%0 PDF %T Single Molecule Enzymology Using Femtoliter Arrays. %A Mogalisetti, Pratyusha. %8 2017-04-20 %R http://localhost/files/6m312169c %X Abstract: This thesis demonstrates the application of optical fiber based femtoliter arrays for single molecule enzymology studies. Study of enzymes at single molecule level provides us with mechanistic insights that cannot be obtained using traditional bulk studies. Several approaches for isolating and studying single enzyme molecules have been reported in the literature. These methods are detailed in Chapter 1 with an emphasis on femtoliter array platforms for single molecule isolation. Chapter 2 describes the application of optical fiber based arrays to study the stochastic binding and release of the inhibitors to single β-galactosidase molecules. The competition between the substrate and inhibitor for the enzyme active site is studied at single molecule level. The effect of inhibition on enzyme activity is also described. In Chapter 3, the α-complementation reaction of β-galactosidase was studied at single molecule resolution. In a novel application of the femtoliter arrays, the numbers of active molecules formed in the complementation reaction were directly counted and the stoichiometry of complementation reaction was determined. Chapter 4 discusses the effect of temperature on single molecules of wild type and single surface cysteine variants of β-glucuronidase. The identification of the thermal switching property of a fraction of the enzyme population in some variants is reported. Finally, chapter 5 discusses the current progress towards identifying hetero-oligomers with unnatural backbones that can catalyze notoriously difficult reactions. The approach of screening large oligomer libraries for the presence of rare catalysts using the optical fiber femto-well array platform is also described.; Thesis (Ph.D.)--Tufts University, 2015.; Submitted to the Dept. of Chemistry.; Advisor: David Walt.; Committee: Krishna Kumar, Joshua Kritzer, and Jeff Gelles.; Keywords: Chemistry, and Biochemistry. %[ 2022-10-11 %9 Text %~ Tufts Digital Library %W Institution