Abstract 1839 Expression of F420 dependent glucose-6-phosphate dehydrogenase from Nocardioidaceae bacterium

F420 dependent glucose-6-phosphate dehydrogenase (FGD), although originally discovered in Nocardia, has been extensively studied in Mycobacteria due to the health relevance for tuberculosis (TB) treatment. FGD catalyzes the conversion of glucose-6-phosphate (G6P) to 6-phosphogluconolactone, using oxidized F420 cofactor, which becomes reduced during catalysis. The focus of this project is on FGD from Nocardioidaceae bacterium (FGD-Noca), which shows activity towards additional sugar-6-phosphates, such as glucose-6-phosphate (G6P), D-mannose-6-phosphate (M6P), and D-fructose-6-phosphate (F6P), unlike the mycobacterial enzyme which only displays specificity towards G6P. The major goal of our research is to characterize the FSDs using steady-state and pre steady-state kinetic methods to provide detailed mechanistic insights for the FSDs. Our initial goal of this project is to express and purify FGD-Noca in order to kinetically characterize the hydride transfer reaction mechanism of wtFGD-Noca. The proposed experiments and preliminary results will be described here. Our project is currently funded by WELCH foundation.