Electrobiocatalytic reactors having reversible bioelectronic interfaces and methods and devices related thereto

• US 9,406,959 B2
• Filed: 04/23/2010
• Issued: 08/02/2016
• Est. Priority Date: 05/10/2005
• Status: Active Grant

• Current Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Original Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Inventors: Worden, Robert Mark, Hassler, Brian L, Drzal, Lawrence T., Lee, Ilsoon
• Primary Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US12/766,169
• Publication Number:
  US 20100314248A1
• Time in Patent Office: 2,293 Days
• Field of Search: 435/287.9, 435/458, 204/252, 204/400, 204/290.07, 204/290.11, 977/742, 427/458
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence
• CPC Class Codes: C25B 11/051 Electrodes formed of electrocatalysts on a substrate or carrier
• CPC Class Codes: C25B 11/057 consisting of a single element or compound
• CPC Class Codes: C25B 11/095 at least one of the compounds being organic
• CPC Class Codes: G01N 27/3272 Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels optical biosensors G01N33/52
• CPC Class Codes: H01M 4/9008 Organic or organo-metallic compounds
• 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

Abstract

An inexpensive, easily renewable bioelectronic device useful for bioreactors, biosensors, and biofuel cells includes an electrically conductive carbon electrode and a bioelectronic interface bonded to a surface of the electrically conductive carbon electrode, wherein the bioelectronic interface includes catalytically active material that is electrostatically bound directly or indirectly to the electrically conductive carbon electrode to facilitate easy removal upon a change in pH, thereby allowing easy regeneration of the bioelectronic interface.

Claims

1. A bioelectronic device comprising:

an electrically conductive electrode having a carbon substrate; and

a bioelectronic interface bonded to a surface of the electrically conductive electrode and configured to regenerate, the bioelectronic interface including a catalytically active material that facilitates electron transfer, the catalytically active material configured to be releasably and electrostatically bound directly or indirectly to the surface, wherein said interface also contains one or more organic nanoparticles selected from one or more exfoliated nanographite supports, one or more carbon nanotubes, one or more fullerenes and combinations thereof, wherein said organic nanoparticles are incorporated into a polyelectrolyte, wherein the polyelectrolyte is configured to be removably and electrostatically bound to a non-thiol ionic linker that is covalently bound to a carbon atom at the surface.

2. The device of claim 1, wherein the catalytically active material includes an enzyme that is bound, directly or indirectly, to the polyelectrolyte.

3. The device of claim 2, wherein the enzyme is an oxidoreductase.

4. The device of claim 2, wherein the enzyme is a dehydrogenase.

5. The device of claim 2, wherein the enzyme is a secondary alcohol dehydrogenase.

6. The device of claim 2, wherein the enzyme is a mannitol dehydrogenase.

7. The device of claim 1, wherein the covalent bond with a carbon atom is a carbon-nitrogen bond.

8. The device of claim 1, wherein the non-thiol ionic linker is glycine.

9. The device of claim 1, wherein said polyelectrolyte is comprised of a compound that includes a linking moiety covalently bound to the carbon atom at the surface of the electrically conductive electrode, and further includes at least one ionized moiety for achieving the direct or indirect electrostatic bonding of the catalytically active material to the electrically conductive electrode.

10. The device of claim 1, wherein the carbon substrate is a vitreous carbon substrate.

11. The device of claim 10, wherein the vitreous carbon substrate is a reticulated vitreous carbon substrate.

12. The device of claim 1, wherein the bioelectronic interface further comprises a redox cofactor that facilitates or enhances activity of the catalytically active material.

13. The device of claim 12, wherein the bioelectronic interface includes an electron mediator that reduces the electrical potential needed to transfer electrons during a chemical reaction.

14. The device of claim 13, wherein the electron mediator is toluidine blue O.

15. The device of claim 13, wherein the electron mediator is neutral red.

16. The device of claim 13, wherein the electron mediator is Nile blue A.

17. The device of claim 12, wherein the redox cofactor is covalently bound to a polyelectrolyte that is electrostatically bound directly or indirectly to the surface of the electrically conductive electrode.

18. The device of claim 12, wherein the redox cofactor is selected from nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NADP), flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN), and combinations thereof.

19. The device of claim 17, wherein a boronate linkage is used to bind the redox cofactor to said polyelectrolyte.

20. The device of claim 1, wherein the polyelectrolyte is polyethyleneimine.

21. The device of claim 1, wherein the exfoliated nanographite supports comprise exfoliated graphite nanoplatelets.

22. The device of claim 1, wherein the catalytically active material is associated with polyelectrolyte multilayers (PEMs) that are bound together by alternating layers of oppositely charged polyelectrolytes.

23. The device of claim 22, wherein a redox cofactor is associated with PEMs that are bound together by alternating layers of oppositely charged polyelectrolytes.

24. The device of claim 22, wherein a redox cofactor and an electron mediator are associated with PEMs that are bound together by alternating layers of oppositely charged polyelectrolytes.

25. The device of claim 22, wherein the oppositely charged polyelectrolytes are polyethyleneimine (PE1) and polyacrylic acid (PAA).

26. The device of claim 1, wherein the polyelectrolyte ionically bonds a dehydrogenase enzyme and a redox cofactor to an ionically functionalized, electrically conductive electrode to which an electron mediator is bound.

27. The device of claim 26, wherein the catalytically active material includes at least two different catalytically active components.

