Enzyme inhibition in microfluidics for re-engineering bacterial synthesis pathways |
Posted on:2010-11-13 | Degree:M.S | Type:Thesis |
University:University of Maryland, College Park | Candidate:Larios Berlin, Dean | Full Text:PDF |
GTID:2441390002973864 | Subject:Engineering |
Abstract/Summary: | |
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 pfs 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 pfs 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. |
Keywords/Search Tags: | Enzyme, System, Ef-biomems |
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