CAREER: Mechanistic Studies of the Alkanesulfonate Monooxygenase System

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Certain soil bacteria will utilize aliphatic sulfonates in the environment to obtain sulfate if standard sources are not available. The enzyme system involved in this process is comprised of a flavin-reductase (SsuE) involved in providing reduced flavin to the monooxygenase enzyme (SsuD) that is directly involved in the desulfonation reaction. This fascinating enzyme system belongs to an increasing number of two-component enzyme systems that utilize flavin as a substrate rather than a bound prosthetic group. These systems include proteins involved in the degradation of herbicides, the synthesis of antibiotics, desulfurization of pollutants, and bioluminescence. The broad objective of these studies is to elucidate the mechanism of the alkanesulfonate monooxygenase system. The primary goal of this research focuses on two highly interesting features of this enzyme system: 1) the mechanism of C-S bond cleavage by SsuD 2) the reduced flavin transfer mechanism between SsuE and SsuD. Relating the results obtained from these studies of the alkanesulfonate monooxygenase proteins to other flavin transfer systems will enhance our knowledge of a family of enzymes with highly diverse functions. The educational goal of this project is to enhance the Auburn Science in Motion program by introducing biochemical experiments to the existing curriculum. The Science in Motion program was established by the Alabama State Legislature to bring technologically advanced laboratory experiments to local high school classrooms. The educational component will be implemented through three distinct activities: 1. experimental development 2. graduate student outreach 3. direct training to high school educators. The impact of this project will affect students from all levels and introduce those students from underrepresented groups and economically disadvantaged school districts to innovative experiments, which can increase student interest in science and influence their future educational decisions.