Applied Microbiology and Biotechnology

, Volume 89, Issue 5, pp 1315–1322 | Cite as

Strategies to extend the lifetime of bioelectrochemical enzyme electrodes for biosensing and biofuel cell applications

  • Stefanie Rubenwolf
  • Sven Kerzenmacher
  • Roland Zengerle
  • Felix von Stetten


Enzymes are powerful catalysts for biosensor and biofuel cell electrodes due to their unique substrate specificity. This specificity is defined by the amino acid chain's complex three-dimensional structure based on non-covalent forces, being also responsible for the very limited enzyme lifetime of days to weeks. Many electrochemical applications, however, would benefit from lifetimes over months to years. This mini-review provides a critical overview of strategies and ideas dealing with the problem of short enzyme lifetime, which limits the overall lifetime of bioelectrochemical electrodes. The most common approaches aim to stabilize the enzyme itself. Various immobilization techniques have been used to reduce flexibility of the amino acid chain by introducing covalent or non-covalent binding forces to external molecules. The enzyme can also be stabilized using genetic engineering methods to increase the binding forces within the protein or by optimizing the environment in order to reduce destabilizing interactions. In contrast, renewing the inactivated catalyst decouples overall system lifetime from the limited enzyme lifetime and thereby promises theoretically unlimited electrode lifetimes. Active catalyst can be supplied by exchanging the electrolyte repeatedly. Alternatively, integrated microorganisms can display the enzymes on their surface or secrete them to the electrolyte, allowing unattended power supply for long-term applications.


Enzyme inactivation Biofuel cell Biosensor Amino acid replacement Immobilization Self-regeneration 



Financial support by the German Research Foundation (DFG) through the Ph.D. program “Micro Energy Harvesting” (GRK 1322) and the German Ministry of Education and Research (BMBF, Projektträger Jülich) under the program “Bioenergie2021” (Grant No. 03SF0382) is gratefully acknowledged.


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Stefanie Rubenwolf
    • 1
  • Sven Kerzenmacher
    • 1
  • Roland Zengerle
    • 1
    • 2
  • Felix von Stetten
    • 1
  1. 1.Laboratory for MEMS Applications, Department of Microsystems Engineering-IMTEKUniversity of FreiburgFreiburgGermany
  2. 2.Centre for Biological Signalling Studies–BiossAlbert-Ludwigs-Universität FreiburgFreiburgGermany

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