The immobilized enzyme is a biological reagent with high efficiency, high selectivity and mild reaction conditions.
The immobilized enzyme immobilizes the enzyme onto an inert, low-water-soluble substance, so that the enzyme is immobilized in the reaction, and thus can be easily separated from the reaction system and used again. This technology has great commercial value and has been widely used in industrial enzymatic reactions.
Enzyme immobilization method:
Affinity-tag binding: The enzyme is adsorbed on the outside of the inert substance. In general, this method is the slowest of the three methods listed here. The adsorption reaction is not a chemical reaction, so the active site of the immobilized enzyme may be blocked by the matrix, thereby greatly reducing the enzyme activity.
Entrapment: Enzymes are embedded in insoluble columns or microspheres, such as calcium alginate columns. However, this insolubility hinders substrate entry or product removal.
Cross-linking method: The enzyme is covalently bound to a substrate by a chemical reaction. This method is currently the most effective method. Since the chemical reaction ensures that the binding site does not cover the active site of the enzyme, the activity of the enzyme is only affected by the degree of immobilization.