Preparation of magnetic immobilized penicillin G acylase doped with divalent manganese ions

Abstract

The invention relates to preparation of magnetic immobilized penicillin G acylase doped with divalent manganese ions. The method comprises the following steps: (1) preparing magnetic nanoparticles ferroferric oxide@beta-cyclodextrin doped with divalent manganese ions; (2) carrying out mixed reaction on an ethanol solution of 3-glycidyl ether oxypropyl trimethoxy silane with the magnetic nanoparticle ferroferric oxide@beta-cyclodextrin doped with the divalent manganese ionst, so as to obtain a magnetic nanoparticle ferroferric oxide@beta-cyclodextrin-g-3-glycidyl ether oxypropyl trimethoxy silane carrier doped with the divalent manganese ions; and (3) adding the divalent manganese ion-doped magnetic nanoparticle ferroferric oxide@beta-cyclodextrin-g-3-glycidyl ether oxypropyl trimethoxy silane carrier dissolved in a phosphate buffer solution into a free penicillin G acylase solution, carrying out oscillatory reaction, and then carrying out vacuum drying to obtain the magnetic immobilized penicillin G acylase carrier doped with the divalent manganese ions. The method can effectively improve the enzyme activity recovery rate.

Description

technical field [0001] The invention relates to the technical field of preparation of functional materials, in particular to the preparation of magnetically immobilized penicillin G acylase doped with divalent manganese ions. Background technique [0002] At present, penicillin G acylase was first discovered in the 1960s and is an important biocatalyst with strong specificity and high catalytic efficiency. It can catalyze penicillin G potassium to produce 6-aminopenicillanic acid. Because 6-aminopenicillanic acid is a key intermediate of semi-synthetic penicillins, it occupies a huge market in the field of antibacterial and anti-inflammatory. [0003] Although penicillin G acylase has many advantages in the catalytic process, the special structural features make the conformation of free penicillin G acylase extremely unstable, prone to conformational changes and even the separation of the two subunits. Therefore, the direct use of free penicillin G acylase in the catalytic...

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Problems solved by technology

[0003] Although penicillin G acylase has many advantages in the catalytic process, the special structural features make the conformation of free penicillin G acylase extremely unstable, prone to conformational changes and even the separation of the two subunits

Therefore, the direct use of free penicillin G acylase in the catalytic process still has many disadvantages, such as poor environmental tolerance, easy to be affected by temperature, pH, etc., so that the enzyme activity is significantly reduced or even lost.

Moreover, after catalytic hydrolysis, penicillin G acylase is difficult to separate from the substrate and product, and the production process is difficult to achieve continuous operation, which will not only increase the difficulty of operation, prolong the process route, increase economic costs, but also cause pollution to the product

The above-mentioned problems of penicillin G acylase limit its industrial application in the production of 6-aminopenicillanic acid

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