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DOI: 10.1002/celc.201402162
The utilization of carbon felt as the conductive material for the helped in the immobilization of a larger quantity of enzyme.
construction of a monosaccharide-based coin-cell biobattery is Cathodic and anodic currents with carbon felt electrodes
explored. Anthracene-modified carbon nanotubes were used showed a three-fold and twofold increase, respectively, relative
at the positive electrode to preferentially orientate laccase for to the currents obtained with Toray paper materials. Bioelec-
direct electron transfer during O2 reduction. A ferrocene-modi- trodes were assembled in a commercial coin-cell battery
fied poly(ethylenimine) redox polymer was used to electrically casing and were tested as possible biobatteries. This work
communicate with either glucose oxidase or fructose dehydro- presents the first time in which a traditional battery design is
genase at the negative electrode. The use of carbon felt used for the performance evaluation of different biobatteries.
1. Introduction
Interest in biofuel cells has grown dramatically over the last do not solve the substrate diffusion issue as shown in recent
decade.[1–3] By immobilizing oxidizing enzymes at the anode papers.[10]
and reducing enzymes at the cathode, a voltage can be gener- Much of the early work with high current density biofuel
ated that, if stacked, is sufficient to operate a wide range of cells has focused on the use of redox polymers to immobilize
useful electronics.[4–6] However, despite a relatively large theo- an enzyme onto the surface of an electrode. In such electrode
retical potential, many biofuel cells are limited by the amount films, redox mediators help shuttle electrons from the active
of current that can be generated due to limitations in diffusion site of the enzyme to the electrode surface. Mediated electron
of either the substrate or the electrons through the electrode transfer is preferable for enzymes such as glucose oxidase, as
material. By either vigorously stirring the solution or using a ro- the active site resides in the interior of the enzyme, and this
tating disk electrode, many of the substrate diffusion limita- makes it inaccessible for direct electron transfer to the elec-
tions can be overcome. However, it should be noted that nei- trode surface.[11–13]
ther of these options is an applicable solution to the problem Recent research has shown that redox polymers based on
of substrate diffusion, but rather a means of determining theo- ferrocene-modified linear poly(ethylenimine) (LPEI) form hydro-
retical limitations of various biofuel-cell materials.[7] A practical gels if crosslinked in the presence of an enzyme on the surface
solution to the problem of electron diffusion is the use of of an electrode. The ferrocene–LPEI polymers interact favorably
carbon nanotubes (CNTs) and various types of mesoporous with enzymes such as glucose oxidase through complexation
materials as a means of dramatically increasing the amount of between positive charges along the polymer backbone and
real surface area and decreasing the diffusional distances be- the negatively charged enzyme surface, which allows for cur-
tween redox species (redox mediators or active centers of en- rent densities as high as 2 mA cm 2 in the presence of glucose
zymes) and electrode surfaces.[8, 9] Nonetheless, these materials on planar surfaces.[14]
Unlike glucose oxidase, laccase contains a T1 Cu electron
relay that is readily accessible from the enzyme’s surface, and
it can shuttle the electrons to the active site. Work has been
[a] Dr. F. Giroud, Prof. S. D. Minteer
Department of Chemistry, University of Utah done to show that anthracene-modified multiwalled CNTs (An-
315 S 1400 E Rm 2020 MWCNTs) can bind to the active site of laccase to control the
Salt Lake City, Utah 84112 (USA) orientation of the enzyme.[15, 16] The use of An-MWCNTs to im-
Phone: 1-801-587-8325
mobilize laccase allows for a very efficient means of direct elec-
E-mail: minteer@chem.utah.edu
tron transfer, which can result in cathodic current densities as
[b] Dr. D. P. Hickey, Prof. D. T. Glatzhofer
Department of Chemistry and Biochemistry high as 1.84 mA cm 2 in the presence of O2.[17]
University of Oklahoma, 101 Stephenson Parkway In this report, we study the use of carbon felt (CF) electrodes
Norman, Oklahoma 73019 (USA) as the conductive support material in place of Toray paper (TP)
[c] Prof. D. W. Schmidtke electrodes for the construction of enzyme-based bioelectrodes.
