IMMOBILIZED ORGANOPHOSPHATE-DEGRADING ENZYMES AND METHODS OF MAKING THE SAME

• US 20130316431A1
• Filed: 11/13/2012
• Published: 11/28/2013
• Est. Priority Date: 11/11/2011
• Status: Abandoned Application

• Current Assignee: Augustine A. Dinovo, David A. Schofield, Dominic P. Dinovo, Francis H. Verhoff, Matthew F. Smiechowski
• Original Assignee: Augustine A. Dinovo, David A. Schofield, Dominic P. Dinovo, Francis H. Verhoff, Matthew F. Smiechowski
• Inventors: DiNovo, Augustine A., DiNovo, Dominic P., Schofield, David A., Smiechowski, Matthew F., Verhoff, Francis H.
• Application Number:
  US13/675,576
• Publication Number:
  US 20130316431A1
• Time in Patent Office: Days
• US Class Current: 435/179
• CPC Class Codes: C12N 11/02 Enzymes or microbial cells immobilised on or in an organic carrier
• CPC Class Codes: C12N 11/06 attached to the carrier via a bridging agent
• CPC Class Codes: C12N 11/10 the carrier being a carbohydrate
• CPC Class Codes: C12N 11/12 Cellulose or derivatives thereof

Abstract

The present application discloses immobilized enzymes and immobilized enzyme materials comprising a crosslinked organophosphate-degrading enzyme having a support material which includes a biomass material and/or a polymeric material. The resulting immobilized enzyme materials may be biodegradable. The present application also discloses methods of making and using the disclosed immobilized organophosphate hydrolase enzyme and enzyme materials.

Claims

1. An immobilized enzyme material comprising a crosslinked organophosphate-degrading enzyme having a support matrix that includes a biomass material, wherein the crosslinked organophosphate-degrading enzyme was formed by reacting an organophosphate-degrading enzyme with at least two polyfunctional materials.

2. The immobilized enzyme material of claim 1, wherein at least one of the two polyfunctional materials is selected from the group consisting of a di-aldehyde, disuccinimidyl suberate, an organic di-acid, glutaraldehyde, dimethyl pimelimidate, cyanuric chloride, succinic acid, hexamethylene diisocyanate, diimidoester, triazine, diisocyanate, and di(n-hydroxysuccinimide ester).

3. The immobilized enzyme material of claim 1, wherein at least one of the two polyfunctional materials is a polyamine.

4. The immobilized enzyme material of claim 3, wherein at least one of the two polyfunctional materials which is an polyamine is selected from the group consisting of polyethylenimine, polypyrrole, chitosan, a protein, and gelatin.

5. The immobilized enzyme material of claim 4, wherein the biomass material is selected from the group consisting of bacterial cell material, fungal cell material, cellulose, dextran, starch agar, alginate, carrageenans, collagen, gelatin, albumin, ferritin, cotton, chitin, exoskeleton, and wool.

6. The immobilized enzyme material of claim 1, wherein the immobilized enzyme material is biodegradable.

7. The immobilized enzyme material of claim 1, wherein the biomass material is different from the biomass material the organophosphate-degrading enzyme was derived from.

8. The immobilized enzyme material of claim 1, wherein at least one of the two polyfunctional materials includes an additional biomass material having a plurality of amine groups thereon.

9. The immobilized enzyme material of claim 8, wherein the at least one polyfunctional material which includes a biomass material is selected from the group consisting of bacterial cell material, fungal cell material, cellulose, dextran, starch agar, alginate, carrageenans, collagen, gelatin, albumin, ferritin, cotton, chitin, exoskeleton, and wool.

10. The immobilized enzyme material of claim 1, wherein the biomass material is insoluble.

11. The method of claim 1, wherein the organophosphate-degrading enzyme is selected from the group consisting of aryldialkylphosphatases (E.C. 3.1.8.1) and diisopropyl-fluorophosphatase (E.C. 3.1.8.2).

12. A method for preparing an immobilized organophosphate-degrading enzyme material comprising:

providing a biomass material, an aqueous suspension of an organophosphate-degrading enzyme, and at least two polyfunctional materials;

adding the biomass material, and one of the two polyfunctional materials to the aqueous suspension of the organophosphate-degrading enzyme to form a reaction mixture;

adding the second polyfunctional material to the reaction mixture to flocculate the immobilized organophosphate-degrading enzyme material.

13. The method of claim 12, wherein the method further involves stirring the reaction mixture before adding the second polyfunctional material to the reaction mixture.

14. The method of claim 12, wherein providing a biomass material and the at least two polyfunctional materials involves providing a biodegradable biomass material and biodegradable polyfunctional materials.

15. The method of claim 12, wherein providing a biomass material involves providing a biomass material different from the biomass material that produced the enzyme.

16. The method of claim 12, wherein providing a biomass material involves providing a biomass material is selected from the group consisting of bacterial cell material, fungal cell material, cellulose, dextran, starch agar, alginate, carrageenans, collagen, gelatin, albumin, ferritin, cotton, chitin, exoskeleton, and wool.

17. The method of claim 12, wherein providing at least two polyfunctional materials involves providing an additional biomass material having a plurality of amine groups thereon.

18. The method of claim 12, wherein at least one of the two polyfunctional materials is selected from the group consisting of a di-aldehyde, disuccinimidyl suberate, an organic di-acid, glutaraldehyde, dimethyl pimelimidate, cyanuric chloride, succinic acid, hexamethylene diisocyanate, diimidoester, triazine, diisocyanate, and di(n-hydroxysuccinimide ester).

19. The method of claim 12, wherein providing at least two polyfunctional materials involves providing a polyamine selected from the group consisting of polyethylenimine, polypyrrole, chitosan, a protein, and gelatin.

20. The method of claim 19, wherein the biomass material is selected from the group consisting of bacterial cell material, fungal cell material, cellulose, dextran, starch agar, alginate, carrageenans, collagen, gelatin, albumin, ferritin, cotton, chitin, exoskeleton, and wool.

21. The method of claim 12, wherein the organophosphate-degrading enzyme is selected from the group consisting of aryldialkylphosphatases (E.C. 3.1.8.1) and diisopropyl-fluorophosphatase (E.C. 3.1.8.2).

22. The method of claim 12, wherein providing a biomass material involves providing an insoluble biomass material.

23. The method of claim 12, wherein at least one of the two polyfunctional materials is a polyamine.

24. A method for decontaminating an area containing a material susceptible to enzymatic degradation comprising:

providing a biodegradable immobilized enzyme material in a form suitable for application to an area and adapted to degrade the material;

contacting the area with the form of the biodegradable immobilized enzyme material;

wherein upon degradation of the material, the biodegradable immobilized enzyme material itself biodegrades, making its removal unnecessary.

25. A method for transforming a material susceptible to enzymatic transformation comprising:

providing a biodegradable immobilized enzyme material adapted to transform the material in a form suitable for use in a reaction chamber;

placing the immobilized enzyme material in a reaction chamber;

contacting the material to be transformed with the immobilized enzyme material in the reaction chamber;

removing the immobilized enzyme material from the reaction chamber to an area where the immobilized enzyme material may biodegrade.

26. The method of claim 25, wherein the reaction chamber is a reaction column.

27. The method of claim 25, wherein the reaction chamber is a closed reaction vessel.

28. The method of claim 25, wherein the reaction chamber is a packed bed.