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Preparation method and application of macroporous penicillin acylase immobilized magnetic bead polymer carrier

A technology of penicillin acylase and polymer, which is applied in the direction of immobilization on/in an organic carrier, can solve the problems of lack of functional groups, low synthesis cost, and many preparation procedures in magnetic microspheres, and achieve favorable Industrial application, low synthesis cost and simple process

Inactive Publication Date: 2013-01-16
NINGXIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Analysis of the above preparation process and experiments shows that this invention has some shortcomings: (1) Secondary initiation of polymerization is adopted, and the preparation procedures are many and the process is long
(2) The prepared magnetic microspheres lack specific functional groups, which is not conducive to the application in biomedicine, especially in the fields of immobilized enzymes and cell separation
[0006] At present, there is no synthetic method with low cost, simple process and no need for Fe 3 o 4 The preparation and surface modification procedures, the preparation method of macroporous penicillin acylase immobilized magnetic bead polymer carrier which is beneficial to industrial application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 0.66 g FeCl 2 4H 2 O and 1.82 g FeCl 3 ·6H 2O was placed in 15 mL of formamide, heated at 60 °C for 10 min with vigorous stirring, then the temperature was raised to 90 °C, ammonia water was added to adjust the pH to 8–9, and the magnetic fluid was formed after aging for 1 h. Add 40 mL of tetrachlorethylene and 110 mL of n-heptane into a reactor equipped with a constant speed stirrer, nitrogen tube and condenser, and after heating to 60 °C in a water bath, add 0.20 g of Span- 60 and 9.0 g calcium stearate, then add 2.8 g glycidyl methacrylate, 3.8 g N, N′-methylenebisacrylamide and the magnetic fluid prepared above, stir for 30 min and then add 1.0 g azobis Isobutyronitrile was controlled to rotate at 450 r / min, and polymerized under nitrogen protection for 7 h. The product was separated in a magnetic field, washed with acetone and n-heptane, and dried in vacuum to constant weight. The average pore diameter was measured to be 32.1 nm, the pore volume was 0.53 cm3 / g,...

Embodiment 2

[0028] 1.32 g FeCl 2 4H 2 O and 1.82 g FeCl 3 ·6H 2 O, the consumption and preparation condition of all the other components are identical with embodiment 1. The average pore diameter of the synthesized sample is 30.0 nm, the pore volume is 0.60 cm3 / g, the s value is 8.3 emu / g, and the epoxy group content is 0.38 mmol / g. The penicillin acylase was immobilized on a synthetic magnetic bead-like carrier containing an epoxy group, and was used to catalyze the hydrolysis reaction of penicillin G potassium to prepare 6-APA. The apparent activity of the immobilized enzyme was 612 IU / g.

Embodiment 3

[0030] Formamide was 30 mL, and the amounts and preparation conditions of the remaining components were the same as in Example 1. The average pore diameter of the synthesized sample is 39.4 nm, the pore volume is 0.51 cm3 / g, the s value is 6.3 emu / g, and the epoxy group content is 0.43 mmol / g. Penicillin acylase was immobilized on a synthetic magnetic bead-like carrier containing epoxy groups to catalyze the hydrolysis reaction of penicillin G potassium to prepare 6-APA. The apparent activity of the immobilized enzyme was 662 IU / g.

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Abstract

The invention relates to a preparation method of a polymer carrier, particularly a preparation method of a macroporous penicillin acylase immobilized magnetic bead polymer carrier, which comprises the following steps: dissolving Fe<2+> and Fe<3+> salts used as magnetic source precursors in a polar organic solvent formamide while introducing nitrogen, regulating the pH value of the solution to 8-11 with concentrated ammonia water, curing at 60-90 DEG C for 30-60 minutes, and forming a magnetic fluid under alkaline conditions; by using a surfactant as a dispersant, a vinyl compound as a monomer and a nonpolar organic substance as a dispersion medium, carrying out cross-linking polymerization under the action of an initiator by an reversed phase suspension technique; and washing with acetone to separate the product in a magnetic field, washing with an organic solvent, and carrying out vacuum drying at room temperature to obtain the carrier. The invention has the advantages of unique method, low synthesis cost and simple process, omits the processes of Fe3O4 preparation and surface modification, is beneficial to industrialized application, and can obtain the magnetic macroporous bead penicillin acylase immobilized carrier. The carrier is used for preparing high-activity segregative immobilized enzymes by penicillin acylase immobilization.

Description

technical field [0001] The invention relates to a preparation method of a polymer carrier, in particular to a preparation method of a macroporous penicillin acylase-immobilized magnetic bead polymer carrier and its application. Background technique [0002] Penicillin acylase, or PGA, is an industrial enzyme for the production of semi-synthetic β-lactam antibiotics. Its important application is to catalyze the key intermediate 6-aminopenicillane required for the production of semi-synthetic β-lactam antibiotics from penicillin G. The acid is 6-APA. At present, nearly 90% of 6-APA in the world is produced by penicillin acylase biocatalyst immobilized on natural polymer or synthetic resin carrier. Although immobilized PGA has been used in the pharmaceutical industry for nearly three decades, the preparation of new immobilized PGA carriers suitable for industrial applications is still challenging. The preparation of immobilized enzyme by magnetic carrier, the separation, wash...

Claims

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Application Information

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IPC IPC(8): C08F222/38C08F220/32C08F226/10C08F220/28C08F2/32C08F2/44C08K3/16C08K3/30C12N11/08
Inventor 薛屏李新燕夏维涛王居兰宋肖单李鹏
Owner NINGXIA UNIVERSITY
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