Co-crosslinking immobilization method of penicillin G acylase

Abstract

The invention discloses a co-crosslinking immobilization method of penicillin G acylase. According to the co-crosslinking immobilization method of the penicillin G acylase, oil-soluble butanediol diacrylate is used as a crosslinking agent; penicillin G acylase containing amino groups, as well as a supramolecular complex formed by aminated epoxy resin and beta-cyclodextrin are used as reactants inan aqueous phase; and then, through a Michael addition reaction of the amino groups with double bonds, co-crosslinking polymerization is conducted at a relatively low temperature so as to prepare immobilized penicillin G acylase with different loading capacity. With crosslinking degree controlled, dispersivity enhanced and internal mass transfer microenvironment improved, the immobilized penicillin G acylase has relatively high catalytic activity that relative activity of the immobilized penicillin G acylase can be up to 91% of that of free enzyme when the loading capacity is 37 milligrams ofthe immobilized penicillin G acylase per gram of carrier.

Description

technical field [0001] The invention relates to the technical field of immobilized enzyme biocatalysis, in particular to a method for co-crosslinking and immobilizing penicillin G acylase. The novel immobilized penicillin G acylase can be specially used as a specific catalyst for various reactions. Background technique [0002] Penicillin G acylase (EC 3.5.1.11), also known as penicillin aminohydrolase, belongs to the class of aminohydrolase, which can specifically hydrolyze amide bonds, and is an important industrial enzyme. Penicillin G acylase belongs to globulin with larger molecular weight, and its three-dimensional structure consists of a large subunit containing a catalytic site and a small subunit containing a side chain binding site (by hydrogen bonding. The isoelectric point of the enzyme is about In 6.5~6.7. Penicillin G acylase is composed of α and β subunits. When α or β subunit exists alone, it has no enzymatic activity, and it is only active when the two are c...

AI Technical Summary

Problems solved by technology

The most commonly used cross-linking agent is water-soluble glutaraldehyde, which has high reactivity and is difficult to control the dosage. It is easy to cause excessive cross-linking of the enzyme and cause a great loss of enzyme activity. In addition, the traditional cross-linking method often It is necessary to precipitate and aggregate enzyme molecules before cross-linking, which will not only cause waste of enzymes, but also block mass transfer channels, and cannot fully exert the catalytic efficiency of enzymes

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