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Applied and Environmental Microbiology, December 2008, p. 7329-7337, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.00177-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Microbial Biofilm Voltammetry: Direct Electrochemical Characterization of Catalytic Electrode-Attached Biofilms{triangledown} ,{dagger}

Enrico Marsili,1 Janet B. Rollefson,2 Daniel B. Baron,1 Raymond M. Hozalski,3 and Daniel R. Bond1,4*

BioTechnology Institute, University of Minnesota, St. Paul, Minnesota 55108,1 Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455,2 Department of Civil Engineering, University of Minnesota, Minneapolis, Minnesota 55455,3 Department of Microbiology, University of Minnesota, Minneapolis, Minnesota 554554

Received 18 January 2008/ Accepted 21 September 2008

While electrochemical characterization of enzymes immobilized on electrodes has become common, there is still a need for reliable quantitative methods for study of electron transfer between living cells and conductive surfaces. This work describes growth of thin (<20 µm) Geobacter sulfurreducens biofilms on polished glassy carbon electrodes, using stirred three-electrode anaerobic bioreactors controlled by potentiostats and nondestructive voltammetry techniques for characterization of viable biofilms. Routine in vivo analysis of electron transfer between bacterial cells and electrodes was performed, providing insight into the main redox-active species participating in electron transfer to electrodes. At low scan rates, cyclic voltammetry revealed catalytic electron transfer between cells and the electrode, similar to what has been observed for pure enzymes attached to electrodes under continuous turnover conditions. Differential pulse voltammetry and electrochemical impedance spectroscopy also revealed features that were consistent with electron transfer being mediated by an adsorbed catalyst. Multiple redox-active species were detected, revealing complexity at the outer surfaces of this bacterium. These techniques provide the basis for cataloging quantifiable, defined electron transfer phenotypes as a function of potential, electrode material, growth phase, and culture conditions and provide a framework for comparisons with other species or communities.

* Corresponding author. Mailing address: BioTechnology Institute, University of Minnesota, 140 Gortner Laboratory, 1479 Gortner Ave., St. Paul, MN 55108. Phone: (612) 624-8619. Fax: (612) 625-1700. E-mail: dbond{at}umn.edu

{triangledown} Published ahead of print on 10 October 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, December 2008, p. 7329-7337, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.00177-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Li, Q., Seffernick, J. L., Sadowsky, M. J., Wackett, L. P. (2009). Thermostable Cyanuric Acid Hydrolase from Moorella thermoacetica ATCC 39073. Appl. Environ. Microbiol. 75: 6986-6991 [Abstract] [Full Text]  
  • Baron, D., LaBelle, E., Coursolle, D., Gralnick, J. A., Bond, D. R. (2009). Electrochemical Measurement of Electron Transfer Kinetics by Shewanella oneidensis MR-1. J. Biol. Chem. 284: 28865-28873 [Abstract] [Full Text]  
  • Rollefson, J. B., Levar, C. E., Bond, D. R. (2009). Identification of Genes Involved in Biofilm Formation and Respiration via Mini-Himar Transposon Mutagenesis of Geobacter sulfurreducens. J. Bacteriol. 191: 4207-4217 [Abstract] [Full Text]  


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