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Gel immobilization enzyme with catalytic and antibacterial functions and preparation method thereof

A technology of immobilizing enzymes and gels, which is applied to the direction of immobilization on or in inorganic carriers, or in or on organic carriers, which can solve the problems of limited application, difficult recycling, and easy contamination of bacteria, etc., and achieve the goal of overcoming Poor mechanical stability, easy to use continuously, and the effect of solving the problem of bacterial contamination

Inactive Publication Date: 2018-10-12
天津市农业科学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cross-linked enzyme aggregates have high enzyme activity per unit and are highly resistant to high temperature, organic solvents, acid-base, metal ions, etc., but the mechanical strength of cross-linked enzyme aggregates is poor and the particle size is small (usually micron) , It is not easy to recycle, and the appearance is an amorphous structure, which limits its application in industry
In addition, when immobilized enzymes such as cross-linked enzyme aggregates are used continuously in the solution for a long time, because the catalytic reaction conditions are usually suitable for microbial growth, bacteria are easily contaminated during the reaction process, which seriously affects the catalytic reaction effect

Method used

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  • Gel immobilization enzyme with catalytic and antibacterial functions and preparation method thereof
  • Gel immobilization enzyme with catalytic and antibacterial functions and preparation method thereof

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preparation example Construction

[0017] The preparation of cross-linked enzyme aggregates can refer to [Schoevaart R, Wolbers M W, GolubovicM, Ottens M, Kieboom A P, van Rantwijk F, van der Wielen L A, Sheldon R A. Preparation, optimization, and structures of cross-linked enzyme aggregates (CLEAs). Biotechnology and Bioengineering, 2004, 87(6):754-762.]. Among them, enzymes include but are not limited to protease, penicillin acylase, urease, pectinase, lactase, trehalose synthase, glucose isomerase, glycosidase, amylase, pullulanase, glucose oxidase, peroxidase Hydrogenase, laccase, phytase, organophosphate hydrolase, etc. or a combination thereof.

[0018] Polymers capable of forming gels include, but are not limited to, gelatin, pectin, agar, polyvinyl alcohol, sodium alginate, carrageenan, konjac gum, chitosan, dextran, polyacrylamide, silica gel, and the like. Only one kind of polymer can be used, or two or more kinds of polymers can be blended for use. The polymer is pre-dissolved before use. For exam...

Embodiment 1

[0024] In this embodiment, the enzyme used is trehalose synthase, the polymer used is chitosan, and the nanoparticles with antibacterial function used are nano Ag.

[0025] Use 50mmol / L, pH7.0 phosphate buffer to prepare 5mL trehalose synthase solution, the protein content of which is 5mg / mL, add glucose to the enzyme solution to make the final concentration of glucose 2% (w / v), gradually add 1.2g of PEG 6000, after the PEG is dissolved, add glutaraldehyde, the final concentration of glutaraldehyde is 0.2% (v / v), mix well, then cross-link at 25°C for 2h, then centrifuge at 3000r / min for 20min, The resulting precipitate was centrifuged and washed three times with 10 mmol / L, pH 7.0 phosphate buffer solution to obtain cross-linked enzyme aggregates of trehalose synthase, which were suspended in 5 mL of water for later use.

[0026] Add silver nitrate solution to the chitosan solution prepared by acetic acid solution, the final concentration of chitosan is 1% (w / v), the final conc...

Embodiment 2

[0029] The preparation method is the same as in Example 1, except that 0.02 g of Fe is added to the resulting 4 mL mixture. 3 o 4 Magnetic powder, the resulting gel-immobilized enzyme can be separated and recovered by using an external magnetic field.

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Abstract

The invention discloses a gel immobilization enzyme with catalytic and antibacterial functions and a preparation method thereof. The gel immobilization enzyme has not only the catalytic reaction activity of enzymes, but also the antibacterial property, so that microbiological contamination is not prone to happen in an enzymic catalytic reaction process; moreover, the gel immobilization enzyme is easy to recover, so that reuse is convenient.

Description

technical field [0001] The invention relates to a gel-immobilized enzyme with both catalytic and antibacterial functions and a preparation method thereof, belonging to the field of enzyme immobilization. Background technique [0002] Compared with free enzymes, immobilized enzymes have the advantages of high stability, easy separation from the reaction system, reusability, and the realization of automated production. The rapid development of the biocatalysis industry is of great significance. Cross-linked enzyme aggregate is a carrier-free immobilized enzyme. Its preparation principle is: add salt, organic solvent, non-ionic polymer and other precipitants to the enzyme solution, so that the enzyme protein can form a supramolecular structure through non-covalent bonds. - Insoluble physical aggregates, and then use cross-linking agents to covalently bind the aggregates to obtain cross-linked enzyme aggregates. Cross-linked enzyme aggregates have high enzyme activity per unit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10C12N11/04
CPCC12N11/04C12N11/10C12N11/14
Inventor 张峻陈颖杨丽维赵尉吏
Owner 天津市农业科学院
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