28. The device of claim 27, wherein said catalytically active components comprise different enzymes that catalyze different reactions.

29. The device of claim 28, wherein said enzymes comprise a xylose isomerase and a mannitol dehydrogenase.

30. The device of claim 28, wherein a branched linking moiety and the polyelectrolyte are used to couple the enzymes, and optionally couple a redox cofactor for one or both of the enzymes, and/or optionally couple an electron mediator for one or both of the redox co factors.

31. The device of claim 1, wherein said organic nanoparticles provide enzymatic surface coverage up to about 3.3×

10<sup>−

11</sup> moles/cm².

32. The device of claim 1 containing no thiol linkages.

33. The device of claim 1, wherein said interface is molecularly self-assembled.

34. The device of claim 22, wherein each of the PEMs has a thickness between about one and about five nanometers.

43. The device of claim 1 further comprising an electrobiocatalytic reactor.

35. A bioelectronic device comprising:

an electrically conductive electrode having a carbon substrate; and

a bioelectronic interface configured to regenerate and containing at least one polyelectrolyte layer bonded to a surface of the electrically conductive electrode, the bioelectronic interface including a catalytically active material that facilitates electron transfer, the catalytically active material electrostatically bound directly or indirectly to the electrically conductive electrode, wherein said interface also contains one or more organic nanoparticles selected from one or more exfoliated nanographite supports, one or more carbon nanotubes, one or more fullerenes and combinations thereof, wherein said organic nanoparticles are incorporated into a polyelectrolyte which is configured to be removably and electrostatically bound to a non-thiol ionic linker that is covalently bound to a carbon atom at the surface.

36. The device of claim 35 wherein each of said polyelectrolyte layers has a thickness from about one and about five nanometers.

37. The device of claim 35 containing no thiol linkages.

38. A method comprising:

providing an electrically conductive carbon electrode having a carbon substrate; and

bonding a bioelectronic interface bonded to a surface of the electrically conductive electrode and configured to regenerate, the bioelectronic interface including a catalytically active material that facilitates electron transfer, the catalytically active material configured to be releasably and electrostatically bound directly or indirectly to the surface, wherein said interface also contains one or more organic nanoparticles selected from one or more exfoliated nanographite supports, one or more carbon nanotubes, one or more fullerenes and combinations thereof, wherein said organic nanoparticles are incorporated into a polyelectrolyte which is configured to be removably and electrostatically bound to non-thiol ionic linker that is covalently bound to a carbon atom at the surface.

39. The method of claim 38 wherein the exfoliated nanographite supports comprise exfoliated graphite nanoplatelets.

40. The method of claim 38 further comprising releasing the catalytically active material from the surface via exposure of said interface to a first pH change.

41. The method of claim 40 further comprising regenerating additional catalytically active material via exposure of said interface to a second pH that facilities electrostatic bonding of the catalytically active material to the surface.

42. The method of claim 38 wherein the bioelectronics device is contained within a reactor and the regeneration occurs without removing said electrode from the reactor by flowing said aqueous mediums through the reactor.

44. A bioelectronic device comprising:

an electrically conductive electrode having a carbon substrate; and

a bioelectronic interface bonded to a surface of the electrically conductive electrode and configured to regenerate, the bioelectronic interface including a catalytically active material that is associated with polyelectrolyte multilayers (PEMs) that are bound together by alternating layers of oppositely charged polyelectrolytes comprising polyethyleneimine (PE1) and polyacrylic acid (PAA) and that facilitates electron transfer, the catalytically active material configured to be releasably and electrostatically bound directly or indirectly to the surface, wherein said interface also contains one or more organic nanoparticles selected from one or more exfoliated nanographite supports, one or more carbon nanotubes, one or more fullerenes and combinations thereof, wherein said organic nanoparticles are incorporated into a polyelectrolyte which is configured to be removably and electrostatically bound to an ionic linker that is covalently bound to a carbon atom at the surface with a carbon-nitrogen bond.

45. An electrobiocatalytic reactor comprising:

a cathode compartment containing a first electrolytic solution and an anode compartment containing a second electrolytic solution, the anode compartment and cathode compartments separated by a proton permeable membrane;

a cathode located in the cathode compartment and in contact with the first electrolyte, and an anode located in the anode compartment and in contact with the second electrolyte, wherein at least one of the anode and cathode being an electrically conductive electrode; and

a bioelectronic interface bonded to a surface of the electrically conductive electrode, the bioelectronic interface including a catalytically active material that facilitates electron transfer, the catalytically active material configured to be releasably and electrostatically bound directly or indirectly to the surface, wherein said interface also contains one or more organic nanoparticles selected from one or more exfoliated nanographite supports, one or more carbon nanotubes, one or more fullerenes and combinations thereof, wherein said organic nanoparticles are incorporated into a polyelectrolyte which is configured to be removably and electrostatically bound to a non-thiol ionic linker that is covalently bound to a carbon atom at the surface.

46. The reactor of claim 45, wherein both the cathode and the anode are electrically conductive electrodes, each having a carbon substrate, wherein the cathode has a surface with a bioelectronic interface comprising xylose isomerase and mannitol dehydrogenase bound thereto and the anode has a surface with a bioelectronic interface comprising glycerol dehydrogenase bound thereto, wherein the reactor is configured to simultaneously produce mannitol from glucose and dihydroxyacetone from glycerol.