<p>Our solution is to engineer bacteria containing specific enzymes known for defluorination and test that bacteria in water containing PFAS to see how well they can degrade this compound. Comparing our engineered bacteria with a control will allow us to effectively understand how the enzymes degrade the chemical and how well they break it up. The two enzymes we are testing are Fluoroacetate Dehalogenase and Haloacid Dehalogenase which will be inserted into E.Coli bacteria. Our end goal is to test the concentration of hydrogen ions with pH testing during experimentation to understand the effectiveness of degradation.</p>
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<p>In the future, we hope to test more enzymes such as P450, histine-ligated heme enzymes like TyrH and other hydroxylases or dehydrogenases to further our research. All enzymes interact with PFAS differently. Because of this, we hope to also see what byproducts are produced as a result of the degradation and use this information to see if certain enzymes contribute more to toxicity by creating toxic byproducts, or if there are safer enzymes that can partially degrade PFAS while also producing safe byproducts. Additionally, we want to enginner our bacteria in Pseudomonas in addition to E.Coli to test which type and strains of bacteria have a higher survival rate when degrading PFAS.</p>
<p>In the future, we hope to test more enzymes such as P450, histine-ligated heme enzymes like TyrH and other hydroxylases or dehydrogenases to further our research. All enzymes interact with PFAS differently. Because of this, we hope to also see what byproducts are produced as a result of the degradation and use this information to see if certain enzymes contribute more to toxicity by creating toxic byproducts, or if there are safer enzymes that can partially degrade PFAS while also producing safe byproducts. Additionally, we want to enginner our bacteria in Pseudomonas in addition to E.Coli to test which type and strains of bacteria have a higher survival rate when degrading PFAS. Adding in a fluorescent tag to our insert will tell us if the protein is being produced. This guarantees a greater understanding of whether or not the promoter is working and helps us understand the expression. To know if the protein is folded correctly and is functional, we can recover some of the protein out and conduct a western blot. This will allow us to see if the protein of the expected size is actually there which is more indicative of functionality. We could also try to conduct Fast Protein Liquid Chromatography (FPLC) by pulling out a lot of protein and doing a nocell reaction. To hopefully make this process more functiona, quick, and cheap, we hope to conduct further tests in the lab. </p>
