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Steroid bioconversion in a microemulsion system.

A J Smolders1, H M Pinheiro, P Noronha

  • 1Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, Lisboa, Portugal.

Biotechnology and Bioengineering
|December 5, 1991
PubMed
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This study optimized steroid dehydrogenation using Arthrobacter simplex cells in a microemulsion system. Thawed cells achieved 98% conversion, demonstrating efficient biocatalysis for steroid transformation.

Area of Science:

  • Biocatalysis and Enzyme Engineering
  • Microemulsion Systems for Biotransformations
  • Steroid Chemistry and Metabolism

Background:

  • High concentrations of steroids pose challenges for enzymatic transformations.
  • Microemulsion systems offer unique environments for solubilizing hydrophobic substrates.
  • Arthrobacter simplex is a known biocatalyst for steroid dehydrogenation.

Purpose of the Study:

  • To investigate the Delta(1,2)-dehydrogenation of high concentrations of 16-methyl-Reichstein's compound S-21-acetate (16MRSA).
  • To optimize the microemulsion system for efficient steroid biotransformation using Arthrobacter simplex.
  • To characterize the kinetic parameters and operational stability of the biocatalytic system.

Main Methods:

  • Utilized heat-dried and thawed Arthrobacter simplex cells as biocatalysts.

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  • Employed a microemulsion system composed of an organic phase (benzene derivatives), aqueous phase (cells), and phospholipid surfactant.
  • Included menadione as an external electron acceptor and studied reaction kinetics (Michaelis-Menten) and operational stability.

    • Main Results:

    • Achieved high conversion rates (98%) within 14-16 hours using thawed cells at optimal concentrations.
    • Determined Michaelis-Menten kinetic parameters for 16MRSA (V'(max)=2.06 g/g dry weight h, K'(m)=18.9 g/L) and menadione (V'(max)=4.97 g/g dry weight h, K'(m)=1.91 g/L).
    • Observed strong product inhibition and dependent optimal menadione concentration on steroid concentration; activity decreased over 48 hours in a stirred tank reactor.

    Conclusions:

    • Thawed Arthrobacter simplex cells in a microemulsion system are highly effective for Delta(1,2)-dehydrogenation of concentrated 16MRSA.
    • The microemulsion system facilitates high substrate loading and efficient biocatalysis.
    • Operational stability in a stirred tank reactor is influenced by stirring rate, suggesting potential for process optimization.