Biomimetic immobilization method of multienzyme system

Abstract

The invention relates to a technique for immobilizing a multienzyme system, which comprises the following steps: 1) preparation of precursor solution, namely, adding TMOS (tetramethyl orthosilicate) into a 1 mmol / L HCl solution at room temperature, adding a phosphate buffer of which the pH value is 5-9 after the TMOS is completely dissolved and the solution becomes clear, and evenly mixing; 2) preparation of immobilized enzyme: evenly mixing the multienzyme solution with a PDADMA (polydimethyldiallyl ammonium chloride) solution to obtain an enzyme-containing solution A; and under the 30 DEG C homothermal condition, adding the pretreated precursor solution into the solution A, immediately shaking, standing at room temperature, carrying out centrifugal separation, washing with the phosphate buffer, and carrying out vacuum freeze drying on the residual solid for 24 hours, thereby obtaining the biomimetic silicified multienzyme. The method provided by the invention has the advantages of simple preparation technique, mild conditions, low loss of enzyme activity, and high enzyme activity recovery rate (up to 85.12%); and the carrier is the TMOS which is cheap and accessible.

Description

Technical field [0001] The invention belongs to the field of immobilized enzymes, in particular to a process method for immobilized multi-enzyme systems. Background technique [0002] Multi-enzyme system refers to a whole structure and function composed of different enzymes, which can carry out continuous reaction catalysis. The material metabolism of organisms in nature is maintained by various multi-enzyme systems. The multi-enzyme system is Efficient and precise systems often consist of two or more enzymes forming a complex. The product catalyzed by the first enzyme in the complex is directly catalyzed by the second enzyme as the substrate, and the product catalyzed by the second enzyme is the substrate of the next enzyme in the complex. The continuous catalytic reaction is carried out in this way to make the catalytic efficiency significant Improve and organically coordinate the balance between a series of enzyme reactions. The high efficiency and high coordination of the m...

AI Technical Summary

Problems solved by technology

[0006] The above related literatures have investigated the immobilized enzymes prepared by different methods in the multi-enzyme system. The immobilization methods used involve adsorption, cross-linking and covalent bonding. The efficiency of the immobilized enzyme is low, and the stability of the immobilized enzyme is weak; covalent bonding and cross-linking method rely on chemical reactions to realize the combination of the carrier and the enzyme during the immobilization process, which has a great impact on the activity of the enzyme. Therefore, an immobilization method is sought A method with high efficiency and low impact on enzyme activity is important

So far, there are no relevant reports in the literature on the co-immobilization of multi-enzyme systems using the biomimetic siliconization embedding method