Figure 8: The kinetic parameters Km and Vmax were calculated by nonlinear regression analysis of experimental steady-state data using the computer program GraFit, Erithacus Software. Plot of the reaction velocities (V) vs. substrate concentration (S: 0.00–2.5 mM) fitted to the Michaelis– Menten equation (R square=0.99) and determination of Km and Vmax of purified L-asparaginase for L-asparagine by non-linear regression analysis of experimental steady-state data.The corresponding Lineweaver–Burk plot for L-asparaginase catalyzed reaction.