Kinetic Analyses of Enzyme Reaction Curves with New Integrated Rate Equations and Applications

نویسندگان

  • Xiaolan Yang
  • Gaobo Long
  • Hua Zhao
  • Fei Liao
چکیده

A reaction system of Michaelis-Menten enzyme on single substrate can be characterized by the initial substrate concentration before enzyme action (S0), the maximal reaction rate (Vm) and Michaelis-Menten constant (Km), besides some other required parameters. The estimations of S0, Vm and Km can be used to measure enzyme substrates, enzyme activities, epitope or hapten (enzyme-immunoassay), irreversible inhibitors and so on. During enzyme reaction, the changes of substrate or product concentrations can be monitored; continuous monitor of such changes provides a reaction curve while discontinuous monitor of such changes provides signals just for the starting point and the terminating point of enzyme reaction. It is an end-point method when only signals for the starting point and the terminating point are analyzed. It is a kinetic method when a range of data from a reaction curve are analyzed, and can be classifieid into the initial rate method and kinetic analysis of reaction curve. The initial rate method only analyzes data for initial rate reaction whose instantaneous rates are constants; kinetic analysis of reaction curve analyzes data whose instantaneous rates show obvious deviations from the initial rate (Bergmeyer, 1983; Guilbault, 1976; Marangoni, 2003). To estimate those parameters of an enzyme reaction system, kinetic analysis of reaction curve is favoured because the analysis of one reaction curve can concomitantly provide Vm, S0 and Km. Hence, methods for kinetic analysis of reaction curve to estimate parameters of enzyme reaction systems are widely studied.

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تاریخ انتشار 2012