A 28-year-old biochemistry research student is investigating the kinetic properties of a newly isolated enzyme from a bacterial culture. She measures the reaction velocity at various substrate concentrations and plots the data using a Lineweaver-Burk double reciprocal plot. The plot shows a y-intercept of 0.5 µmol⁻¹·min and an x-intercept of −2 mM⁻¹. Which investigation would be most appropriate to confirm the Km and Vmax values derived from this graphical analysis?