Process for preparing immobilized enzymes

A process for increasing the activity and stability of an immobilized enzyme which comprises the steps of:

• (a) treating an inorganic porous support material which possesses pore diameters of from about 100 to about 55,000 Angstroms and a surface area of from about 1 to about 500 m.sup.2 /g with a solution of a first water soluble bifunctional monomer;
• (b) removing unadsorbed monomer;
• (c) contacting the treated support material with a solution containing an excess of a second bifunctional monomeric material of from about 2 to about 50 moles of said bifunctional monomer per mole of said first water soluble bifunctional monomer wherein said second bifunctional monomeric material reacts with said first bifunctional monomer adsorbed on said support material to form a polymeric material and to provide pendent terminally functionalized groups for convalent bonding thereto of an enzyme;
• (d) removing unreacted bifunctional monomeric material;
• (e) treating the resultant inorganic-organic support matrix with an enzyme to covalently bind said enzyme to said support matrix;
• (f) treating the resultant immobilized enzyme-support matrix complex with an additional amount of said second bifunctional monomeric material to provide pendent terminally functionalized groups for covalent bonding thereto of additional enzymes;
• (g) removing excess bifunctional monomeric material;
• (h) adding a substrate for said enzyme to the complex;
• (i) treating said complex with an additional amount of said enzyme at a temperature in the range of from about 0.degree. to about 20.degree. C.;
• (j) removing excess substrate and enzyme;
• and,
• (k) treating said complex with a mixture of said substrate and bifunctional monomeric material.