title: Energetics of proline racemase: racemization of unlabeled proline in the unsaturated, saturated, and oversaturated regimes.
abstract: The interconversion of L- and D-proline catalyzed by proline racemase has been studied. The entire time course of the approach to equilibrium has been followed. After a short time the product concentration is significant, and the reaction runs under reversible conditions. As the total substrate concentration is increased, the system moves from the unsaturated regime into the saturated regime. At very high substrate levels under the reversible conditions used, the rate constant for substrate racemization falls, as the system moves into the "oversaturated" regime. Here, the net rate of the enzyme-catalyzed reaction is limited by the rate of return of the free enzyme from the form that liberates product back to the form that binds substrate. The results are analyzed in terms of the simple mechanism (table; see text) and illustrate the additional information that is available from reactions studied under reversible conditions. In the unsaturated region the value of the second-order rate constant kU (equivalent to kcat/Km) is 9 X 10(5) M-1 s-1 in each direction. In the saturated region, kcat = kcat = 2600 s-1 and Km = 2.9 mM. In the oversaturated region, the rate constant kO is 81 M s-1. The substrate concentration at which unsaturated and saturated terms contribute equally is 2.9 mM, and the substrate concentration at which saturated and oversaturated terms contribute equally is 125 mM.
mesh_terms: D000591:Amino Acid Isomerases / Q000378:metabolism*; D026921:Amino Acid Transport Systems, Neutral; D003013:Clostridium / Q000201:enzymology*; D007700:Kinetics; D008433:Mathematics; D026901:Membrane Transport Proteins / Q000302:isolation & purification; D009904:Optical Rotation; D011485:Protein Binding; D013237:Stereoisomerism
chemical_list: D026921:Amino Acid Transport Systems, Neutral; D026901:Membrane Transport Proteins; C033082:proline transporter; D000591:Amino Acid Isomerases; C022451:proline racemase