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Compound immobilized enzyme carrier material and preparation method and application thereof

A technology for immobilizing enzyme carriers and carrier materials, which is applied in the directions of immobilization on/in organic carriers, biochemical equipment and methods, enzymes, etc. Application, not many functional groups, etc., to achieve the effects of good temperature and storage stability, rich surface functional groups, and high adsorption capacity

Active Publication Date: 2017-10-17
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies have found that although the pore size of most MOFs materials reported so far is adjustable, they are generally microporous structures (the pore size is about 2nm), so the size of the enzyme molecule plays a key role in the loading. Only enzyme molecules with small molecular diameters can be loaded, which limits the application of the interception method
The cross-linking method is to use a cross-linking agent (usually glutaraldehyde) to form a covalent bond with the functional group on the enzyme protein to immobilize the enzyme on the carrier. Although the cross-linking method makes the enzyme and the carrier more effective , the combination is stronger, but the enzyme activity will decrease
[0005] There are also a small number of studies using post-synthesis modification methods to make the surface of MOFs materials have functional groups that can react with enzymes to achieve the purpose of immobilizing enzymes, but the introduced functional groups are not many, resulting in only a small amount of enzymes passing through. Physical adsorption or covalent bonding on the surface of MOFs materials, the immobilization capacity of enzymes has not been greatly improved

Method used

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  • Compound immobilized enzyme carrier material and preparation method and application thereof
  • Compound immobilized enzyme carrier material and preparation method and application thereof
  • Compound immobilized enzyme carrier material and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Add 0.5g (0.285mmol) UiO-66-NH 2 Dissolve the solid in 20mL DMF, add 0.13mL (1.71mmol) 3-bromopropyne and 0.4g (2.91mmol) potassium carbonate, and react at 70°C for 24 hours under a nitrogen atmosphere. and absolute ethanol for 3 times, and vacuum-dried at room temperature to obtain the alkyne-modified UiO-66-NH 2 .

[0035] 2. Dissolve 0.11g (0.5mmol) 2-azidoethyl-2-bromo-methylpropyl ester, 7.1g (50mmol) tBMA and 0.0865g (0.5mmol) PMDETA in 5mL anhydrous methyl ethyl ketone and isopropanol In the mixed solution with a volume ratio of 1:1, add 0.0715g (0.5mmol) CuBr after two consecutive freeze-thaw degassing, freeze-thaw degassing once more, and then react at 60°C under a nitrogen atmosphere for 12 hours. After the reaction, Cool to room temperature, pass through a neutral alumina column after dilution with tetrahydrofuran, and remove most of the solvent from the eluate by a rotary evaporator, then precipitate in a mixed solvent with a volume ratio of methanol an...

Embodiment 2

[0039] The application of the compound immobilized enzyme carrier material of embodiment 1 in immobilizing pectinase, concrete method is as follows:

[0040] Add pectinase to citric acid-disodium hydrogen phosphate buffer solution with a pH value of 4.0 to prepare a 10 U / mL pectinase solution, then add 10 mg of composite immobilized enzyme carrier material to 25 mL of pectinase solution, and keep at room temperature React for 120 minutes, centrifuge at 4000 rpm for 5 minutes, and wash the precipitate with citric acid-disodium hydrogen phosphate buffer solution with pH=4.0 to obtain immobilized pectinase, which is stored at 4°C.

[0041] In order to determine the optimal process condition of the immobilized pectinase of composite immobilized enzyme carrier material of the present invention, inventor has carried out a large amount of laboratory research tests, and concrete test situation is as follows:

[0042] 1. The effect of immobilization time on the activity and adsorption ...

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Abstract

The invention discloses a compound immobilized enzyme carrier material and a preparation method and application thereof. The carrier material is prepared as follows: after alkynyl groups are introduced into a reaction of UiO-66-NH2 and 3-propargyl bromide, a click chemistry reaction occurs with poly(tert-butylmethacrylate) of which the end group is nitrine, and then hydrolyzing to obtain nano-particles with the grain diameter of 280 to 320 nm. Through electrostatic incorporation between carboxyl and amino, pectinase is fixed onto the carrier material, so that the active sites capable of being combined with pectinase are also greatly increased while the stability of the pectinase is improved, the adsorbing capacity of the pectinase is high and reaches 89.7% at most; the immobilized pectinase has higher pH, temperature and storage stability and preferable reusability, the relative enzyme activity after eight times of reusability is still 81%, and the carrier material has huge application prospect in the field of immobilized enzymes.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzymes, in particular to a composite immobilized enzyme carrier material and its preparation method and application. Background technique [0002] Immobilized enzymes have been widely used in many fields such as chemistry, medicine, and food because of their better temperature and pH stability, reusability, and easy separation of catalytic products than free enzymes. The choice of immobilized enzyme carrier material is an important factor to determine whether the enzyme can be immobilized successfully and the activity of the immobilized enzyme is high or low. An ideal carrier material should have good mechanical strength, chemical stability, thermal stability, high affinity for enzymes, and be able to maintain high enzyme activity. The carrier materials for immobilized enzymes have developed from the initial natural polymer materials to synthetic polymer materials, inorganic materials, and n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F120/18C08F8/32C12N11/08C12N9/88
CPCC08F8/32C12N9/88C12N11/08C12Y402/0201C08F120/18
Inventor 雷忠利李燕杨红
Owner SHAANXI NORMAL UNIV
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