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A direct electrode-driven P450 cycle for biocatalysis

V Reipa1, M P Mayhew, V L Vilker

  • 1Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.

Proceedings of the National Academy of Sciences of the United States of America
|February 12, 1998
PubMed
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This study presents a novel bioreactor for efficient cofactor regeneration in redox enzyme catalysis. It demonstrates sustained biocatalysis using electrochemistry to power cytochrome P450cam, achieving high product formation rates.

Area of Science:

  • Biocatalysis and Enzyme Engineering
  • Electrochemical Synthesis
  • Organic Chemistry

Background:

  • Redox enzymes offer significant potential for synthesizing high-value organic compounds.
  • Efficient cofactor regeneration is crucial for the economic viability of biocatalytic processes.
  • Current cofactor regeneration methods often face limitations in scalability and efficiency.

Purpose of the Study:

  • To develop an innovative bioreactor for direct cofactor regeneration in redox enzyme systems.
  • To investigate the direct electrochemical supply of reducing power to cytochrome P450cam (CYP101).
  • To achieve sustained and efficient biocatalysis using an integrated electrochemical-enzymatic system.

Main Methods:

  • A bioreactor was designed utilizing an antimony-doped tin oxide electrode for direct electron transfer to putidaredoxin, the redox partner of CYP101.

Related Experiment Videos

  • Oxygen, essential for the catalytic cycle, was generated electrochemically via water electrolysis at a platinum counter electrode.
  • The system employed purified cytochrome P450cam (P450cam) for the biocatalytic transformation.

    • Main Results:

    • A continuous catalytic cycle was successfully sustained for over 5 hours.
    • The system achieved more than 2,600 enzyme turnovers, demonstrating high operational stability.
    • A maximum product formation rate of 36 nmol of 5-exo-hydroxycamphor/nmol of CYP101 per minute was recorded.

    Conclusions:

    • Direct electrochemical regeneration of cofactors is a viable and efficient strategy for powering redox enzymes.
    • This integrated electrochemical-enzymatic approach offers a promising platform for sustainable and scalable biocatalysis.
    • The developed bioreactor system significantly enhances the efficiency and turnover of cytochrome P450cam-mediated reactions.