Section A: Enzyme-Based Biofuel Cells

Engineered Pyranose 2-Oxidase: Efficiently Turning Sugars into Electrical Energy

  1. Oliver Spadiut1,
  2. Dagmar Brugger2,
  3. Vasile Coman3,
  4. Dietmar Haltrich2,
  5. Lo Gorton3

Article first published online: 22 FEB 2010

DOI: 10.1002/elan.200980015

Issue

Electroanalysis

Electroanalysis

Special Issue: Biofuel Cells

Volume 22, Issue 7-8, pages 813–820, April 2010

Additional Information

How to Cite

Spadiut, O., Brugger, D., Coman, V., Haltrich, D. and Gorton, L. (2010), Engineered Pyranose 2-Oxidase: Efficiently Turning Sugars into Electrical Energy. Electroanalysis, 22: 813–820. doi: 10.1002/elan.200980015

Author Information

  1. 1

    School of Biotechnology, KTH, Albanova University Centre, SE-106 91 Stockholm, Sweden

  2. 2

    Department of Food Sciences and Technology, BOKU – University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austria

  3. 3

    Department of Analytical Chemistry, Lund University, P. O. Box 124, SE-22100 Lund, Sweden

Publication History

  1. Issue published online: 1 APR 2010
  2. Article first published online: 22 FEB 2010
  3. Manuscript Accepted: 19 OCT 2009
  4. Manuscript Received: 16 OCT 2009

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Keywords:

  • Pyranose 2-oxidase;
  • Sugar oxidoreductases;
  • Rational protein design;
  • Enzymatic biofuel cell;
  • Mediated electron transfer;
  • Osmium r1edox polymer;
  • 1,4-Benzoquinone;
  • Fuel cells

Abstract

Due to the recent interest in enzymatic biofuel cells (BFCs), sugar oxidizing enzymes other than the commonly used glucose oxidase are gaining more importance as possible bioelements of implantable microscale-devices, which can, for example, be used in biosensors and pacemakers. In this study we used rational and semi-rational protein design to improve the catalytic activity of the enzyme pyranose 2-oxidase (P2Ox) with its alternative soluble electron acceptors 1,4-benzoquinone and ferricenium ion, which can serve as electron mediators, to possibly boost the power output of enzymatic BFCs. Using a screening assay based on 96-well plates, we identified the variant H450G, which showed lower KM and higher kcat values for both 1,4-benzoquinone and ferricenium ion compared to the wild-type enzyme, when either D-glucose or D-galactose were used as saturating electron donors. Besides this variant, we analyzed the variants V546C and T169G/V546C for their possible application in enzymatic BFCs. The results obtained in homogeneous solution were compared with those obtained when P2Ox was immobilized on the surface of graphite electrodes and either “wired” to an osmium redox polymer or using soluble 1,4-benzoquinone as mediator. According to the spectrophotometrically determined kinetic constants, the possible energy output, measured in flow injection analysis experiments with these variants, increased up to 4-fold compared to systems employing the wild-type enzyme.

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