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  • Title: Bacterial P450-catalyzed polyketide hydroxylation on a microfluidic platform.
    Author: Srinivasan A, Bach H, Sherman DH, Dordick JS.
    Journal: Biotechnol Bioeng; 2004 Nov 20; 88(4):528-35. PubMed ID: 15459906.
    Abstract:
    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.
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