized penicillin acylase was prepared according to the method of Example 1.

[0039] The results of X-ray diffraction and IR characterization of the sample prepared in step B are similar to Example 1, which proves that the penicillin acylase molecule has been successfully immobilized between the laminates.

[0040] The operational stability study of the immobilized enzyme showed that after 40 continuous operations in the batch reactor, the immobilized enzyme activity maintained 80% of its initial activity; the thermostability study of the immobilized enzyme showed that when the free enzyme activity was at 45 ...

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Abstract

A anionic laminated material immobilized enzyme is prepared through intercalating the moleculae of object containing carboxy and amino groups between the layers of quasi-hydrotalcite as anionic laminated material, adding cross-linking agent and enzyme, and forming covalent bond between the amino groups on object and enzyme moleculae under the action of cross-linking agent to immobilize the enzyme moleculae between layers of quasi-hydrotalcite, so obtain layer column. Its advantage is high stability of enzyme moleculae.

Description

[0001] Field: [0002] The invention relates to an anionic layer material immobilized enzyme and a preparation method thereof. Background technique: [0003] As a green catalyst, enzymes have many advantages such as high catalytic efficiency, wide catalytic reaction range, high catalytic specificity, high affinity with substrates, mild catalytic reaction conditions, low energy consumption, and regeneration. Therefore, enzymes are used more and more in pharmaceutical and chemical industries, especially heavy chemical industries. Since free enzymes (FreeEnzyme) generally exist in aqueous solution at a relatively low concentration, their survival temperature and pH range are relatively narrow, they are easily denatured and inactivated, and their stability is poor, which greatly limits their application. [0004] In order to overcome the shortcomings of free enzymes, people thought of choosing an appropriate carrier to immobilize the enzyme, so that the thermal and chemical stabi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02C12N11/08
CPCC12N11/02C12N11/08
Inventor 段雪任玲玲何静
Owner BEIJING UNIV OF CHEM TECH