Metal-organic framework immobilized β-glucosidase and its preparation method and application

A metal-organic framework and glucosidase technology, applied in the direction of immobilized enzymes, biochemical equipment and methods, fixed on/in organic carriers, etc., can solve the problems of enzyme catalytic activity, weak binding force of surface adsorption method, To solve the problems of high preparation cost, achieve the effect of improving recycling rate, optimizing immobilization conditions and improving stability

Active Publication Date: 2021-08-03
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the immobilization of enzymes on MOFs materials mainly includes four methods: covalent binding, surface adsorption, pore embedding and co-precipitation. Catalytic activity; the surface adsorption method has weak binding force and is easy to desorb; the pore embedding method requires high reaction conditions and high preparation costs. In view of the simple process and mild conditions of the co-precipitation method, the co-precipitation method is generally used to immobilize the enzyme

Method used

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  • Metal-organic framework immobilized β-glucosidase and its preparation method and application
  • Metal-organic framework immobilized β-glucosidase and its preparation method and application
  • Metal-organic framework immobilized β-glucosidase and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Preparation of metal organic framework material (Cu(PABA)) and its immobilized enzyme β-G@Cu(PABA)

[0064] Weigh 50 µL of β-glucosidase, mix it with 50 µL of polyvinyl pyrrolidone (PVP) solution, mix it with 50 mM copper acetate solution, and then add 12.5 mM PABA solution to fully mix. Incubate statically in a constant temperature shaking incubator at 25°C for 8 hours, centrifuge at 4°C after the reaction is completed, and store the product in a refrigerator at 4°C after repeated washing for 3 times, and obtain metal-organic framework materials (Cu(PABA)) and It immobilizes the enzyme β-G@Cu(PABA).

Embodiment 2

[0065] Example 2: Characterization of metal-organic framework material (Cu(PABA)) and its immobilized enzyme β-G@Cu(PABA)

[0066] (1) Analysis of Fourier Transform Infrared Spectroscopy (FTIR)

[0067] The molecular structures of free β-glucosidase, metal-organic framework material Cu(PABA) and immobilized enzyme β-G@Cu(PABA) were detected by Fourier transform infrared spectrometer. First, KBr was dried overnight in an oven at 90 °C, and the treated sample and KBr were mixed at an equivalent ratio of 1:100, ground and pressed into tablets. Taking a separate KBr thin section as a blank, measure the infrared spectrograms of these three samples respectively, and record the infrared spectrograms of these three samples, such as figure 2 shown.

[0068] The metal-organic framework material (Cu(PABA)) and the immobilized enzyme β-G@Cu(PABA) were successfully synthesized by examining the positions of the characteristic group absorption peaks in the infrared spectrum and their shif...

Embodiment 3

[0081] Embodiment 3: The immobilization condition optimization of β-glucosidase

[0082] (1) Effect of metal ion and organic acid concentration on immobilization

[0083] The premise of the synthesis of immobilized enzyme β-G@Cu(PABA) is to be able to form the metal-organic framework material Cu(PABA). The corresponding concentration of copper acetate and p-aminobenzoic acid (PABA), as shown in Table 1 and Table 2, when the concentration of PABA is 6.25 mM, with the increase of copper acetate concentration, β-G@Cu(PABA) The immobilized amount and retained enzyme activity of the acid were slightly increased, because when the acid concentration was low, the amount of β-G@Cu(PABA) produced was less, and the free enzyme was introduced into the skeleton channel less. When the concentration of PABA was 12.5 mM, both immobilized capacity and retained enzyme activity were greatly increased, and when Cu 2+ When the concentration reaches 50 mM, the maximum immobilization capacity of β...

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Abstract

The invention relates to a metal organic framework material immobilized β-glucosidase and its preparation method and application; it belongs to the technical field of material preparation; the invention adopts a co-precipitation method to introduce a β-glucosidase into metal copper ions and p-aminobenzoic acid to form In the pore structure of the metal organic framework material Cu(PABA), the immobilization of the enzyme is realized; the metal organic framework material of the present invention is grown according to the enzyme, that is, the immobilization of the enzyme and the preparation of the framework material are carried out simultaneously; The preparation of the immobilized material and the immobilization of the enzyme are completed in one step, the stability of the enzyme, the resistance to organic solvents and metal ions are improved, the recycling rate is significantly improved, and the efficiency of cellulase to hydrolyze cellulose is also improved.

Description

technical field [0001] The invention belongs to the technical field of material preparation, and in particular relates to a metal-organic framework material immobilized beta-glucosidase and a preparation method and application thereof. Background technique [0002] At present, the demand for renewable energy has shifted from biodiesel to bioethanol produced from lignocellulose. The first step in bioethanol production is to hydrolyze cellulose to produce glucose. Compared with acid catalysis, enzyme catalysis has certain advantages. Such as eliminating the hindrance of side reactions to the fermentation process, compared with inorganic catalysis, enzyme catalysis is mild and efficient. Enzyme catalyzed hydrolysis of cellulose is composed of endo- and exo-glucanase and β-glucoside of cellulase. The enzymes cooperate to complete, firstly, cellobiose is produced by endo- and exoglucanase through a series of hydrolysis, and finally β-glucosidase hydrolyzes cellobiose to produce g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/082C12N9/42C12P19/14C12P19/02
CPCC12N9/2445C12N11/08C12P19/02C12P19/14C12Y302/01021
Inventor 王赟韩娟毛艳丽王蕾李春梅倪良敖文梅
Owner JIANGSU UNIV
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