Method to bind enzyme to carrier using cationic copolymers and product produced thereby
Disclosed are a method of adhering active enzymes to an inert support, the product produced thereby, and a method of using the enzyme-coated support in enzyme-catalyzed reactions such as the glucose isomerase-catalyzed conversion of glucose to fructose. The method includes the steps of coating an inert support with a cationic copolymer, preferably a polyamine, and most preferably a di-C 1 -C 6 -alkylamino-epichlorohydrin copolymer, and then adhering enzyme to the coated support in the absence of any intervening cross-linking agent.
1. A method of making an enzyme adhered on a support, the method comprising:
(a) contacting an inert support with a cationic polymer, whereby the cationic polymer is adhered to the inert support, wherein the cationic polymer comprises a copolymer comprising a di-C 1 -C 6 -alkyl amine co-polymerized with an α-halo-ω-epoxy-C 3 -C 6 -alkane; and then
(b) contacting the inert support from step (a) with an enzyme under conditions wherein the enzyme adheres to the cationic polymer absent any intervening cross-linking agent.
2. The method of claim 1 , wherein in step (a) the inert support comprises a material selected from the group consisting of silica, alumina, titania, diatomaceous earth, kaolin, glass, organic polymers, and cellulose.
3. The method of claim 1 , wherein in step (a) the inert support comprises silica.
4. The method of claim 1 , wherein in step (a) the inert support is a particulate material having an average diameter equal to or smaller than about 30 mesh.
5. The method of claim 1 , wherein in step (a) the inert support is a particulate material having an average diameter equal to or smaller than about 50 mesh.
6. The method of claim 1 , wherein in step (a) the cationic polymer comprises a polyamine.
7. The method of claim 1 , wherein in step (b) the enzyme comprises glucose isomerase.
8. The method of claim 1 , wherein prior to step (a), the inert support is treated with an acid or a chelating agent.
9. The method of claim 8 , wherein prior to step (a), the inert support is treated with an acid, wherein the acid is a citric acid, or treated with a chelating agent, wherein the chelating agent is ethylenediaminetetraacetic acid (EDTA).