High levels of toxic contamination of perfluorinated compounds, PFCs, found in the Southern Colorado Springs Metro Area have been measured in drinking water and degraded in organisms found originating at Peterson Air Force Base, one of approximately 2,000 Department of Defense known chemical spill sites. They have been known to cause significant health effects such as affecting fetal growth and development, cancer, and injury to vital organs in exposed populations. PFCs are both long-lived and toxic, and limited governmental resources for impact investigation make the proposed work both urgent and of inherently high impact to the citizenry of southern Colorado. Methods such as filtration and carbon sorption are ineffective and expensive, so alternative methods are needed to remove them. Here we propose a method of bioremediation to defluorinate PFCs using enzymes bioengineered to degrade per-fluorinated compounds. This method is a better alternative than the previously proposed methods since the enzyme works to remove the fluorine atoms, as fluoride ions, from the compound. This makes degradation more efficient and better for the environment rather than re-locating the toxic compound to another site, which would thus increase the total amount of PFC contamination. A haloalkane dehalogenase protein from a marine Rhodobacteracea was expressed using standard bacteriological protein expression. The enzyme was then purified using affinity chromatography, purity was monitored using FPLC and SDS-Page gels. We are currently modifying known dehalogenation assays to measure the inherent de-fluorination of our enzyme before testing optimized enzymes. Our next steps begin by modeling the active site to drive the bioengineering of the enzyme by locating and identifying important amino acids for mutagenesis.

Bioengineering of a haloalkane dehalogenase for the bioremediation of perfluorinated compounds

ACS Spring 2022

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