Abstract A general process for preparing water-insoluble enzymes which comprises covalently bonding said enzymes to an insoluble polymer, said bonding taking place by the reaction of imidoester functional groups on said polymer with enzyme amino groups. The polymer is preferably an acrylonitrile-based polymer wherein at least a substantial portion of the surface acrylonitrile groups are converted into imidoesters by contacting with an alcohol and a hydrogen halide (e.g., HCl) to form imidoester functional groups. Said imidoester functional groups are then contacted with an aqueous enzyme-containing solution at reaction conditions whereby an insoluble enzyme composite is formed. Said insoluble enzyme composite retains activity substantially equivalent to the enzyme in its native state and further shows increased resistance to degradation by heat as well as chemical denaturants.
The process of claim 1 wherein said contacting takes place in an aqueous alkaline solution.
Claim 2
Dependent Claim
18 words
The process of claim 2 wherein said contacting takes place at a temperature of from 15*C to 50*C.
Claim 3
Dependent Claim
20 words
The process of claim 2 wherein said enzyme is selected from the group consisting of the hydrolases and the oxidoreductases.
Claim 4
Dependent Claim
11 words
The process of claim 2 wherein said enzyme is Alpha -chymotrypsin.
Claim 5
Dependent Claim
10 words
The process of claim 2 wherein said enzyme is trypsin.
Claim 6
Dependent Claim
10 words
The process of claim 2 wherein said enzyme is phenolase.
Claim 7
Dependent Claim
10 words
The process of claim 2 wherein said enzyme is ribonuclease.
Claim 8
Dependent Claim
15 words
The process of claim 2 wherein said polymer is in the form of a fiber.
Claim 9
Dependent Claim
25 words
The process of claim 1 wherein said alcohol is selected from the group consisting of methanol and ethanol and the hydrogen halide is hydrogen chloride.
Claim 10
Dependent Claim
22 words
The process of claim 10 wherein the acrylonitrile monomer content of said acrylonitrile containing polymer is at least 25 percent by weight.
Claim 11
Dependent Claim
25 words
The process of claim 11 wherein said insoluble polymer is reacted with said alcohol and hydrogen halide at a temperature of from -25*C to +75*C.
Claim 12
Dependent Claim
70 words
An enzyme composite comprising an enzyme covalently coupled to a polymer, said enzyme being coupled with said polymer through an amidine linkage, said amidine linkage being provided through the reaction of an imidate ester functional group present on said polymer with an amino group present on said enzyme, and said imidate ester functional group being provided through the reaction of an acrylonitrile group present on said polymer with an alcohol.
Claim 13
Dependent Claim
11 words
The product of claim 13 wherein said enzyme is Alpha -chymotrypsin.
Claim 14
Dependent Claim
10 words
The product of claim 13 wherein said enzyme is trypsin.
Claim 15
Dependent Claim
10 words
The product of claim 13 wherein said enzyme is phenolase.
Claim 16
Dependent Claim
10 words
The product of claim 13 wherein said enzyme is ribonuclease.
Claim 17
Dependent Claim
20 words
The product of claim 13 wherein said enzyme is selected from the group consisting of the hydrolases and the oxidoreductases.