University of Oklahoma Biomedical Engineering Center
For this purpose, we utilize a combination of ferrocene-modi-
and School of Chemical, Biological, Materials Engineering
University of Oklahoma, 100 East Boyd fied linear poly(ethylenimine) (Fc-C6-LPEI) and glucose oxidase
Norman, Oklahoma 73019 (USA) (GOx) or fructose dehydrogenase (FDH) at the bioanode,
An invited contribution to a Special Issue on Biofuel Cells whereas an ink of An-MWCNTs and laccase is used at the cath-
2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim ChemElectroChem 2014, 1, 1880 – 1885 1880
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ode. Finally, we investigate a new approach for the characteri- ductase (17.2 mL, 13 mg mL 1) was added to solution 1, and the so-
zation of biobatteries by sealing the bioanode and the bioca- lution was mixed by vortexing for 1 min. Finally, a solution of 4.3 %
thode in a commercial, self-contained coin-cell design to con- EGDGE (6.44 mL, EGDGE/water = 2:45 v/v) was added to the previ-
ous mixture. The resulting solution was mixed thoroughly. The TP
struct a monosaccharide/dioxygen biobattery. The coin-cell
and CF electrodes were modified by dropcasting from the final so-
battery is tested with glucose and fructose for comparison.
lution (25 and 50 mL, respectively), and the resulting films were al-
This work presents an attractive method to help standardize lowed to dry in open air at room temperature overnight to ensure
the testing done for the performance evaluation of biobatter- crosslinking. In the case of the CF electrodes, the felt was thor-
ies in development. oughly soaked in deionized water to help the hydrogel to diffuse
inside the pores.
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rocene in the Fc-C6-LPEI is originally in its reduced state, experi- 2.2. Half-Cell Bioanode Characterization: Carbon Material
ments were recorded after a second polarization of the biobattery Studies
from OCV to 5 mV to guarantee that the Fc-C6-LPEI was in its oxi-
dized state before characterization in the presence of fuel. The fuel
Bioelectrocatalysis of the bioanodes was tested under different
solution was flushed through the battery to allow the solution
conditions at 25 8C in 50 mm citrate buffer at pH 5.0. To in-
inside the casing to be constant between experiments (namely,
100 mm substrate). The OCV was allowed to stabilize until it crease the current at the electrodes, CNT dispersions were
reached a steady state, and linear scan voltammetry (LSV) was per- added to the hydrogel modification. Different CNT dispersion
formed in the presence of 100 mm fuel solution.
2. Results
2.1. Half-Cell Biocathode Characterization: Carbon Material
Studies
Two different electrode materials were used in this study to
compare the bioelectrocatalytic currents obtained from the
oxygen reduction reaction from laccase at An-MWCNTs. TP and
CF electrodes were modified on a single side with the same
amount of biocathode ink. Figure 1 depicts the bioelectrocata-
lytic current obtained at these electrodes immersed in 50 mm
citrate buffer, pH 4.5. The larger porosity (from 0.94 to
0.98)[21, 22] and greater thickness (3.18 mm) of the CF material
relative to the same values of the TP material (0.78 and
0.19 mm, specification sheet) provided a higher capacity load-
ing for incorporation of the bioink into the conductive materi-
al. Current densities were higher on the CF electrodes with
a twofold and 1.5-fold increase relative to those on the TP
electrodes in the presence of dissolved and saturated oxygen,
respectively (TP: 115.4 10.0 and 299.6 33.5 mA cm 2 for dis-
solved and saturated O2, respectively; CF: 242.1 15.1 and
458.6 44.4 mA cm 2 for dissolved and saturated O2, respective-
ly). To estimate the benefit of the CF material on the bioink ca-
pacity loading at the electrode, the biocathode ink was depos-
ited on both sides of the CF material and the volume used
was increased by a factor of two and four on each side for
each electrode. Bioelectrocatalysis of oxygen reduction was
performed in quiescent solution (dissolved O2) and under satu-
rated oxygen conditions. All the current densities are reported
in Table 1. The resulting currents were increased two- and
threefold relative to those of the single-side-modified CF
electrodes.
