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

Report
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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References
Akers, Moore, Minteer. “Development of Alcohol/O2 Biofuel Cells Using Salt-Extracted Tetrabutylammonium Bromide/Nafion Membranes to Immobilize
Dehydrogenase Enzymes.” Electrochimica Acta 50 (2005): 2521-2525.
Arechederra, Robert, Shelley D. Minteer. “Organelle-based Biofuel Cells: Immobilized Mitochondria on Carbon Paper Electrodes.” Electrochimica Acta 53 (2008):
6698-6703.
Atanassov, Plamen, et al. “Enzymatic Biofuel Cells. The Electrochemical Society Interface (2007).
Beilke, Michael C., et al. “Enzymatic Biofuel Cells.” Micro Fuel Cells Principles and applications. T.S. Zhao. Publisher location: Elsevier, 2009. 179-242. print.
Blackwell, Anne E, et al. “Comparison of Electropolymerized Thiazine Dyes as an Electrocatalyst in Enzymatic Biofuel Cells and Self Powered Sensors.”
Nanoscience and Nanotechnology 9.3 (2009): 1714-21.
Bond, Alan M, et al. “A Role for Electrospray Mass Spectrometry in Electrochemical Studies.” Analytical Chemistry 67(1995):1691-1695.
Cooney, M.J., et al. “Enzyme Catalysed Biofuel Cells.” Energy & Environmental Science 1 (2008): 320-337.
Cox, James A., Thomas J. Gray, “Controlled-Potential Electrolysis of Bulk Solutions at a Modified Electrode: Application to Oxidations of Cysteine, Cystine,
Methionine, and Thiocyanate.” Analytical Chemistry 62(1990): 2742-2744.
Crittenden, Scott R., Christian J. Sund, James J. Sumner. “Mediating Electron Transfer from Bacteria to a Gold Electrode via a Self-Assembled Monolayer.”
Langmuir 22(2006):9473-9476.
Galassetti, Pietro R., et al. “Breath Ethanol and Acetone as Indicators of Serum Glucose Levels: An Initial Report.” Diabetes Technology & Therapeutics 7(2005):115123.
Hu, Qiang, A. Scott Hinman. “A Bulk Electrolysis Raman Spectroelectrochemical Cell Using a Rotating Electrode.” Analytical Chemistry 72(2000): 3233-3235.
Ikeda, Tokuji. “A Novel Electrochemical Approach to the Characterization of Exidoreductase Reactions.” The Chemical Record 4(2004):192-203.
Klotzbach, Tamara L., Michelle Watt, Yasmin Ansari, Shelley D. Minteer. “Improving the microenvironment for enzyme immobilization at electrodes by
hydrophobically modifying chitosan and Nafion® polymers.” Journal of Membrane Science 311(2008):81-88.
“Live/Dead Baclight Bacteria Viability Kitis.” Molecular Probes Inc. 2004.
Mano, Nicolas, “A 280µW cm-2 biofuel cell operating at low glucose concentration.” The Royal Society of Chemistry (2008):2221-2223.
Martin, Georgianna L., Shelley D. Minteer, Michael J. Cooney. “Spatial Distribution of Malate Dehydrogenase in Chitosan Scaffolds.” Applied Materials & Interfaces 1
(2009):367-372.
Minteer, Shelley D., Bor Yann Liaw, Michael J. Cooney. “Enzyme-based biofuel cells.” Current Opinion in Biotechnology 18 (2007):228-234.
Moore, Christine M, et al. “Improving the Environment for Immobilized Dehydrogenase Enzymes by Modifying Nafion with Tetraalkylammonium Bromides.”
Biomacromolecules 5 (2004): 1241-1247.
Subramanyam, Elango, Sidharthan Mohandoss, Hyun-Woung Shin. “Synthesis, Characterization, and Evaluation of Antifouling Polymers of 4Acryloyloxybenzaldehyde with Methyl Methacrylate.” Journal of Applied Polymer Science 112(2009):2741-2749.
Tamaki, T., T. Ito, T. Yamaguchi. “Modelling of Reaction and Diffusion Processes in a High-surface-area Biofuel Cell Electrode Made of Redox Poluymer-grated
Carbon.” Fuel Cells 09 1(2009):37-43.
Wang, J., et al. “The effects of amorphous carbon films deposited on polyethylene terephthalate on bacterial adhesion.” Biomaterials 25(2004):3163-3170.
Heikkila, O., N Lundbom, M Timonen, P-H Groop, S Heikkinen, S Makimattila. “Hyperglycaemia is associated with changes in the regional concentrations of glucose
and myo-inositol within the brain.” Diabetologia 52(2009):534-540.
Gupta, Sandeep, Eugene Chough, Jennifer Daley, Peter Oates, Keith Tornheim, Neil B. Ruderman, and John F. Keaney Jr. “Hyperglycemia increases endothelial
superoxide that impairs smooth muscle cell Na+-K+-ATPase activity.” Am J Physiol Cell Physiol 282(2002): C560-C566.
Mason RM,Thomas G, Davies M. “Proteoglycan synthesis by human mesangial cells is depressed by hyperglycemic glucose concentrations.” Biochemical society
transactions 2(1992): 96
Xiaoli, Ma, Yao Zihua, Shi Dagang. “Preparation and characterization of porous chitosan membranes and the localization of the activity of urease immobilized on it by
SEM and X-ray microanalysis.” Chemical Journal on Internet 7(2005): 45.

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