The Michaelis–Menten Equation and Its Linear Transformations Revisited

ABSTRACT Twelve published enzyme kinetic datasets were fitted with the Michaelis–Menten equation via non‐linear regression and with linear transformations (Lineweaver–Burk, Eadie–Hofstee, and Hanes–Woolf) via linear regression to compare parameter estimates ( V max and K M ). Model performances were compared based on the sum of squared error values calculated on the untransformed scale. The best method to obtain V max and K M was to fit the Michaelis–Menten equation via nonlinear regression. Among linear transformations, the Hanes–Woolf yielding the closest estimates to the non‐linear fit in seven out of 12 datasets, and the Eadie–Hofstee in five out of 12. Lineweaver–Burk, despite its widespread use, consistently performed the poorest, providing no closest estimates in any dataset. We concluded that the Lineweaver–Burk transformation may be utilized for data visualization; however, it is not recommended for estimating kinetic parameters, at least without weighted linear regression. If simple linear regression is to be employed for the determination of V max and K M , Hanes–Woolf transformation presents a preferable alternative, albeit with certain inherent limitations that must be considered. It can also be used for educational purposes (Excel solutions of this study are available at https://drive.google.com/drive/u/0/folders/1KfyvXTGG‐3Hgc9MVlXiKQH1YHbpAYwNm for further reference). Researchers should apply non‐linear regression to obtain V max and K M .