High-level expression, purification, and characterization of Staphylococcus aureus dihydroorotase (PyrC) as a cleavable His-SUMO fusion

► The purification of the Staphylococcus aureus PyrC enzyme using a His-SUMO tag is described. ► Protein solubility and yield were high with the use of the His-SUMO tag. ► Improved tag cleavage was achieved with HRV 3C protease over SUMO protease. ► Forward and reverse kinetic rate constants for SaPyrC were calculated experimentally. ► Optimal pH values for forward and reverse reactions were determined.