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Electrochemical studies on nitrite reductase towards a biosensor

M Scharf1, C Moreno, C Costa

  • 1Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Portugal.

Biochemical and Biophysical Research Communications
|April 26, 1995
PubMed
Summary

A c-type hexaheme nitrite reductase (NiR) from Desulfovibrio desulfuricans shows promise for biosensor development. This enzyme catalyzes nitrite reduction and exhibits Michaelis-Menten kinetics when immobilized on an electrode, enabling nitrite detection.

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Area of Science:

  • Biochemistry
  • Electrochemistry
  • Enzyme catalysis

Background:

  • Nitrite reductase (NiR) is a c-type hexaheme enzyme.
  • Desulfovibrio desulfuricans (Dd) NiR catalyzes the six-electron reduction of nitrite to ammonia.
  • Previous studies indicated a simple electrocatalytic mechanism for this enzyme.

Purpose of the Study:

  • To investigate the potential of Dd NiR as a biosensor component.
  • To characterize the electrochemical behavior of gel-immobilized Dd NiR.
  • To determine the substrate dependence of the enzyme's amperometric response to nitrite.

Main Methods:

  • Isolation of c-type hexaheme nitrite reductase from Desulfovibrio desulfuricans.
  • Immobilization of the enzyme onto a glassy carbon electrode.

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  • Electrochemical characterization, including cyclic voltammetry and amperometry.
  • Analysis of enzyme kinetics using Michaelis-Menten models.

    • Main Results:

    • Gel-immobilized Dd NiR exhibited enzymatic activity and amperometric response to nitrite.
    • The catalytic current density was dependent on nitrite concentration.
    • The enzyme's response followed Michaelis-Menten-type substrate dependence.
    • The NiR-electrode demonstrated potential for analytical nitrite determination.

    Conclusions:

    • Desulfovibrio desulfuricans NiR is a suitable candidate for biosensor development due to its substrate specificity, availability, and stability.
    • The NiR-electrode can be used for the analytical determination of nitrite in complex media.
    • Electrochemical immobilization of NiR provides a viable platform for biosensing applications.