Process of chitosan-arginine resin anion immobilizing chymotrypsin

Abstract

The invention discloses a method for immobilizing chymotrypsin by chitosan-arginine anionic resin, relating to a method for immobilizing enzyme by a chitosan derivative. The method is to make cross-linked chitosan resin with commercial chitosan as raw material first, then to make chitosan-arginine anionic resin, and finally to perform Schiff reaction to immobilize chymotrypsin to obtain finished products. The method has the advantages of simpleness, easy operation, moderate reaction condition, low production costs, high stability of immobilized chymotrypsin, high recovery of enzyme activity up to 89.95 percent, long half value period which is 18 hours at 75 DEG C and 56 days at 4 DEG C, with optimal pH of 5.92. Products made by the method can be widely used in enzymic reaction in aqueous or nonaqueous medium in medicine, food, light manufacturing and other industries.

Description

technical field [0001] The invention belongs to the technical field of enzyme immobilization, in particular to a method for immobilizing enzymes with chitosan derivatives. technical background [0002] Studies have shown that: chymotrypsin has the functions of digesting pus and necrotic tissue, eliminating wounds, local anti-inflammation, promoting hematoma absorption, and wound healing in medicine; it has the functions of decomposing protein and depilating leather in food and light industries. At present, the free enzymes used mainly have the disadvantages of high cost, low utilization rate, and the discharged waste enzymes pollute the environment. Therefore, the development and use of a new type of immobilized chymotrypsin with a longer half-life has important economic and social value. [0003] Chitosan is a natural polymer material that is abundant in nature and cheap and easy to obtain. Because of its good film-forming, moisturizing, adsorption, and antibacterial prope...

AI Technical Summary

Problems solved by technology

The embedding method refers to embedding enzymes in polymer gel fine grids or semi-permeable membranes. The preparation process is simple and the reaction conditions are relatively mild, and high enzyme activity recovery can be obtained, because only small molecules can pass through polymers. Gel grid, and diffusion resistance will lead to reduced enzyme activity, so the embedding method is only suitable for enzymes that act on small molecule substrates and products

The adsorption method refers to the immobilization method in which the enzyme is adsorbed on an insoluble carrier. This method is easy to operate, with mild conditions, little loss of enzyme activity, and the carrier is cheap and easy to obtain, and can be used repeatedly. However, the interaction between the enzyme and the carrier is weak. , easy to fall off

The covalent binding method refers to the immobilization method in which the enzyme and the carrier are covalently bonded. The immobilized enzyme is not easy to fall off and can be used continuously for a long time, but the activation of the carrier is more complicated, and the binding method is more intense. The reaction caused a large loss of enzyme activity

The cross-linking method refers to the immobilization method of cross-linking between the enzyme and the carrier with a bifunctional or multifunctional reagent. The cross-linking method has good stability of the immobilized enzyme, such as "Chitosan" published in the first issue of the Journal of Ningbo University in March 1999. Preparation of immobilized trypsin and research on its physical and chemical properties” discloses the method of immobilizing trypsin on the chitosan obtained from chitosan crude product under the action of cross-linking agent glutaraldehyde The disadvantage of this method is that it is difficult to obtain pure chitosan from the crude product of chitosan, and chitosan is directly used as a carrier. When the macromolecule of trypsin is close to it, the steric hindrance is large, and the efficiency of direct crosslinking and immobilization of the enzyme is low. Self-crosslinking of trypsin molecules

Another example is the "Preparation Method of Nanomagnetic Chitosan Immobilized Enzyme" patent with publication number CN1904043A, which discloses the use of nanomagnetic chitosan containing acid chloride groups on the surface to react with enzyme protein molecules under the action of a magnetic field to prepare immobilized enzymes. method; the disadvantage of this method is that it is difficult to prepare a nano-magnetic chitosan carrier with a certain particle size, and at the same time, the nano-magnetic chitosan is easily oxidized and precipitated during use, resulting in a reduction in enzyme activity.

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