Enzyme immobilization method based on excessive expenditure polymer

Abstract

The invention relates to an enzyme immobilization method based on a hyperbranched polymer, which belongs to the technical field of bioengineering. The method comprises: in a synthetic hyperbranched polymer solution with a large number of diazonium salt groups on the periphery, or In the dilute solution of the hyperbranched polymer solution diluted with water or organic solvent N,N-dimethylformamide less than 100 times, the carrier is soaked for more than half an hour; then the carrier is taken out, washed with water, and then immersed in In the enzyme-containing buffer solution, adsorb at 4°C for more than 4 hours to achieve saturated adsorption; this method can greatly simplify the operation steps of immobilized enzymes, without the need for pre-modification of materials and enzymes, and has high universality; it can be used On a variety of enzymes and carriers, and has no obvious effect on the catalytic activity of the enzyme.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular to a method for immobilizing enzymes and other bioactive components on various polymer films and inorganic materials. Background technique [0002] With the rapid development of biotechnology, bioengineering, as a frontier field of technological revolution, has been widely used in industry, agriculture, and medicine, and plays a decisive role in solving many problems faced by human beings today, such as resources, energy, and environmental protection. effect. [0003] As one of the components of bioengineering, enzyme engineering has been widely used in food, textile and other sectors in industrial production. However, due to the low stability of natural enzymes, they are extremely sensitive to high temperature and organic solvents, are easily inactivated, and cannot be reused. , the enzyme is mixed in the product after the catalyzed reaction, making it difficult to purify, ...

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Problems solved by technology

[0004] The commonly used enzyme immobilization methods are as follows: 1) non-covalent binding method, which can be divided into crystallization method, dispersion method, physical adsorption method and ion binding method. This method is characterized by simple operation, but non-covalent The force used by the valence method is relatively weak, and the immobilized enzyme is easy to fall off under high concentration and high ionic strength in industrial production

2) The embedding method can be divided into microcapsule method and grid method. The characteristic of this method is to use sol-gel method, porous fiber or microcapsule to bind the enzyme on the carrier, and make it through physical or chemical force. Stable, usually requires adding enzymes while synthesizing the polymer network structure, the disadvantage of this method is that the material requirements are relatively special, and the universality is not strong

3) Chemical bonding method can be divided into cross-linking method and covalent bonding method. These methods can effectively overcome the shortcomings of non-covalent bonding methods and improve the operational stability of enzymes. The disadvantage is that the use of covalent methods to immobilize enzymes requires After several steps of reaction, for example, first modify the surface of the immobilized material to bring functional groups that can react with the enzyme, and then use the chemical reaction between the enzyme and the functional group to immobilize the enzyme on the surface of the material. For different materials, Due to the different surface properties, different functional groups need to be used for modification, so the steps are cumbersome, and the catalytic activity of the enzyme will be reduced during the immobilization process

[0005] It can be seen from the above that various methods of immobilizing enzymes have their own limitations, especially with the development of bioengineering, it is required to immobilize enzymes on the surface of various materials such as polymer membranes to prepare devices with good performance. The surface of the material has a certain degree of hydrophobicity, and it is difficult for ordinary biomolecules to adsorb to its surface. Although there are some methods such as modifying the substrate surface and then immobilizing enzymes (Chinese patent, publication number CN86100565), this method is only suitable for some The material is suitable, and the enzyme and the material are connected together by using an intermediary polymer (Chinese patent, publication number CN1358856). This method requires the surface of the material to be negatively charged, and to be adsorbed in a solution with a pH higher than the isoelectric point of the enzyme Enzymes have relatively large limitations

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