ENZYME INHIBITION IN MICROFLUIDICS FOR RE-ENGINEERING BACTERIAL SYNTHESIS PATHWAYS

Abstract

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.