Primary amine immobilized enzyme carrier and preparation method thereof

A technology of immobilizing enzyme carrier and primary amine group, applied in the direction of immobilization on/in organic carrier, fermentation, etc., to achieve the effects of high mechanical strength, low monomer price and simple process operation

Active Publication Date: 2018-01-30
天津南开和成科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems of the above-mentioned existing immobilized enzyme carriers, the present invention provides a novel carrier with high chemical stability, high enzyme activity and mechanical strength and a preparation method thereof. Macroporous polymethacrylate-diallylamine polymer, good storage stability, high mechanical strength and for penicillin G acylase, D-amino acid oxidase, GL-7-ACA acylase, etc. A variety of industrial enzymes have good immobilization effect

Method used

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  • Primary amine immobilized enzyme carrier and preparation method thereof
  • Primary amine immobilized enzyme carrier and preparation method thereof
  • Primary amine immobilized enzyme carrier and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Polymerization reaction:

[0027] i. with the AIBN of 1 mass part, the MA of 40 mass parts, the MMA of 20 mass parts, the EGDMA of 35 mass parts, the DAA of 5 mass parts, the toluene of 75 mass parts, the liquid wax of 75 mass parts, stir and mix, Formulated as an oil phase.

[0028] ii. Add 7.5 parts by mass of hydroxyethyl cellulose to 750 parts by mass of water, and stir to prepare an aqueous phase.

[0029] iii. Mix the oil phase with the water phase, stir to a suitable particle size range, heat up to 80°C, filter out the liquid after reacting for 15 hours, wash the solid with hot water and acetone, and sieve the 60-120 mesh carrier to obtain a crude carrier.

[0030] (2) Amination reaction:

[0031] The crude carrier was mixed with EDA, and reacted under stirring at 75°C for 18 hours. After the reaction was completed, the liquid was filtered out, and the carrier was washed with water until neutral to obtain a finished product.

[0032] (3) The appearance of ...

Embodiment 2

[0039] (1) Polymerization reaction:

[0040] i. the AIBN of 0.5 mass parts, the MA of 30 mass parts, the MMA of 10 mass parts, the EGDMA of 55 mass parts, the DAA of 5 mass parts, the toluene of 40 mass parts, the liquid wax of 60 mass parts, stir and mix, Formulated as an oil phase.

[0041] ii. Add 9 parts by mass of hydroxyethyl cellulose to 600 parts by mass of water, stir to prepare an aqueous phase.

[0042] iii. Mix the oil phase with the water phase, stir to a suitable particle size range, heat up to 70°C, filter out the liquid after reacting for 18 hours, wash the solid with hot water and acetone, and sieve a 30-60 mesh carrier to obtain a crude carrier.

[0043] (2) Amination reaction:

[0044] The crude carrier was mixed with TETA, and reacted under stirring at 75°C for 24 hours. After the reaction was completed, the liquid was filtered out, and the carrier was washed with water until neutral to obtain a finished product.

[0045] (3) The appearance of the immobi...

Embodiment 3

[0052] (1) Polymerization reaction:

[0053] i. the AIBN of 1.2 mass parts, the MA of 30 mass parts, the MMA of 10 mass parts, the EGDMA of 35 mass parts, the DAA of 25 mass parts, the toluene of 133 mass parts, the liquid wax of 67 mass parts, stir and mix, Formulated as an oil phase.

[0054] ii. Add 7.5 parts by mass of hydroxyethyl cellulose to 900 parts by mass of water, and stir to prepare an aqueous phase.

[0055] iii. Mix the oil phase with the water phase, stir to a suitable particle size range, heat up to 85°C, filter out the liquid after 12 hours of reaction, wash the solid with hot water and acetone, and sieve the 60-120mesh carrier to obtain a crude carrier.

[0056] (2) Amination reaction:

[0057] The crude carrier was mixed with TDA, and reacted under stirring at 75°C for 14 hours. After the reaction was completed, the liquid was filtered out, and the carrier was washed with water until neutral to obtain a finished product.

[0058] (3) The appearance of th...

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Abstract

The invention discloses a primary amine immobilized enzyme carrier and a preparation method thereof. Positive suspension polarization is performed to obtain a crude macroporous polymethacrylate-diallylamine carrier; washing the crude carrier, and grading; performing amination to obtain the primary amine-containing immobilized enzyme carrier. The primary amine immobilized enzyme carrier according to the technical scheme of the invention is applicable to immobilization of various biological enzymes, provides effective immobilization for penicillin G acylase, D-amino acid oxidase, Gl-7-ACA acylase and the like, and has high enzyme-carrying activity, good mechanical strength and good storage stability. The preparation method is simple and has low cost, and the carrier has significantly improved storage stability and is expectedly applicable to industrial large-scale production.

Description

technical field [0001] The invention belongs to the field of biochemical industry and relates to the preparation of a carrier for enzyme immobilization, in particular to a method for preparing an immobilized enzyme carrier with a primary amine group. Background technique [0002] Biological enzymes have the characteristics of high efficiency, specificity, mild reaction conditions, and environmental protection. There are about 5,000 enzymes that have been discovered so far, and there are more than 60 enzymes that can be industrially produced. There are more than 20 major industrial enzymes. In terms of development direction, on the one hand, the industrial and scientific communities are still constantly searching for new strains, developing strain transformation and propagation techniques, and researching and developing new enzyme preparation application processes; on the other hand, how to use enzymes more effectively and rationally, both It has become an important issue to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12P37/06C12P37/00C12P35/02C08F251/02C08F220/14C08F222/14C08F8/32
Inventor 于浩峰崔强李晓蒙王鹏陈利马苓王红红郑彦慧杨晶
Owner 天津南开和成科技有限公司
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