Renewable bioelectronic interface for electrobiocatalytic reactor

• US 10,246,786 B2
• Filed: 06/09/2016
• Issued: 04/02/2019
• 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, IIsoon
• Primary Examiner(s): Hobbs, Michael L
• Application Number:
  US15/177,668
• Publication Number:
  US 20160326658A1
• Time in Patent Office: 1,027 Days
• Field of Search: 204400, 204252, 20429011, 20429007, 977742, 977734, 427458, 4352879
• 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 carbon electrode; and

a bioelectronic interface bonded to a surface of the electrically conductive carbon 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 via 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 polyelectroly.

3. The device of claim 2, wherein the enzyme is an oxidoreductase, a dehydrogenase, a secondary alcohol dehydrogenase or a mannitol dehydrogenase.

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

5. 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 carbon 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 carbon electrode.

6. The device of claim 1 wherein the electrically conductive carbon electrode further includes a carbon substrate.

7. The device of claim 6, wherein the carbon substrate is a vitreous carbon substrate.

8. The device of claim 7, wherein the vitreous carbon substrate is a reticulated vitreous carbon electrode.

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

10. The device of claim 9, wherein the bioelectronic interface includes an electron mediator that reduces the electrical potential needed to transfer electrons during a chemical reaction.

11. The device of claim 10, wherein the electron mediator is toluidine blue O, neutral red or Nile blue A.

12. The device of claim 9, wherein the redox cofactor is covalently bound to a polyelectrolyte that is electrostatically bound directly or indirectly to the surface of the electrically conductive carbon electrode.

13. The device of claim 9, 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.

14. The device of claim 12, wherein a boronate linkage is used to bind the redox cofactor to said polyelectrolyte.

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

16. 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.

17. The device of claim 16, wherein a redox cofactor is associated with PEMs that are bound together by alternating layers of oppositely charged polyelectrolytes.

18. The device of claim 16, wherein a redox cofactor and an electron mediator are associated with PEMs that are bound together by alternating layers of oppositely charged polyelectrolytes.

19. The device of claim 16, wherein the oppositely charged polyelectrolytes are polyethyleneimine (PEl) and polyacrylic acid (PAA).

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

21. The device of claim 20, wherein the catalytically active material includes at least two different catalytically active components.

22. The device of claim 21, wherein said catalytically active components comprise different enzymes that catalyze different reactions.

23. The device of claim 22 wherein said enzymes comprise a xylose isomerase and a mannitol dehydrogenase.

24. The device of claim 22, 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 cofactors.

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

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

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

28. The device of claim 2, wherein the polyelectrolyte provides enzymatic surface coverage up to about 3.3×

10<sup>−

11</sup> moles/cm².

29. The device of claim 16 wherein each of the PEMs has a thickness between about one and about five nanometers.

30. A method comprising:

producing a bioelectronic device comprising bonding a bioelectronic interface to a surface of an electrically conductive carbon electrode, wherein the bioelectronics interface is 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 via 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.