Skip to main content
Log in

Prolongation of electrode lifetime in biofuel cells by periodic enzyme renewal

  • Bioenergy and Biofuels
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Enzymatically catalyzed biofuel cells show unique specificity and promise high power densities, but suffer from a limited lifetime due to enzyme deactivation. In the present work, we demonstrate a novel concept to extend the lifetime of a laccase-catalyzed oxygen reduction cathode in which we decouple the electrode lifetime from the limited enzyme lifetime by a regular resupply of fresh enzymes. Thereto, the adsorption behavior of laccase from Trametes versicolor to buckypaper electrode material, as well as its time-dependent deactivation characteristics, has been investigated. Laccase shows a Langmuir-type adsorption to the carbon nanotube-based buckypaper electrodes, with a mean residence time of 2 days per molecule. In a citrate buffer of pH 5, laccase does not show any deactivation at room temperature for 2 days and exhibits a half-life of 9 days. In a long-term experiment, the laccase electrodes were operated at a constant galvanostatic load. The laccase-containing catholyte was periodically exchanged against a freshly prepared one every second day to provide sufficient active enzymes in the catholyte for the replacement of desorbed inactive enzymes. Compared to a corresponding control experiment without catholyte exchange, this procedure resulted in a 2.5 times longer cathode lifetime of 19 ± 9 days in which the electrode showed a potential above 0.744 V vs. normal hydrogen electrode at 110 μA cm−2. This clearly indicates the successful exchange of molecules by desorption and re-adsorption and is a first step toward the realization of a self-regenerating enzymatic biofuel cell in which enzyme-producing microorganisms are integrated into the electrode to continuously resupply fresh enzymes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Antiohos D, Moulton SE, Minett AI, Wallace GG, Chen J (2010) Electrochemical investigation of carbon nanotube nanoweb architecture in biological media. Electrochem Commun 12:1471–1474

    Article  CAS  Google Scholar 

  • Atanasov P, Yang S, Salehi C, Ghindilis AL, Wilkins E, Schade D (1997) Implantation of a refillable glucose monitoring-telemetry device. Biosens Bioelectron 12:669–680

    Article  CAS  Google Scholar 

  • Atkins P, de Paula J (2006) Atkin’s physical chemistry, 8th edn. Oxford University Press, Oxford

    Google Scholar 

  • Binjamin G, Chen T, Heller A (2001) Sources of instability of ‘wired’ enzyme anodes in serum: urate and transition metal ions. J Electroanal Chem 500:604–6011

    Article  Google Scholar 

  • Blanford CF, Heath RS, Armstrong FA (2007) A stable electrode for high-potential, electrocatalytic O2 reduction based on rational attachment of a blue copper oxidase to a graphite surface. Chem Commun 17:1710–1712

    Article  Google Scholar 

  • Chase HA (1984) Prediction of the performance of preparative affinity-chromatography. J Chromatogr 297:179–202

    Article  CAS  Google Scholar 

  • Cinquin P, Gondran C, Giroud F, Mazabrard S, Pellissier A, Boucher F, Alcaraz JP, Gorgy K, Lenouvel F, Mathé S, Porcu P, Cosnier S (2010) A glucose biofuel cell implanted in rats. PLoS One 5:e10476

    Article  Google Scholar 

  • Coll PM, Perez P, Villar E, Shnyrov VL (1994) Domain-structure of laccase-I from the lignin-degrading basidiomycete PM1 revealed by differential scanning calorimetry. Biochem Mol Biol Int 34:1091–1098

    CAS  Google Scholar 

  • Fahraeus G, Reinhammar B (1967) Large scale production and purification of laccase from cultures of fungus Polyporus versicolor and some properties of laccase A. Acta Chem Scand 21:2367–2378

    Article  CAS  Google Scholar 

  • Fishilevich S, Amir L, Fridman Y, Aharoni A, Alfonta L (2009) Surface display of redox enzymes in microbial fuel cells. J Am Chem Soc 131:12052–12053

    Article  CAS  Google Scholar 

  • Gellett W, Schumacher J, Kesmez M, Le D, Minteer SD (2010) High current density air-breathing laccase biocathode. J Electrochem Soc 157:B557–B562

    Article  CAS  Google Scholar 

  • Gianazza E, Crawford J, Miller I (2007) Detecting oxidative post-translational modifications in proteins. Amino Acids 33:51–56

    Article  CAS  Google Scholar 

  • Horozova E, Dimcheva N (2004) Kinetic study of catalase adsorption on disperse carbonaceous matrices. Cent Eur J Chem 2:627–637

    Article  CAS  Google Scholar 

  • Hussein L, Rubenwolf S, von Stetten F, Urban G, Zengerle R, Krueger M, Kerzenmacher S (2011a) A highly efficient buckypaper-based electrode material for mediatorless laccase-catalyzed dioxygen reduction. Biosens Bioelectron 26:4133–4138

