Revisiting linearization in enzyme kinetics: how experimental design determines parameter accuracy

This preprint investigates how experimental design influences parameter estimation in linearized enzyme kinetics. Using systematic numerical simulations, we compare the Lineweaver–Burk, Hanes–Woolf, and Eadie–Hofstee transformations under a wide range of substrate distributions, noise levels, and sampling strategies. The results show that estimation accuracy depends primarily on experimental design rather than on the linearization method itself, providing practical guidance for improving the reliability of kinetic parameter estimation.