Finally, both working conditions and stability experiments Figure 1. Representative cyclic voltammograms of laccase/An-MWCNT im-
were performed at pH 5.0 to assess the electrocatalytic current mobilized on TP electrodes (top) and CF electrodes (bottom) soaked in
50 mm citrate buffer at pH 4.5 in dissolved O2 (c) or under pure O2 bub-
that could be obtained in the biofuel cell experiment running bling (a) conditions recorded at 1 mV s 1.
under the same conditions. Upon first testing by CV, the cur-
rent recorded from An-MWCNT/
laccase biocathodes at + 300 mV
versus SCE was 682.3
Table 1. Comparison of the catalytic current of different laccase/An-MWCNT-modified CF electrodes depending
66.3 mA cm 2. The current was on the bioink loading and oxygen concentration.[a]
monitored continuously by CV
and amperometric studies for Conditions Current density [mA cm 2]
Single side modification Two side modification
a period of 7 days. As shown in
Twofold increase in volume Fourfold increase in volume
Figure 2, electrocatalytic currents
of the biocathodes decreased dissolved O2 254.5 13.6 546.9 32.1 (2.3) 774.4 43.7 (3.0)
saturated O2 518.8 54.7 1180.0 110.3 (2.3) 1830.0 78.5 (3.5)
progressively to reach 65 % of
the original currents after 7 days [a] Cyclic voltammograms were recorded at 1 mV s 1 in 50 mm citrate buffer, pH 4.5. Current densities were
taken at 300 mV vs. SCE.
of continuous operation.
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3. Conclusions
The use of CF electrodes allows a larger quantity of bioelectro-
catalyst to be immobilized at an electrode relative to that al-
lowed by conventional TP. These CF electrodes can be sealed
in a coin-cell casing design to form a biobattery. By using lac-
case at the biocathode and fructose dehydrogenase or glucose
oxidase at the anode, we were able to demonstrate the proof
of concept of this design working with a relatively small
volume of fuel solution. Upon combining a bioanode and a bio-
cathode into a biobattery, there are many new issues that
need to be addressed from this new design, such as the small
volume required and the production of hydrogen peroxide
within the battery. Further experiments will be performed to
Figure 5. Polarization (left) and power (right) curves obtained for laccase/ increase the open-circuit voltage of the battery, which drasti-
GOx or laccase/FDH coin-cell biobatteries in the absence or presence of sub- cally limits the performances of the devices. Nonetheless, we
strate at 1 mV s 1. All the electrodes were made with CF. Electrolyte: 50 mm
have shown that commercial battery casings are suitable to
citrate, pH 5.0 at 25 8C.
characterize the performances of two different biobatteries,
and this could be the starting point for a more general stand-
the biobattery at high current densities. These results are in ardization testing procedure for all the biobatteries in
agreement with the previous results obtained from the CV ex- development.
periments we reported for the two enzymes by using a FcMe4-
C3-LPEI.[24] The performances obtained were not as high as
those of other biobattery designs found in the literature, but Acknowledgements
they could be improved by incorporating a redox polymer
with a lower oxidation potential or by optimizing the bioca- The authors would like to acknowledge the National Science
thode ink and bioanode hydrogel loading. Furthermore, to the Foundation (NSF) (Grant no. 1057597) for funding. The authors
authors’ best knowledge, this is the first example in which would also like to acknowledge the NSF CBET 0967988 program
a biobattery combines two bioelectrodes. Other biobattery de- for funding.
signs involve the use of hybrid technology by combining an
enzymatic bioelectrode and an abiotic electrode. The Minteer Keywords: biobatteries · biofuel cells · carbohydrates ·
group reported several biobattery designs based on nicotin- conducting materials · enzymes
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