    Article  CAS  Google Scholar 

  • Hussein L, Urban G, Krüger M (2011b) Fabrication and characterization of buckypaper-based nanostructured electrodes as a novel material for biofuel cell applications. Phys Chem Chem Phys 13:5831–5839

    Article  CAS  Google Scholar 

  • Karpovich DS, Blanchard GJ (1994) Direct measurement of the adsorption-kinetics of alkanethiolate self-assembled monolayers on a microcrystalline gold surface. Langmuir 10:3315–3322

    Article  CAS  Google Scholar 

  • Kerzenmacher S, Mutschler K, Kräling U, Baumer H, Ducrée J, Zengerle R, von Stetten F (2009) A complete testing environment for the automated parallel performance characterization of biofuel cells: design, validation, and application. J Appl Electrochem 39:1477–1485

    Article  CAS  Google Scholar 

  • Kiiskinen LL, Palonen H, Linder M, Viikari L, Kruus K (2004) Laccase from Melanocarpus albomyces binds effectively to cellulose. FEBS Lett 576:251–255

    Article  CAS  Google Scholar 

  • Kloke A, Rubenwolf S, Bücking C, Gescher J, Kerzenmacher S, Zengerle R, von Stetten F (2010) A versatile miniature bioreactor and its application to bioelectrochemistry. Biosens Bioelectron 25:2559–2565

    Article  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during assembly of head of bacteriophage-T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Langmuir I (1918) The adsorption of gases on plane surfaces of glass, mica and platinum. J Am Chem Soc 40:1361–1403

    Article  CAS  Google Scholar 

  • Lee CW, Gray HB, Anson FC, Malmstrom BG (1984) Catalysis of the reduction of dioxygen at graphite-electrodes coated with fungal laccase-A. J Electroanal Chem 172:289–300

    Article  CAS  Google Scholar 

  • Malmstrom BG, Kimmel JR, Smith EL (1959) Amino acid composition and amino-terminal sequence of yeast enolase. J Biol Chem 234:1108–1111

    CAS  Google Scholar 

  • Piontek K, Antorin M, Choinowski T (2002) Crystal structure of a laccase from the fungus Trametes versicolor at 1.90-angstrom resolution containing a full complement of coppers. J Biol Chem 277:37663–37669

    Article  CAS  Google Scholar 

  • Rubenwolf S, Strohmeier O, Kloke A, Kerzenmacher S, Zengerle R, von Stetten F (2010) Carbon electrodes for direct electron transfer type laccase cathodes investigated by current density–cathode potential behavior. Biosens Bioelectron 26:841–845

    Article  CAS  Google Scholar 

  • Rubenwolf S, Kerzenmacher S, Zengerle R, von Stetten F (2011) Strategies to extend the lifetime of bioelectrochemical enzyme electrodes for biosensing and biofuel cell applications. Appl Microbiol Biotechnol 89:1315–1322

    Article  CAS  Google Scholar 

  • Sadana A (1988) Enzyme deactivation. Biotechnol Adv 6:349–446

    Article  CAS  Google Scholar 

  • Szczupak A, Kol-Kalman D, Alfonta L (2012) A hybrid biocathodes: surface display of O2 reducing enzymes for microbial fuel cell applications. Chem Commun 48:49–51

    Article  CAS  Google Scholar 

  • Tarasevich MR, Bogdanovskaya VA, Kuznetsova LN (2001) Bioelectrocatalytic reduction of oxygen in the presence of laccase adsorbed on carbon electrodes. Russ J Electrochem 37:833–837

    Article  CAS  Google Scholar 

  • Warburg O, Christian W (1941) Isolierung und Kristallisierung des Gärungsferments Enolase. Biochem Z 310:384–421

    Google Scholar 

  • Yamaguchi M, Nakano A, Taniyama T (2008) Yeast transformant-based glucose biosensor for implantable application. Sens Mater 20:131–141

    CAS  Google Scholar 

  • Yang S, Atanasov P, Wilkuns E (1997) Development of a dual glucose–oxygen sensor system for continuous in vivo monitoring. J Clin Eng 22:55–63

    Google Scholar 

  • Yaropolov AI, Skorobogatko OV, Vartanov SS, Varfolomeyev SD (1994) Laccase—properties, catalytic mechanism, and applicability. Appl Biochem Biotechnol 49:257–280

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge financial support by the German Research Foundation (DFG) through the PhD program “Micro Energy Harvesting” (GRK 1322) and the German Ministry of Education and Research (BMBF) under the program Bioenergie2021 (grant no. 03SF0382). Furthermore, we would like to thank Dr. Daniel Wohlwend (Laboratory for Biochemistry) for his assistance in understanding laccase unfolding.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to S. Kerzenmacher.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rubenwolf, S., Sané, S., Hussein, L. et al. Prolongation of electrode lifetime in biofuel cells by periodic enzyme renewal. Appl Microbiol Biotechnol 96, 841–849 (2012). https://doi.org/10.1007/s00253-012-4374-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-012-4374-8

Keywords

Navigation