Bygd, Madison2022-09-132022-09-132022-05https://hdl.handle.net/11299/241569University of Minnesota M.S. thesis. 2022. Major: Microbial Engineering. Advisor: Lawrence Wackett. 1 computer file (PDF); 115 pages.Polyfluorinated compounds have become a popular topic in recent years for their widespread use and prominence as a pollutant in our environment. Their use in thousands of commercial products and chemicals has raised concern due to their resistance to degradation, coining them the “forever chemicals.” To better understand how these compounds are degraded and find microorganisms capable of remediation, biological degradative mechanisms were investigated. It was shown that 2,2-difluoro-1,3-benzodioxole, a common fluorinated moiety found in agricultural chemicals and pharmaceuticals, can be degraded and defluorinated by Pseudomonas putida F1. To better detect and screen for additional microbial related defluorination mechanisms, a color screen using alizarin and lanthanum was developed for use in biological contexts. Compatible with various media and solvents, this assay uncovered an additional 23 defluorination reactions initiated by P. putida F1. Further screening was continued with the E. coli ASKA library in which an additional novel defluorination reaction was found to be catalyzed by the enzyme PgpB. The color screen proved to be a highly effective and efficient method for detecting fluoride release by biological means in a high-throughput manner. The data presented in this thesis show many previously undocumented defluorination reactions and outline an efficient and effective method for detecting microbial based bioremediation of polyfluorinated chemicals.enbioremediationfluoride detectionfluorinated degradationmicrowell assayPFCPolyfluorinated ChemicalsThesis or Dissertation