# US 10,246,786 B2 · Renewable bioelectronic interface for electrobiocatalytic reactor

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- Number: US 10,246,786 B2
- Document ID: US10246786B2
- Status: Granted
- Kind: B2
- Application number: 15/177,668
- Filed: 2016-06-09
- Published: 2016-11-10
- Granted: 2019-04-02
- Earliest claimed: 2009-04-24
- Assignees: Board of Trustees of Michigan State University
- Applicants: Board of Trustees of Michigan State University
- Inventors: Robert Mark Worden; Brian L. Hassler; Lawrence T. Drzal; IIsoon Lee
- Web page: https://patentgrep.com/patent/US10246786B2

## CPC

- C25B 11/095: at least one of the compounds being organic
- C12Q 1/005: {involving specific analytes or enzymes (including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence)}
- C25B 11/051: Electrodes formed of electrocatalysts on a substrate or carrier
- C25B 11/057: consisting of a single element or compound
- G01N 27/3272: {Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels (optical biosensors G01N33/52)}
- H01M 4/9008: {Organic or organo-metallic compounds}
- H01M 8/16: Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
- Y02E 60/50: Fuel cells
- C25B: ELECTROLYTIC OR ELECTROPHORETIC PROCESSES FOR THE PRODUCTION OF COMPOUNDS OR NON-METALS; APPARATUS THEREFOR (anodic or cathodic protection C23F13/00; single-crystal growth C30B)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- H01M: PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## 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

### Claim 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.

### Claim 2 (depends on 1)

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

### Claim 3 (depends on 2)

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

### Claim 4 (depends on 1)

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

### Claim 5 (depends on 1)

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.

### Claim 6 (depends on 1)

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

### Claim 7 (depends on 6)

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

### Claim 8 (depends on 7)

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

### Claim 9 (depends on 1)

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

### Claim 10 (depends on 9)

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.

### Claim 11 (depends on 10)

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

### Claim 12 (depends on 9)

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.

### Claim 13 (depends on 9)

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.

### Claim 14 (depends on 12)

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

### Claim 15 (depends on 1)

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

### Claim 16 (depends on 1)

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.

### Claim 17 (depends on 16)

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

### Claim 18 (depends on 16)

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.

### Claim 19 (depends on 16)

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

### Claim 20 (depends on 1)

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.

### Claim 21 (depends on 20)

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

### Claim 22 (depends on 21)

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

### Claim 23 (depends on 22)

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

### Claim 24 (depends on 22)

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.

### Claim 25 (depends on 1)

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

### Claim 26 (depends on 1)

The device of claim 1 containing no thiol linkages.

### Claim 27 (depends on 1)

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

### Claim 28 (depends on 2)

The device of claim 2, wherein the polyelectrolyte provides enzymatic surface coverage up to about 3.3×10−11 moles/cm2.

### Claim 29 (depends on 16)

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

### Claim 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.