Immobilized enzyme for magnetic nanometer clay carrier and regenerating method thereof

Abstract

The invention relates to an immobilized enzyme for a magnetic nanometer clay carrier. The immobilized enzyme comprises a carrier with saccharifying enzyme and corresponding immobilized enzyme as immobilizing objects. The immobilized enzyme is prepared by taking a covalence coupling method as an immobilizing method and is characterized in that: the carrier is a magnetic nanometer Fe3O4@ clay compound material; the magnetic nanometer Fe3O4@ clay compound material is composed of magnetic Fe3O4 nanometer grains and a nanometer clay material; and the magnetic Fe3O4 nanometer grains are assembled on the nanometer clay in sequence. A superparamagnetic Fe3O4@ clay compound material is prepared by adopting a solvent heat method according to the invention; the surface of the Fe3O4@ clay compound material is modified by using gamma-aminopropyl triethoxy silane, and then glutaraldehyde is used as a coupling agent for covalently fixing the saccharifying enzyme on the surface of the carrier; two novel carrier regenerating strategies are designed; after the saccharifying enzyme is immobilized by the regenerated carrier, the excellent properties, such as original activity, thermal stability and reusability, of the immobilized enzyme are still maintained; and the immobilized enzyme is also suitable for the regeneration of the immobilized enzyme carrier according to the covalence coupling method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a magnetic nano clay carrier immobilized enzyme and a regeneration method thereof. Background technique [0002] Immobilized enzymes allow repeated or continuous use of enzymes, easy recovery of substrates and products, easy purification, and improve the performance of biocatalysts to a certain extent. In recent years, nanomaterials such as nanoclays have attracted great attention as carriers for immobilized enzymes because of their low cost, small particle size, and unique intercalation properties; another important reason is the silanol on the surface of clays. After activation by different functional groups, it can be used as an attachment site for biomacromolecules. However, the biggest problem in these studies is that the carrier is difficult to separate after the immobilized enzyme completes the catalytic reaction, and the heterogeneous catalysis of magnetic materials can sati...

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Problems solved by technology

However, the biggest problem in these studies is that the carrier is difficult to separate after the immobilized enzyme completes the catalytic reaction, and the heterogeneous catalysis of magnetic materials can satisfy the high-efficiency separation of biocatalysts and the continuous progress of the catalytic process. Therefore, magnetic nanomaterials Modified clay is expected to be an excellent carrier material for immobilized enzymes

[0003] Magnetic iron oxide / clay nanocomposites have been reported in the literature. However, these previous studies have shown that magnetic nanoparticles are often immobilized on the clay surface, and the silanols, which serve as attachment sites for immobilized enzymes, may be covered. This limits the application of magnetic nanoclay materials in the field of immobilized enzymes

In addition, the magnetic strength of these prepared magnetic nanomaterials is also very low and difficult to separate, which greatly limits their applications.

[0004] As far as enzyme immobilization is concerned, in the four main carrier-immobilized enzyme methods of adsorption, cross-linking, embedding and covalent coupling, the immobilized enzyme prepared by the other three methods except the adsorption method cannot be immobilized by the carrier. Regenerated, discarded with the inactivation of the enzyme, which causes environmental pollution, waste of resources and increased production costs

Although the carrier of the immobilized enzyme prepared by the traditional adsorption method can be regenerated, the combination of the carrier and the enzyme is not firm, and the loss of the enzyme is serious, which limits its application.

Therefore, the design, development and preparation of renewable carrier materials has become one of the research priorities of this topic, and there are few reports on the work in this area

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