Composite of enzyme and carbon structure complex, method for producing the same and use thereof

• US 9,074,174 B2
• Filed: 05/11/2011
• Issued: 07/07/2015
• Est. Priority Date: 05/20/2010
• Status: Active Grant

• Current Assignee: Korea University Research and Business Foundation (Korea University)
• Original Assignee: Korea University Research and Business Foundation (Korea University)
• Inventors: Kim, Jungbae
• Primary Examiner(s): Ryan, Patrick
• Assistant Examiner(s): COLUCCI RIOS, JOSE A
• Application Number:
  US13/698,743
• Publication Number:
  US 20130143130A1
• Time in Patent Office: 1,518 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12N 11/02 Enzymes or microbial cells immobilised on or in an organic carrier
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: H01M 8/16 Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
• CPC Class Codes: Y02E 60/50 Fuel cells
• CPC Class Codes: Y02P 70/50 Manufacturing or production processes characterised by the final manufactured product

Abstract

Disclosed is a composite of enzyme and carbon structure. In the composite of enzyme and carbon structure, a significantly large amount of an enzymeis immobilized on the surface of carbon structures without the formation of chemical bonds (particularly, covalent bonds) between the enzyme molecules and the carbon structures. Since the surface of the carbon structures does not need to be modified to form chemical bonds, the electrical conductivity of the composite of enzyme and carbon structure is not reduced and the stability of the composite is maintained high even after the passage of a long time in various environments. Therefore, the use of the composite of enzyme and carbon structure enables the fabrication of various devices, such as biosensors and biofuel cells, with markedly improved performance as compared to the use of conventional enzyme/carbon structure composites.

Claims

1. A method for producing a composite of enzyme and carbon structure, comprising:

(1) dispersing elongated carbon structures in an aqueous enzyme-solution comprising an enzyme so that the enzyme molecules adsorb on a surface of the carbon structures, wherein the surface of the carbon structures is not modified to form functional groups capable of forming covalent bonds with the enzyme molecules;

(2) adding a precipitant to the enzyme solution to cause the enzyme molecules to precipitate and to deposit on the carbon structure; and

(3) adding a cross-linker to cross-link the precipitated enzyme molecules, wherein the cross-linked enzyme molecules surround the carbon structures, and wherein there is no covalent bonding between the carbon structures and the enzyme molecules.

2. The method according to claim 1, wherein the carbon structures have a diameter in a micrometer or nanometer range.

3. The method according to claim 2, wherein the carbon structures are selected from carbon tubes, carbon rods, and carbon wires.

4. The method according to claim 1, wherein electron conductivity of the carbon structures is preserved.

5. The method according to claim 1, wherein the precipitant is selected from aqueous solutions of methanol, ethanol, 1-propanol, 2-propanol, butyl alcohol, acetone, PEG, ammonium sulfate, sodium chloride, sodium sulfate, sodium phosphate, potassium chloride, potassium sulfate and potassium phosphate, and mixtures thereof.

6. The method according to claim 1, wherein the precipitant has a concentration of 10 to 50% by weight.

7. The method according to claim 1, wherein the cross-linker comprises one or more compounds selected from the group consisting of diisocyanates, dianhydrides, diepoxides, dialdehydes, diimides, 1-ethyl-3-dimethylaminopropylcarbodiimide, glutaraldehyde, bis(imido esters), bis(succinimidyl ester), and diacid chlorides.

8. The method according to claim 1, wherein the enzyme molecules form shells surrounding the carbon structures without the formation of covalent bonds between the enzyme and the carbon structures.

9. The method according to claim 1, wherein the enzyme is selected from the group consisting of chymotrypsin, trypsin, subtilisin, papain, lipase, horse radish peroxidase, soybean peroxidase, chloroperoxidase, manganese peroxidase, tyrosinase, laccase, cellulase, xylanase, lactase, sucrase, organophosphohydrolase, cholin esterase, glucose oxidase, alcohol dehydrogenase, glucose dehydrogenase, hydrogenase, glucose isomerase, and mixtures thereof.

10. A composite of enzyme and carbon structure comprising:

elongated carbon structures, wherein a surface of the carbon structures is not modified to introduce functional groups capable of forming covalent bonds with enzyme molecules; and

one or more shells comprising crosslinked enzyme molecules surrounding the carbon structures,

wherein there is no covalent bonding between the enzyme molecules and the carbon structures.

11. The composite according to claim 10, wherein the carbon structures are selected from carbon nanotubes, carbon nanorods, and carbon nanowires.

12. The composite according to claim 10, wherein electron conductivity of the carbon structures is preserved.

13. The composite according to claim 10, wherein each of the one or more shells has a thickness of 10 to 1,000 nm.

14. A device selected from the group consisting of biosensors, biofuel cells, enzyme columns, ELISA kits, bioremediation devices, antifouling agents, and ibuprofen production, wherein the device comprises the composite according to claim 10.