Download BE REU @ SLU Department of Chemistry Dr. Shelley D. Minteer

Enzymatic Glucose Biofuel Cell:
Concentration Studies
and Biocompatibility
Joe Fazio
BE REU @ SLU
Dr. Shelley D. Minteer
Kyle Sjöholm
SLU Department of Chemistry
Background
Enzymatic Biofuel cell:




Enzymes
Power biomedical
devices
High power and current
density
Incomplete oxidation
www.nano-biokit.com
Biofuel Cell Process




Reaction at anode produces
protons
Electrons create current
Protons diffuse to cathode
Protons at cathode react
with oxygen
Mediated Electron Transfer (MET)
Electrocatalyst

Commonly used to
reduce overpotential
2e-
Glucose
NADH

Facilitates ion
transfer to electrode
Gluconolactone
NAD+
Glucose
Dehydrogenase
Entrapped in
Polymer
060 Toray
Paper
Electrode
Modified Polymers


Immobilize enzymes
Extend functional lifetime
Microencapsulation:
 Support enzyme structure
Neutral pH
 Micellar environment
 Geometry
 Ion exchange
properties

Polymer encapsulation
Project Goals

Power Densities



Hypoglycemic (3mM)
Normal (5mM)
Hyperglycemic (8mM)



Biocompatibility
Bulk electrolysis
Live/dead assay
•
Biofilm formation
Basic Components






Anode: 060 Toray Paper electrodes
Fuel: Glucose
Enzyme: Glucose Dehydrogenase
Cofactor: NAD+
Electrocatalyst: Poly(methylene green) (PMG)
Modified polymer:
 Nafion®
 Chitosan
Electrode Preparation
Polymer modification
 Nafion®: Tetrabutylammonium bromide (TBAB)
 Chitosan


Hydrophobic
Deacylation
Chitosan
http://www.global-b2b-network.com/
Co-cast polymer and enzyme onto electrode
Soak electrodes in solution of glucose overnight
Experimental Set-up

3, 5, 8mM glucose fuel
NAD+, pH 7.4 phosphate buffer
Bioanode
V

Glass tube
Fuel Solution



Open circuit potential
(~1000secs)
Linear sweep voltammetry
(<1mV/sec)
Power density equation

P=I*V
+
O-ring
Nafion PEM
4.5cm 2 20% Pt
GDE Cathode
O-ring
Glass tube
Diagram of Icell
Air
Potentiostat
Power Density Test Results
5mM Averages
7e-6
7e-6
6e-6
6e-6
Power Density, Watts/cm2
Power Density, Watts/cm2
3mM Averages
5e-6
4e-6
3e-6
2e-6
1e-6
4e-6
3e-6
2e-6
1e-6
0
0
0
1e-5
2e-5
3e-5
Current Density, Amps/cm
4e-5
2
0
1e-5
Deacylated Chitosan
Chitosan
Nafion
8mM Averages
8e-6
Power Density, Watts/cm2
5e-6
2e-5
3e-5
Current Density, Amps/cm
4e-5
5e-5
2
Average Maximum Power Density* µW/cm2
6e-6
4e-6
2e-6
0
0
1e-5
2e-5
3e-5
Current Density, Amps/cm2
4e-5
5e-5
3mM
5mM
8mM
Chitosan
2.87(±0.21)
2.82(±0.52)
3.32(±0.46)
Deacylated
chitosan
6.04(±3.23)
6.15(±3.51)
7.52(±4.31)
Nafion®
0.28(±0.02)
0.29(±0.02)
0.33(±0.04)
*errors are equal to one standard deviation
Biocompatibility, Bulk Electrolysis
Decreasing current

Possible biofilm
formation
6e-6
5e-6
Current, Amps
Testing
 Bacteria culture
injected
 Hold fuel cell at 0.3V
and monitor current (3
days)

4e-6
3e-6
2e-6
1e-6
0
0.0
5.0e+4
1.0e+5
1.5e+5
Time, seconds
2.0e+5
2.5e+5
Biocompatibility, Live/dead Assay
Live/Dead assay
 Cast polymer with bacteria





Gluconobacter SP33
Origami C4-AW genetically modified E. Coli
Fluorescent nucleic acid stains
FITC filter- live bacteria
TRITC filter- dead bacteria
Live/Dead Assay
Assay showed
biocompatibility
for all polymers.
FITC filter
Chitosan E. coli
Deacylated chitosan
Gluconobacter
Nafion® E. coli
Nafion® Gluconobacter
TRITC filter image
Olympus IX71 fluorescence microscope
Conclusions



Chitosan and Nafion® can immobilize GDH
Chitosan provides higher power and current
densities
Chitosan and Nafion® provide biocompatible
surface material
Future work


Temperature and pH studies
Biocompatible modifications

Impact on current densities
Acknowledgements

National Science Foundation

Saint Louis University

Dr. Minteer

Minteer group
 Kyle Sjöholm
 Dr. Waheed

Rob Arechederra
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