CAREER: Molecular and Genetic Characterization of the Early Steps of Toluene Utilization in Thauera sp. T1

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9733210 Coschigano The aromatic hydrocarbon toluene is a hazardous substance that poses health risks to humans, particularly to the central nervous system. Individuals with long term exposures may exhibit memory loss, attention and concentration deficiencies, and poor abstracting abilities. Toluene is one of the most water soluable components of gasoline and other fuels and has been listed by the U.S. Environmental Protection Agency as a priority pollutant. It is also a widely used industrial solvent, being used in a large number of chemical processes. This project will be the first use a combined genetic and molecular approach to study the non-oxygen mediated metabolism of an aromatic hydrocarbon using a bacterial isolate tentatively identified as a Thauera sp. that can degrade toluene under denitrifying conditions. Preliminary genetic and molecular studies have already proven useful in gaining insight into the mechanism of the toluene metabolic enzymes of strain T1, specifically the TutD gene product. This project will elucidate function of TutD and its regulation and identify additional enzymes in the pathway of toluene degradation using molecular genetic approaches. An essential component of this research is to provide an opportunity for both undergraduate student exposure to scientific research and for graduate student training. Ohio University has a number of programs to encourage undergraduates to participate in laboratory research and this proposal will help to provide an opportunity for such research. Opportunities for graduate student research at the Masters and doctoral level will also be provided.