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Enzyme-lectin conjugate nano-particles and preparing method thereof

A nanoparticle and lectin technology, applied in the direction of biochemical equipment and methods, enzymes, enzymes, etc., can solve the problems of inability to construct efficient multi-enzyme-magnetic nanoparticle systems, enzyme molecule inactivation, etc., to achieve easy industrial implementation and Scale-up, mild reaction conditions, and easy reusability

Active Publication Date: 2017-08-11
北京德润天勤生物工程技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Magnetic nanoparticle carrier is a commonly used carrier in nanobiocatalysis. However, it is easy to inactivate enzyme molecules and other characteristics, so it is impossible to construct an efficient multi-enzyme-magnetic nanoparticle system.

Method used

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  • Enzyme-lectin conjugate nano-particles and preparing method thereof
  • Enzyme-lectin conjugate nano-particles and preparing method thereof
  • Enzyme-lectin conjugate nano-particles and preparing method thereof

Examples

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Effect test

Embodiment 1

[0060] Embodiment 1, preparation enzyme-lectin conjugate nanoparticle

[0061] 1. Raw material ratio

[0062] In parts by weight, the proportioning of each raw material in the present embodiment is as follows:

[0063] 2 parts of glucose oxidase, 8 parts of horseradish peroxidase, 110 parts of concanavalin A (lectin), 0.4 parts of glutaraldehyde (crosslinking agent), 0.4 parts of sodium borohydride (reducing agent).

[0064] 2. Preparation method

[0065] Prepare the enzyme-lectin conjugate nanoparticles according to the following steps with the raw materials of the above ratio:

[0066] 1) Add enzyme and lectin to 10mM phosphate buffer solution with pH 7 (add 1mL buffer solution for every 1mg lectin), and stir (250rad / min) at 25°C for 2 hours;

[0067] 2) Add a cross-linking agent to the system obtained in step 1), continue to react for 2 hours, add a reducing agent after the cross-linking reaction, and continue to react for 24 hours;

[0068] 3) The system obtained in st...

Embodiment 2

[0073] Embodiment 2, preparation enzyme-lectin conjugate nanoparticle

[0074] The operation steps are the same as in Example 1, except that 0.4 parts of glutaraldehyde crosslinking agent is replaced by 300 parts of terminal aldehyde group PEG (i.e. aldehyde group-polyethylene glycol-aldehyde group, CHO-PEG-CHO) (Mn= 2000), the photos of the high-resolution transmission electron microscope of the prepared enzyme-lectin conjugate nanoparticles have no substantial difference from Example 1, and the biocatalytic activity is 61% of the equivalent natural enzyme, such as figure 2 shown.

Embodiment 3

[0075] Example 3, preparation of enzyme-lectin conjugate nanoparticles

[0076] The operation steps are the same as in Example 1, except that 0.4 parts of glutaraldehyde crosslinking agent is replaced by 200 parts of terminal succinimide ester PEG (succinimide ester-polyethylene glycol-succinimide ester, NHS -PEG-NHS) (Mn=2000), without using a reducing agent, the high-resolution transmission electron microscope photos of the prepared enzyme-lectin conjugate nanoparticles have no substantial difference from Example 1, and its biocatalytic activity is equivalent to natural 63% of enzymes, such as figure 2 shown.

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Abstract

The invention discloses enzyme-lectin conjugate nano-particles and a preparing method thereof. According to the preparing method, raw materials for preparing the enzyme-lectin conjugate nano-particles include enzyme, lectin and a cross-linking agent, as well as magnetic nano-particles and / or a reductant. The enzyme-lectin conjugate nano-particles can be prepared from, by weight, 10 parts of enzyme, 50-500 parts of lectin, 0.1-300 parts of cross-linking agent and 0-1 parts of reductant. The enzyme-lectin conjugate nano-particles can also be prepared from, by weight, 10 parts of enzyme, 50-500 parts of lectin, 1-100 parts of magnetic nano-particles, 0.1-300 parts of cross-linking agent and 0-1 parts of reductant. The enzyme-lectin conjugate nano-particles are dispersed in aqueous solution at a conventional catalytic reaction temperature and have high catalytic activity. Lectin combined with enzyme can attract saccharides substrates, so that the aqueous-phase activity of the conjugate can be further improved. Enzyme molecules in enzyme-lectin-magnetic nano-particle conjugate nano-particles are adsorbed by lectin, covalent cross-linking occurs between enzyme molecules and lectin and magnetic nano-particles, and therefore recoverability of the catalyst is realized based on the realization of high catalytic activity.

Description

technical field [0001] The invention relates to an enzyme-lectin conjugate nanoparticle and a preparation method thereof, belonging to the technical field of immobilized enzymes. Background technique [0002] As a kind of biocatalyst, enzyme has been widely used in the fields of medicine, food processing and fine chemical synthesis because of its high selectivity and mild reaction conditions. The biggest disadvantage of the free enzyme is its instability, which is prone to denaturation in acid, alkali, heat and organic solvents, and the activity is reduced or lost; and after the enzyme reaction, it will remain in the solution, making it difficult to continue and automate the enzyme reaction. It is also not conducive to the separation and purification of the final product, which greatly hinders the development of the enzyme industry. [0003] Nanobiocatalytic systems refer to systems based on the integration of enzymes and nanostructures, such as nanosilicon, polymer nanogel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/08C12N11/02
CPCC12N9/0006C12N9/0065C12N11/02C12N11/08C12N11/14C12Y101/03004C12Y111/01007
Inventor 雍有张一飞王瑞戈钧刘铮
Owner 北京德润天勤生物工程技术有限公司
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