A statistical comparison of parameter estimation for the Michaelis–Menten kinetics of human placental hexosaminidase

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Abstract
To enable the most effective method of kinetic discrimination between a group of isozymes such as those of human placental hexosaminidases (HEX), three methods estimating the parameters of the Michaelis–Menten equation were evaluated. Computer-simulated experiments were performed under various conditions. They indicated that, in the presence of constant absolute or relative errors, the method of unweighted nonlinear least squares yielded slightly more precise and accurate parameters than the method of the direct linear plot. Parameters calculated from the Lineweaver–Burk plot were very imprecise and inaccurate. The direct linear plot was comparatively resistant to outlier observations; however, only when outliers were substantial did the method become superior to nonlinear least squares. The calculation of a confidence limit is necessary for the evaluation of any resulting differences in the kinetic parameters for a set of isozymes. This can easily be calculated from either the Lineweaver–Burk plot or the nonlinear least-squares method. However, those calculated from the Lineweaver–Burk plot are biased, especially at higher levels of experimental errors. Therefore, the nonlinear least-squares method is the one most suited for the discrimination of a group of enzymes based on their kinetic parameters.