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Renewable magnetic immobilized enzyme carrier and preparation method thereof

An immobilized enzyme carrier, magnetic technology, applied in the direction of immobilization on or in an inorganic carrier, etc., can solve the problems of increasing production cost, abandoning carrier, reducing activity, etc.

Inactive Publication Date: 2014-01-22
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The above magnetic nanocomposite carrier has significant advantages, but its shortcomings are also very obvious, such as the biocompatibility of these carriers needs to be further improved, and the biocompatibility of the carrier is directly related to the activity of immobilized enzymes; on the other hand, When the loaded enzyme undergoes multiple cycles of catalytic reactions, the activity will gradually decrease or even become inactive, which will eventually lead to the abandonment of these carriers, and the synthesis of new carriers will inevitably increase production costs.

Method used

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  • Renewable magnetic immobilized enzyme carrier and preparation method thereof
  • Renewable magnetic immobilized enzyme carrier and preparation method thereof
  • Renewable magnetic immobilized enzyme carrier and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: A kind of preparation method of regenerable magnetically immobilized enzyme carrier,

[0045] (a) Preparation of Fe 3 o 4 Magnetic nanoparticles (Fe 3 o 4 NPs):

[0046] Fe 3 o 4 Magnetic nanoparticles were synthesized by the following solvothermal method. Mix 3.24g FeCl under mechanical stirring 3 ·6H 2 O, 10.44g of sodium acetate, dissolved in 120mL of ethylene glycol solution; transfer the obtained yellow solution to a stainless steel reaction kettle lined with polytetrafluoroethylene, and react at 200°C for 12h; use an external magnetic field to separate and collect black magnetic Particles; the isolated product was washed with absolute ethanol for 4 times, then separated and collected, and placed at 60°C for complete volatilization for later use.

[0047] (b) Preparation of core-shell structure Fe 3 o 4 TiO 2 Magnetic colloidal nanoparticles (Fe 3 o 4 TiO 2 NPs):

[0048] 200 mg of Fe prepared in step (a) 3 o 4 NPs were added to a so...

Embodiment 2

[0059] Embodiment 2: a kind of preparation method of regenerable magnetically immobilized enzyme carrier,

[0060] Same as the steps of Example 1, the difference is that the step (e) of Example 1 is to connect the prepared carrier with horseradish peroxidase to detect the load of the carrier on the enzyme and the impact on the enzyme activity, while the embodiment The second is to link the carrier prepared according to steps (a) to (d) of Example 1 with laccase to detect the load of the carrier on laccase (EC1.10.3.2, Fluka) and the effect on enzyme activity. The results showed that Fe 3 o 4 TiO 2 The nanocomposite carrier loaded 98 ± 9 mg / g of laccase and retained 96 ± 4% of the activity of the free enzyme.

Embodiment 3

[0061] Embodiment 3: a kind of preparation method of regenerable magnetically immobilized enzyme carrier,

[0062] Same as the steps of Example 1, the difference is that the step (e) of Example 1 is to connect the prepared carrier with horseradish peroxidase to detect the load of the carrier on the enzyme and the impact on the enzyme activity, while the embodiment 3 is to connect the carrier prepared according to the steps (a) to (d) of Example 1 with alcohol dehydrogenase (EC1.1.1.1, Sigma) to detect the load of the carrier on the alcohol dehydrogenase and the effect on the enzyme activity . The results showed that Fe 3 o 4 TiO 2 The nanocomposite carrier loaded 105 ± 6 mg / g of alcohol dehydrogenase and retained 93 ± 5% of the activity of the free enzyme.

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Abstract

The invention discloses a renewable magnetic immobilized enzyme carrier and preparation method thereof, and belongs to the technical field of immobilized enzymes. The preparation method comprises the steps of compounding Fe3O4 nano-particles, preparing Fe3O4@TiO2 magnetic nano-composites of a core-shell structure, preparing anatase type Fe3O4@TiO2 nano-composites, carrying out functional modification on the surfaces of the nano-composites to obtain a Fe3O4@TiO2 immobilized enzyme carrier and the like. The immobilized enzyme carrier prepared through the preparation method composites the advantages of magnetic materials, nano materials, TiO2 and surficial functional group modification, has the advantages of having magnetic responsiveness, being renewable, high in biocompatibility, being in covalent link with target enzyme molecules, and being high in enzyme capacity value and good in stability. In addition, the preparation method is simple in technology, and reaction conditions are easy to control. According to the renewable magnetic immobilized enzyme carrier and the preparation method thereof, the immobilized carrier with excellent performance is provided for enzymes, cost of large-scale industrialized application of the enzymes can be reduced effectively, and the renewable magnetic immobilized enzyme carrier and the preparation method thereof have huge application potential in the field of immobilized enzymes.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzymes, in particular to a reproducible magnetic immobilized enzyme carrier and a preparation method thereof. Background technique [0002] Enzymes are a class of biocatalysts, and the chemical essence of most enzymes is protein. Compared with chemical catalysts, enzymes have the advantages of strong specificity, high catalytic efficiency, mild reaction conditions, and controllable activity. However, enzymes are unstable and are easily affected by the external environment and lose their catalytic activity. In addition, most enzymes are water-soluble, which makes it difficult to separate from the substrate and product after the catalytic reaction, which not only affects the purity of the product but also prevents the enzyme from being reused, which increases the cost of the enzyme in industrial and agricultural applications. [0003] The emergence of immobilized enzyme technology provides an...

Claims

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Application Information

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IPC IPC(8): C12N11/14
Inventor 蔡林君陈雷赵冰
Owner JILIN UNIV
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