ENZYME INHIBITION IN MICROFLUIDICS FOR RE-ENGINEERING BACTERIAL SYNTHESIS PATHWAYS
LARIOS BERLIN, DEAN EDWARD
RUBLOFF, GARY W
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Enzyme-functionalized biological microfluidic (EF-BioMEMS) systems are an emerging class of lab-on-chip devices that manipulate enzymatic pathways by localizing reaction sites in a microfluidic network. An EF-BioMEM system was fabricated to demonstrate biochemical enzyme inhibition. Further, design optimizations to the EF-BioMEM system have been proposed which improve system sensitivity and performance. The <italic>pfs</italic> enzyme is part of the quorum-sensing pathway that ultimately produces the bacterial signaling molecule AI-2. An EF-BioMEM system was fabricated to investigate the <italic>pfs</italic> conversion activity in the presence of a transition state analogue inhibitor. A reduction in enzyme conversion was measured in microfluidics for increasing inhibitor concentration that was comparable to the response expected on a larger scale. This EF-BioMEMS testbed is capable of investigating other compounds that inhibit quorum sensing. Design improvements were demonstrates that improve overall system responsiveness by minimizing unintended reactions from non-specifically bound enzyme. EF-BioMEMS signal-to-background performance increased from 0.72 to 2.43.