Dissolved Inorganic Carbon Uptake in <i>Thiomicrospira crunogena</i> XCL–2 is ATP–sensitive and Enhances RubisCO–mediated Carbon Fixation

............................................................................................................................. iv Chapter 1Introduction ........................................................................................................1 1.1 Hydrothermal Vent Habitat ................................................................................1 1.2 Thiomicrospira crunogena XCL-2 ....................................................................2 1.3 Ribulose-1,5-bisphosphate Carboxylase Oxygenase (RubisCO) ..........................................................................................................3 1.4 Carbonic Anhydrase...........................................................................................6 1.5 Carboxysomes ....................................................................................................6 1.6 Carbon Concentrating Mechanisms ...................................................................7 Chapter 2Materials and Methods .....................................................................................12 2.1 Reagents ...........................................................................................................12 2.2 Carboxysomal RubisCO Purification ..............................................................12 2.3 Silicone Oil Centrifugation: Measuring DIC, Fixed Carbon, , and pH ......15 2.4 ATP Quantification ..........................................................................................20 2.5 NADH Dehydrogenase (NDH-1) and Bc1 Complex Assays ..........................20 2.6 Statistical Analysis ...........................................................................................22 2.7 Phylogenetic Analysis ......................................................................................22 Chapter 3Results ..............................................................................................................23 3.1 Carboxysomal RubisCO Michaelis-Menten Kinetics ......................................23 3.2 Effect of Metabolic Inhibitors on DIC Uptake and Fixation ...........................24 Chapter 4Discussion ........................................................................................................29 4.1 References Cited ..............................................................................................34