Bacterial P450-catalyzed polyketide hydroxylation on a microfluidic platform

Biotechnol Bioeng. 2004 Nov 20;88(4):528-35. doi: 10.1002/bit.20285.


The incorporation of a multicomponent, cofactor-dependant P450 into a microfluidic biochip is demonstrated. The PikC hydroxylase Streptomyces venezuelae was incorporated into a PDMS-based microfluidic channel. The enzyme was immobilized to Ni-NTA agarose beads via in situ attachment following the addition of the beads to the microchannel. The enzyme loading was approximately 6 microg per mg of beads resulting in a microchannel loading of 10.7 mg/mL. This high enzyme loading enabled the rapid hydroxylation of the macrolide YC-17 to methymycin and neomethymycin in about equal amounts with a conversion of >90% at a flow rate of 70 nL/min. This high reactivity allowed rapid hydroxylation reactions to be performed with short residence times, which is critical for complex enzymes with limited inherent stability.

Publication types

  • Evaluation Study
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bioreactors
  • Catalysis
  • Cytochrome P-450 Enzyme System / chemistry*
  • Enzyme Activation
  • Enzymes, Immobilized
  • Equipment Design
  • Equipment Failure Analysis
  • Hydroxylation
  • Macrolides / chemical synthesis*
  • Macrolides / chemistry*
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods*
  • Streptomyces / enzymology*


  • Enzymes, Immobilized
  • Macrolides
  • YC-17 macrolide
  • methymycin
  • neomethymycin
  • Cytochrome P-450 Enzyme System