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Immobilization method of free enzyme

A lipase and metal salt technology, applied in the field of immobilization of free enzymes, can solve problems such as poor structural stability, difficult separation of free enzymes, narrow particle size distribution, etc., achieve low mechanical loss rate, overcome crystal transformation, and maintain enzyme activity high rate effect

Pending Publication Date: 2021-09-17
WANHUA CHEM GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for immobilizing free enzymes. The method uses microemulsion suspension technology to adsorb the free enzymes on the modified inorganic non-metallic carrier in situ, effectively solving the problem that the free enzymes are not easy to separate during the catalytic process. , the problem of poor structural stability of traditional immobilized enzymes, the obtained immobilized enzymes have uniform nanometer size, narrow particle size distribution, and no agglomeration effect

Method used

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  • Immobilization method of free enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Two parts of identical microemulsions were prepared at 25°C, each part of microemulsion consisted of: 100g n-hexane / ethyl acetate (n-hexane 33.3g, ethyl acetate 66.7g), 100g dodecylaminopropionic acid and 100g deionized water.

[0031] (2) Weigh 10 g of CALB lipase dry powder, add 50 mL, 0.2 mol / L sodium hydrogen phosphate buffer solution therein, then add 10 g of barium chloride powder therein, and stir to obtain a white emulsion. The obtained white emulsion was added to one part of the microemulsion prepared in step (1), and it was used under stirring state, which was recorded as the microemulsion mixed liquid one.

[0032] (3) Prepare 100g mass fraction of 10wt% sodium sulfate aqueous solution, add it to the above-mentioned microemulsion two, then add 10g triethylamine therein, stir and mix it for later use, and record it as microemulsion mixed solution two.

[0033] (4) Add two drops of the microemulsion mixed solution in step (3) to the first microemulsion mix...

Embodiment 2

[0036] (1) Prepare two identical microemulsions at 25°C, each microemulsion consists of: 100g n-hexane / ethyl acetate (n-hexane 50.0g, ethyl acetate 50.0g), 50g dodecylaminopropionic acid and 100g deionized water.

[0037] (2) Weigh 10g of CALB lipase dry powder, add 60mL, 0.4mol / L sodium hydrogen phosphate buffer solution to it, then add 20g of barium chloride powder to it, and stir to obtain a white emulsion. The obtained white emulsion was added to one part of the microemulsion prepared in step (1), and it was used under stirring state, which was recorded as the microemulsion mixed liquid one.

[0038] (3) Prepare 100g mass fraction of 12wt% sodium sulfate aqueous solution, add it to the above-mentioned microemulsion two, then add 20g triethylamine therein, stir and mix it for later use, and record it as microemulsion mixed liquid two.

[0039] (4) Add two drops of the microemulsion mixed solution in step (3) to the first microemulsion mixed solution in step (2) at 15°C, an...

Embodiment 3

[0042] (1) Two identical microemulsions were prepared at 25° C., each microemulsion consisted of: 100 g of n-hexane, 100 g of dodecylalanine and 100 g of deionized water.

[0043] (2) Weigh 10 g of CALB lipase dry powder, add 50 mL, 0.2 mol / L sodium hydrogen phosphate buffer solution therein, then add 10 g of barium chloride powder therein, and stir to obtain a white emulsion. The obtained white emulsion was added to one part of the microemulsion prepared in step (1), and it was used under stirring state, which was recorded as the microemulsion mixed liquid one.

[0044] (3) Prepare 100 g of sodium sulfate aqueous solution with a mass fraction of 10 wt %, add it to the above-mentioned microemulsion two, then add 10 ethylenediamine therein, stir and mix it for later use, and record it as microemulsion mixed liquid two.

[0045] (4) Add two drops of the microemulsion mixed solution in step (3) to the first microemulsion mixed solution in step (2) at 10°C, control the dropping ti...

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Abstract

The invention discloses an immobilization method of a free enzyme. According to the method, a microemulsion suspension technology is adopted for the first time, the free enzyme is adsorbed on a covalently modified inorganic carrier in situ, and the defects that the free enzyme is not easy to separate in the catalytic process, the traditional immobilized enzyme is poor in structural stability, the unit utilization efficiency of the enzyme is low and the like are effectively overcome. The loading capacity of the obtained immobilized enzyme reaches 200mg / g, the size of the enzyme is nanoscale, the average diameter is 3.5-6.5nm, the dispersion is uniform, and the agglomeration phenomenon is avoided. In addition, the process does not need a harsh preparation environment, the enzyme activity retention rate can reach 100%, and the mechanical loss rate is less than 0.05%. The method is suitable for synthesizing enzymes for catalyzing macromolecules and micromolecules, and has universality.

Description

technical field [0001] The invention belongs to the technical field of chemical synthesis, and in particular relates to a method for immobilizing free enzymes. Background technique [0002] Immobilized enzyme technology refers to the enzyme preparation that is not easily soluble in water and has activity after being treated by physical or chemical methods. Compared with free enzymes, the advantages of immobilized enzymes are: first, effectively improve the catalytic performance and operational stability of enzymes, and reduce their costs. Second, immobilized enzymes are more conducive to the separation and purification of products than free enzymes, and show superior performance in terms of pH tolerance, substrate selectivity, thermal stability and recyclability. [0003] Traditional enzyme immobilization methods mainly include the following categories: adsorption method, chemical cross-linking method, embedding method and covalent binding method. The adsorption method mai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14
CPCC12N11/14
Inventor 罗朝辉
Owner WANHUA CHEM GRP CO LTD
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