Topic Area: 4e. PFAS Source and Forensic Considerations

Formatted Title:

Discovery of Novel Defluorinases for PFAS Bioremediation Using Ultrahigh-Throughput Droplet Microfluidics

Background/Objectives:

Per- and polyfluoroalkyl substances (PFAS) are a very large and diverse class of recalcitrant compounds (also known as forever chemicals) which are essentially of anthropic origin. Industrially produced PFAS are used for a wide range of applications in all areas of human activity, and now represent ubiquitous contaminants in the environment. A main feature of PFAS is their extensive persistence conferred by their halogenation; the carbon-fluorine bond being the most stable covalent single bond known. Today, these compounds have been associated with serious adverse effects on human health, and there is a growing need to decontaminate the environment of these pollutants. Our goal is to discover PFAS-degrading enzymes either through environmental sampling or via directed evolution.

Approach/Activities:

Complementarily to current physical and chemical remediation processes, enzymatic degradation represents an attractive approach for PFAS remediation. However, enzymatic degradation of the C–F bonds of such compounds hasn’t been fully assessed. Naturally, the ability for a biological system (e.g., bacteria) to break down “foreign” environmental pollutants relies on the inherent contact with such a substance. This proves to be rather challenging when dealing with synthetic pollutants like PFAS.

Biodegradation of fluorinated compounds entails the fact that the molecules in question must break down the carbon-fluorine bonds in the various PFAS forms, releasing fluoride ions. Therefore, monitoring fluoride production could allow monitoring of enzyme activity. In this view, our team developed "FluorMango" biosensor, a fluorogenic structure switching aptamer specifically emitting fluorescence in the presence of fluoride and is compatible with droplet-based microfluidic ultrahigh-throughput screening platforms. This allows for rapid screening of novel defluorinases that could be adapted to downstream characterization and industrial applications.

Enrico Bocconetti

Affiliation

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(Group 2, Poster Board #6) Discovery of Novel Defluorinases for PFAS Bioremediation Using Ultrahigh-Throughput Droplet Microfluidics