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

9733210 Coschigano芳香烃甲苯是一种对人类健康构成危险的物质，特别是对中枢神经系统。长期接触的人可能会表现出记忆力丧失、注意力和注意力不足以及抽象能力低下。甲苯是汽油和其他燃料中最易溶于水的组分之一，已被美国环境保护局列为优先污染物。它也是一种广泛使用的工业溶剂，用于大量的化学过程。这个项目将首次使用遗传和分子相结合的方法来研究芳香烃的非氧介导的代谢，使用的是一种暂时被鉴定为特乌埃拉细菌的细菌分离物。可以在反硝化条件下降解甲苯。初步的遗传和分子研究已被证明有助于深入了解菌株T1的甲苯代谢酶的机制，特别是TutD基因产物。本项目将阐明TutD的功能及其调控，并利用分子遗传学方法确定甲苯降解过程中的其他酶。这项研究的一个重要组成部分是为本科生接触科学研究和研究生培训提供机会。俄亥俄大学有许多鼓励本科生参与实验室研究的计划，这项提议将有助于为此类研究提供机会。还将提供硕士和博士级别的研究生研究机会。

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.