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A Bioelectrocatalysis Method for the Kinetic Measurement of Thermal Inactivation of a Redox Enzyme, Bilirubin Oxidase

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Abstract

The thermal stability of a redox enzyme, bilirubin oxidase (BOD), has been quantitatively evaluated by measuring the inactivation kinetics of BOD at several temperatures. The enzyme activity is directly related to the mediated bioelectrocatalytic current for the BOD-catalyzed reduction of O2. Thus, the inactivation process is measured by the time-dependent decrease in the bioelectrocatalytic current. The results reveal that the inactivation obeys first-order kinetics, whose rate constants (k) are determined at pH 7.0 and at 50–70°C. The half life of BOD activity, calculated from the k value at 50°C is 114 min, which is in harmony with the thermal-stability data given in a catalog by Amano Enzyme Inc. The bioelectrocatalysis method allows in situ measurements of the inactivation kinetics in the period of a few minutes at relatively high temperatures. The rate constants show a large temperature dependence, leading to a large Arrhenius activation energy (EA) of 221 kJ mol-1. The activation Gibbs energy (ΔG≠), activation enthalpy (ΔH≠), and activation entropy (ΔS≠) are also determined.

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References

  1. “Kouso Kougaku (Enzyme Technology, in Japanese)”, ed. S. Fukui, I. Chibata, and S. Suzuki, 1982, Tokyo-kagaku-dojin, Tokyo.

    Google Scholar 

  2. K. Kano and T. Ikeda, Bunseki (in Japanese), 2003, 576.

    Google Scholar 

  3. K. Kano and T. Ikeda, Electrochemistry, 2003, 71, 86.

    Article  CAS  Google Scholar 

  4. “Baiodenkikagaku no Jissai—Baiosensa—Baiodenchi no Jitsuyou Tenkai (Practical Bioelectrochemistry—Recent Developments in Biosensors & Biofuel Cells, in Japanese), ed. T. Ikeda, 1982, CMC Publishing, Tokyo.

    Google Scholar 

  5. S. C. Barton, J. Gallaway, and P. Atanasov, Chem. Rev., 2004, 104, 4867.

    Article  CAS  Google Scholar 

  6. T. Hibi and Y. Nishiya, p.101 in Ref. 4.

  7. Thermal stability of enzymes is expressed as such in catalogs of enzyme companies. For example, the thermal stability of BOD appears in Amano Enzyme Catalog, 18 (http://www.amano-enzyme.co.jp/jp/productinfo/medical 03.html).

  8. T. Ikeda, Chem. Rec., 2004, 4, 192.

    Article  CAS  Google Scholar 

  9. S. Tsujimura, H. Tatsumi, J. Ogawa, S. Shimizu, K. Kano, and T. Ikeda, J. Electroanal. Chem., 2001, 496, 69.

    Article  CAS  Google Scholar 

  10. S. Tsujimura, M. Fujita, H. Tatsumi, K. Kano, and T. Ikeda, Phys. Chem. Chem. Phys., 2001, 3, 1331.

    Article  CAS  Google Scholar 

  11. S. Tsujimura, K. Kano, and T. Ikeda, Electrochemistry, 2002, 70, 940.

    Article  CAS  Google Scholar 

  12. A. Shimizu, J. H. Kwon, T. Satoh, N. Sakurai, T. Sakurai, S. Yamaguchi, and T. Samejima, Biochemistry, 1999, 38, 3034.

    Article  CAS  Google Scholar 

  13. T. Nakagawa, S. Tsujimura, K. Kano, and T. Ikeda, Chem. Lett., 2003, 32, 54.

    Article  CAS  Google Scholar 

  14. A. Shimizu, J. H. Kwon, T. Satoh, N. Sakurai, T. Sakurai, S. Yamaguchi, and T. Samejima, J. Biochem., 1999, 125, 662.

    Article  CAS  Google Scholar 

  15. S. Tsujimura, M. Kawahara, T. Nakagawa, K. Kano, and T. Ikeda, Electrochem. Commun., 2003, 5, 138.

    Article  CAS  Google Scholar 

  16. R. Matsumoto, K. Kano, and T. Ikeda, J. Electroanal. Chem., 2002, 535, 37.

    Article  CAS  Google Scholar 

  17. T. Ikeda, H. Tatsumi, H. Katano, and K. Kano, p. 38 in Ref. 4.

  18. A. J. Bard and L. R. Faulkner, “Electrochemical Methods: Fundamentals and Applications”, 1982, John Wiley and Sons, New York, Chichester, Brisbane, Toronto, 457.

    Google Scholar 

  19. T. Ikeda, H. Tatsumi, H. Katano, and K. Kano, p. 43 in Ref. 4.

  20. K. Hiromi, “Kouso Hannou Kaiseki no Jissai (Practice in Kinetic Analysis of Enzyme Reactions, in Japanese)”, 1982, Koudansya, Tokyo, 54.

    Google Scholar 

  21. T. K. Harris and V. L. Davidson, Biochem. J., 1994, 303, 141.

    Article  CAS  Google Scholar 

  22. A. N. Eremin, D. I. Metelitsa, Zh. F. Shishko, R. V. Mikhailova, M. I. Iasenko, and A. G. Lobanok, Prikl. Biokhim. Mikrobiol., 2001, 37, 678.

    CAS  PubMed  Google Scholar 

  23. M. D. Gouda, S. A. Singh, A. G. Rao, M. S. Thakur, and N. G. Karanth, J. Biol. Chem., 2003, 278, 24324.

    Article  CAS  Google Scholar 

  24. D. N. Rani and T. E. Abraham, Appl. Biochem. Biotechnol., 2006, 128, 215.

    Article  CAS  Google Scholar 

  25. S. Koikeda, K. Ando, H. Kaji, T. Inoue, S. Murao, K. Takeuchi, and T. Samejima, J. Biol. Chem., 1993, 268, 18801.

    Article  CAS  Google Scholar 

  26. A. Shimizu, J. H. Kwon, T. Sasaki, T. Satoh, N. Sakurai, T. Sakurai, S. Yamaguchi, and T. Samejima, Biochemistry, 1999, 38, 3034.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Bioscience, Fukui Prefectural University, Fukui, 910-1195, Japan

    Tokuji Ikeda, Hirosuke Tatsumi, Hajime Katano, Mizue Wanibuchi & Takao Hibi

  2. Toyota Central R & D Labs Inc., Yokomichi, Nagakute, Aichi, 480-1192, Japan

    Tsutomu Kajino

  3. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo, Kyoto, 606-8502, Japan

    Mizue Wanibuchi

Authors
  1. Tokuji Ikeda
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  2. Hirosuke Tatsumi
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  3. Hajime Katano
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  4. Mizue Wanibuchi
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  5. Takao Hibi
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  6. Tsutomu Kajino
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Ikeda, T., Tatsumi, H., Katano, H. et al. A Bioelectrocatalysis Method for the Kinetic Measurement of Thermal Inactivation of a Redox Enzyme, Bilirubin Oxidase. ANAL. SCI. 24, 237–241 (2008). https://doi.org/10.2116/analsci.24.237

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  • DOI: https://doi.org/10.2116/analsci.24.237

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