Method for preparing immobilized enzyme from zeolite imidazate skeleton compound

A skeleton compound, zeolite imidazolate technology, applied in the direction of fixing on or in the inorganic carrier, fixing on/in the organic carrier, etc., can solve the problems of unfavorable recovery and recycling, difficult separation and purification, poor stability, etc. Achieve the effect of cost-effective, simple operation and stable properties

Active Publication Date: 2020-01-24
JIANGSU UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the free carboxypeptidase A has poor stability during use, such as thermal stability, pH value tolerance range and tolerance to organic reagents, etc., and the free

Method used

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  • Method for preparing immobilized enzyme from zeolite imidazate skeleton compound
  • Method for preparing immobilized enzyme from zeolite imidazate skeleton compound
  • Method for preparing immobilized enzyme from zeolite imidazate skeleton compound

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1 zeolite imidazolate skeleton compound loaded carboxypeptidase A complex synthesis

[0029] (1) Weigh 0.0657g of 2-methylimidazole and add 5ml of ultrapure water to make 2-methylimidazole aqueous solution, take 0.439g of zinc acetate and add 5ml of ultrapure water to make zinc acetate aqueous solution, and make 1mL concentration 15mg / mL polyethylene glycol aqueous solution;

[0030] (2) Use a pipette gun to take 960 microliters (0.025g) of the enzyme stock solution, put it in a 1.5mL centrifuge tube and centrifuge for 5 minutes, remove the supernatant, and dissolve the 1mL volume of polyethylene glycol prepared in step (1) The aqueous solution is added to the centrifuge tube, and ultrasonicated for 15 minutes to form a polyethylene glycol / carboxypeptidase A complex solution of polyethylene glycol-coated carboxypeptidase A;

[0031] (3) Put the polyethylene glycol / carboxypeptidase A complex solution into an ultrafiltration centrifuge tube for ultrafiltration...

Embodiment 2

[0034] Example 2 Synthesis of zeolite imidazolate skeleton compound loaded carboxypeptidase A complex

[0035] (1) Weigh 1.0526g of 2-methylimidazole and add 8ml of ultrapure water to make 2-methylimidazole aqueous solution, take 0.1407g of zinc acetate and add 2ml of ultrapure water to make zinc acetate aqueous solution, and make 1mL concentration 15mg / mL polyethylene glycol aqueous solution;

[0036](2) Use a pipette gun to take 960 microliters (0.025g) of the enzyme stock solution, put it in a 1.5mL centrifuge tube and centrifuge (10000r / min) for 5 minutes, then remove the supernatant, and make the 1mL volume prepared in step (1) The polyethylene glycol aqueous solution is added in this centrifuge tube, ultrasonic 15 minutes forms the polyethylene glycol / carboxypeptidase A complex solution of polyethylene glycol encapsulation carboxypeptidase A;

[0037] (3) Put the polyethylene glycol / carboxypeptidase A complex solution into an ultrafiltration centrifuge tube for ultrafil...

Embodiment 3

[0040] Example 3 Synthesis of Carboxypeptidase A Complex Supported by Zeolite Imidazolate Skeleton Compound

[0041] (1) Weigh 1.5787g of 2-methylimidazole and add 8ml of ultrapure water to make 2-methylimidazole aqueous solution, take 0.1407g of zinc acetate and add 2ml of ultrapure water to make zinc acetate aqueous solution, and make 1mL concentration 15mg / mL polyethylene glycol aqueous solution;

[0042] (2) Use a pipette gun to take 960 microliters (0.025g) of the enzyme stock solution, put it in a 1.5mL centrifuge tube and centrifuge (10000r / min) for 5 minutes, then remove the supernatant, and make the 1mL volume prepared in step (1) The polyethylene glycol aqueous solution is added in this centrifuge tube, ultrasonic 15 minutes forms the polyethylene glycol / carboxypeptidase A complex solution of polyethylene glycol encapsulation carboxypeptidase A;

[0043] (3) Put the polyethylene glycol / carboxypeptidase A complex solution into an ultrafiltration centrifuge tube for u...

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Abstract

The invention discloses a method for preparing immobilized enzyme from a zeolite imidazate skeleton compound. The method comprises the following steps: respectively preparing a 2-methylimidazole solution, a zinc acetate aqueous solution and a polyethylene glycol aqueous solution; centrifuging an enzyme stock solution, removing a supernate, adding the polyethylene glycol aqueous solution, carryingout ultrasonic treatment to obtain a polyethylene glycol/carboxypeptidase A compound solution, carrying out ultrafiltration on the compound solution, and transferring the polyethylene glycol/carboxypeptidase A compound into the 2-methylimidazole solution by using the 2-methylimidazole solution to form a mixed solution; and adding the zinc acetate aqueous solution into the mixed solution, magnetically stirring, performing steps of standing and aging, centrifugally collecting a solid product, washing with water, and drying the material to obtain the immobilized enzyme prepared from the zeolite imidazate skeleton compound. According to the method, the stability and the loading rate of the enzyme can be greatly improved, the storage time of the enzyme is prolonged, and the prepared zeolite imidazate skeleton compound-loaded carboxypeptidase A compound is simple in recovery operation, economical, efficient and high in reusability.

Description

technical field [0001] The invention relates to immobilized enzyme technology, which is suitable for industrial fields such as grain food safety, and in particular relates to a method for preparing immobilized enzyme from a zeolite imidazolate skeleton compound. Background technique [0002] Enzyme immobilization methods include physical adsorption, coupling, interception, embedding, and cross-linking. Metal-organic framework compounds can be used as carrier materials for enzyme immobilization. Adsorb enzymes into the pores of pre-synthesized metal-organic framework compounds. Although this immobilization of enzymes can be achieved through simple steps, since the pore size of most metal-organic The method of forming complex enzymes has certain limitations. Even if the metal-organic framework compound has a pore size similar to or larger than that of the enzyme, if the binding between them is relatively weak, the enzyme is likely to be washed away, and the existing adsorptio...

Claims

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Application Information

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IPC IPC(8): C12N11/02C12N11/14
CPCC12N11/02C12N11/14
Inventor 孔德昭盛建国马蕾李波李雅琪康乐群裘茜
Owner JIANGSU UNIV OF SCI & TECH
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