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Magnetic enzyme nanogel biocatalytic particle and preparation method thereof

A magnetic nanoparticle and nanogel technology, applied in the direction of being fixed on or in an inorganic carrier, immobilized on/in an organic carrier, etc., can solve the problems of inability to recycle, poor thermal stability of natural enzymes, etc. No mass transfer diffusion resistance, reduce the loss of enzyme activity, prevent the effect of inactivation problems

Active Publication Date: 2012-07-25
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a magnetic enzyme nanogel particle to solve the problems of poor thermal stability and inability to recycle existing natural enzymes, which is of great significance for further developing the application of enzymes

Method used

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  • Magnetic enzyme nanogel biocatalytic particle and preparation method thereof
  • Magnetic enzyme nanogel biocatalytic particle and preparation method thereof
  • Magnetic enzyme nanogel biocatalytic particle and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of magnetic lipase nanogel biocatalytic particles

[0046] 1) Preparation of ferric oxide nanoparticles with superparamagnetism

[0047] Raw materials: 2 parts by mole of ferric chloride, 1 part by mole of ferrous chloride, 7 parts by weight of ammonia water (mass concentration 30%), 10 parts in total.

[0048] Dissolve ferric chloride and ferrous chloride in double-distilled water at a molar ratio of 2:1, add dilute hydrochloric acid dropwise to adjust the pH of the solution to less than 2, and pass nitrogen gas for 30 minutes to remove oxygen. Cool the above-mentioned ferric chloride / ferrous chloride solution to 4-8°C, and quickly add 7 parts of cooled ammonia water under the condition of rapid stirring (rotating speed greater than 1,000 rpm), to form a black colloidal solution. The temperature of the reaction system was increased to 70° C., and kept for 30 minutes, magnetically stirred, and nitrogen gas was kept flowing at the same time, to ob...

Embodiment 2

[0057] Example 2: Preparation of magnetic trypsin nanogel biocatalytic particles

[0058] Change the target enzyme in Example 1 into 20 parts of trypsin, and the magnetic particles into: 80 parts of ferric oxide nanoparticles II modified with carboxyl and double bonds prepared in Example 1, and 400 parts of vinyl-containing monomers Parts of acrylamide, 4 parts of methylene bisacrylamide as crosslinking agent, 8 parts of ammonium persulfate and 12 parts of N, N, N', N'-tetramethylethylenediamine as mixture of initiator. All the other steps are the same as in Example 1.

[0059] Finally, the trypsin magnetic enzyme nanogel was dispersed in the CaCl-containing 2 10mM, pH 8 in 50mM Tris-Cl activation buffer. Using Na-benzoyl-DL-arginine-p-nitroamide hydrochloride as a substrate to measure the biocatalytic activity of trypsin magnetic enzyme nanogel biocatalytic particles, the total yield was 95%, and the polymerization yield was 89%, and the product particle size is 200nm.

Embodiment 3

[0060] Example 3: Preparation of magnetic cytochrome acrylic nanogel biocatalytic particles

[0061] Change the target protein in Example 1 into 10 parts of cytochrome c, and the magnetic particles into: 40 parts of ferric oxide nanoparticles II modified with carboxyl and double bonds prepared in Example 1, and change the vinyl monomer into 300 parts Part of sodium acrylate and crosslinking agent were changed to 0.5 part of N, N'-methylenebisacrylamide, and the rest of the formula and steps were the same as in Example 1. At this point, the product obtained is measured with 2'-azino-bis(3-ethylbenzothiazole-6-sulfonic acid) and hydrogen peroxide as the total yield of the biocatalytic activity of the nanometer macromolecular biocatalytic particles. 82%, the polymerization yield is 70%, and the product particle size is 250nm.

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Abstract

The invention discloses a magnetic enzyme nanogel biocatalytic particle and a preparation method thereof. The magnetic enzyme nanogel biocatalytic particle is of a core-shell structure, the core of the magnetic enzyme nanogel biocatalytic particle is a magnetic nanoparticle with enzyme molecules adsorbed on the surface, and the enzyme molecules react with the magnetic nanoparticle through electrostatic force; the shell is a crosslinked high-polymer material layer; and the core is connected with the shell through a chemical bond. The preparation method is as follows: an amino group, a carbon-carbon double bond, a carboxyl group and other groups are first introduced into the surface of the magnetic nanoparticle by way of chemical modification, enzyme-particle nanoscale composite material is then obtained by regulating the pH of solution, and finally, with a vinyl monomer as material, the magnetic enzyme nanogel biocatalytic particle is obtained by way of free radical polymerization. The method is characterized in that: the method is simple and mild, purification is convenient, and industrial scale-up is easy to implement. The magnetic enzyme nanogel biocatalytic particle has the characteristics of high biocatalytic activity and high thermal stability, is convenient to recycle, and has a broad application prospect in organic synthesis, the food industry, the detergent industry, the energy industry, biomedicine, sensors and other fields.

Description

technical field [0001] The invention relates to a magnetic enzyme nano gel biocatalytic particle and a preparation method thereof. Background technique [0002] The stability and recyclability of natural enzymes are the main limiting factors for the industrial application of enzymes as biocatalysts. At present, chemical additive methods, immobilization methods, genetic engineering and other methods can improve the stability of enzyme molecules. However, additive methods need to add a large number of additives, and at the same time will bring new impurities and interference to the system; traditional immobilization methods introduce more High mass transfer resistance brings about a significant decrease in enzyme catalytic activity; while genetic engineering methods are more complex, costly, have limited effect on improving stability, and cannot solve the problem of mass and cheap production. At present, nanoscale catalytic particle preparations have attracted attention becau...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/08C12N11/04
Inventor 林萌萌刘铮卢滇楠
Owner TSINGHUA UNIV
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