Method for immobilizing enzymes

• US 4,226,938 A
• Filed: 09/21/1978
• Issued: 10/07/1980
• Est. Priority Date: 07/23/1975
• Status: Expired due to Term

• Current Assignee: Japan Atomic Energy Research Institute (Japan Atomic Energy Agency)
• Original Assignee: Japan Atomic Energy Research Institute (Japan Atomic Energy Agency)
• Inventors: Yoshida, Masaru, Kaetsu, Isao, Kumakura, Minoru
• Primary Examiner(s): Shapiro, Lionel M.
• Application Number:
  US05/944,444
• Time in Patent Office: 747 Days
• Field of Search: 195/63, 195/68, 195/DIG. 11, 435/176, 435/182
• US Class Current: 435/176
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres

Abstract

Enzymes are adsorbed on an adsorbent powder in an aqueous medium and a water-miscible monomer is added thereto, and said monomer is polymerized to form a porous gel lump through which a substrate solution can freely pass. The thus formed enzyme composition has long-lasting activity and thus the enzymes can be easily separated from the reaction system and can be used repeatedly.

Claims

1. A method for immobilizing enzymes or enzyme-containing cells comprising (a) adsorbing said enzymes or enzyme-containing cells on an adsorbent selected from the group consisting of inorganic natural earth adsorbents, synthetic inorganic adsorbents and active carbon in an aqueous medium, (b) admixing the aqueous dispersion of said adsorbent on which enzymes or enzyme-containing cells have been adsorbed with a polymerizable monomer selected from the groups consisting of compounds represented by the general formula ##STR3## wherein R¹ is a group --CH₂ CH₂ -- or --CH(CH₃)CH₂ --, R² is hydrogen or methyl or a group represented by ##STR4## X is hydrogen or methyl, n is an integer 2, 3 or 4 and m is an integer from 2 to 20 and (c) effecting polymerization at a temperature between -20°

C. and -80°

C., to form a porous gel lump in which enzymes or enzyme-containing cells are distributed adsorbed on inorganic adsorbent, and through which a substrate solution can freely pass.

2. The method as defined in claim 1, wherein said adsorbent is selected from the group consisting of active terra alba, bentonite, kaolin, alumina, silica gel, molecular sieves and active carbon, and said monomer B is selected from the compound represented by the general formula:

##STR5## wherein R₁ is --CH₂ CH₂ --, R² is hydrogen, X is hydrogen or methyl, n is 2, 3 or 4, and m is 2, 3 or 4.

3. The method as defined in claim 2, wherein said adsorbent is selected from the group consisting of silica gel, kaolin and active carbon.

4. The method as defined in claim 3, wherein said B monomer is selected from the group consisting of 2-hydroxyethyl methacrylate and triethyleneglycol monomethacrylate.

5. The as defined in claim 1, wherein said B monomer contains a second monomer C having water-miscibility different from that of said Group B monomer in amount up to 30% of the total weight of the monomers.

6. The method as defined in claim 5, wherein said adsorbent is selected from the group consisting of active terra alba, bentonite, kaolin, alumina, silica gel, molecular sieves, and active carbon and said water-miscible monomer is selected from the compounds represented by the general formula:

##STR6## wherein R¹ is --CH₂ CH₂ --, R² is hydrogen, X is hydrogen or methyl, n is 2, 3 or 4, and m is 2, 3 or 4, and said second monomer is a polyfunctional partly water-miscible monomer.

7. The method as defined in claim 6, wherein said adsorbent is selected from the group consisting of silica gel, kaolin, and active carbon.

8. The method as defined in claim 7, wherein said monomer B is selected from the group consisting of 2-hydroxyethyl methacrylate and triethyleneglycol monomethacrylate, and the second monomer is selected from the group consisting of methylolacrylamide, diallyl itaconate, ethyleneglycol dimethacrylate, hexanediol diacrylate, vinylpyrrolidone, trimethylolpropane trimethacrylate.

9. A porous three dimensional reticular structure comprising adsorbed enzyme or enzyme-containing cells whenever obtained by (a) adsorbing said enzymes or enzyme-containing cells on an adsorbent selected from the group consisting of inorganic natural earth adsorbents, synthetic inorganic adsorbents and active carbon in an aqueous medium, (b) admixing the aqueous dispersion of said adsorbent on which enzymes or enzyme-containing cells have been adsorbed with a polymerizable monomer selected from the groups consisting of compounds represented by the general formula ##STR7## wherein R¹ is a group --CH₂ CH₂ -- or --CH(CH₃)CH₂ --, R² is hydrogen or methyl or a group represented by ##STR8## X is hydrogen or methyl, n is an integer 2, 3 or 4 and m is an integer from 2 to 20 and (c) effecting polymerization at a temperature between -20°

C. and -80°

C., to form a porous gel lump in which enzymes or enzyme-containing cells are distributed adsorbed on inorganic adsorbent, and through which a substrate solution can freely pass.