A kind of creatine kinase immobilization method

Abstract

The invention provides an immobilized method of creatine kinase. The immobilized method uses chitosan as a raw material, a crosslinking chitosan solution is prepared firstly, a crosslinking chitosan microsphere is prepared, stem grafting lysine crosslinking chitosan resin is prepared then, and finally the stem grafting lysine crosslinking chitosan resin serves as a carrier to immobilize the creatine kinase. The immobilized method has the advantages that conventional production equipment can be used for producing phosphocreatine under a soft condition, a process is simple, operation is simple and convenient, cost of production is low, work efficiency is improved, and a productive process is favorable for environment protection. Stability of prepared immobilized creatine kinase on conditions such as heat, acid, alkali and an organic solvent is obviously improved.

Description

technical field [0001] The invention relates to the technical field of creatine kinase immobilization preparation, in particular to a method for immobilizing creatine kinase with grafted lysine cross-linked chitosan microspheres. Background technique [0002] Creatine kinase is an important kinase that is directly related to intracellular energy operation, muscle contraction, and ATP regeneration. It can reversibly catalyze the transphosphoryl reaction between creatine and ATP. It is used for the early diagnosis of myocardial infarction, the determination of various muscle diseases, and the preparation of creatine phosphate, etc. Free creatine kinase is prepared from rabbit muscle, which is expensive, unstable in solution, easily denatured and inactivated, difficult to separate and recover after reaction, and cannot be reused. This greatly limits the application of creatine kinase, especially in the reaction of preparing creatine phosphate using creatine kinase as a catalys...

AI Technical Summary

Problems solved by technology

This method uses maleic anhydride to cross-link chitosan amino groups, and urease is immobilized in the microspheres while the maleylated chitosan microsphere immobilization carrier is prepared. At the same time as the free amino group of sugar reacts, it also reacts with the free amino group of urease, and a cross-linking reaction between the free amino groups of urease also occurs. A part of the immobilized urease is directly cross-linked on the succinyl skeleton. Disturb the conformation of immobilized urease, significantly reduce the affinity between immobilized urease and substrate; part of the immobilized urease produced by maleic anhydride self-crosslinking, due to the significant increase in volume, resulting in a significant increase in its "rigidity" and a significant decrease in catalytic activity ; In addition, part of the free amino groups of chitosan and maleic anhydride are cross-linked to form a cross-linked chitosan carrier, which significantly reduces the free amino groups of chitosan involved in the immobilization of urease, resulting in a small immobilization load; in addition, the maleic anhydride carbon chain Shorter and harsher reaction conditions with free amino groups, therefore, the urease protein prepared with maleic anhydride as a cross-linking agent has a low coupling rate, low activity recovery, low affinity with the substrate, and low catalytic activity