Expression, purification and luminescence properties of coelenterazine-utilizing luciferases from Renilla, Oplophorus and Gaussia: Comparison of substrate specificity for C2-modified coelenterazines

► The coelenterazine-utilizing luciferases show different luminescence characteristics. ► The C2-modified coelenterazine analogues are used for the luciferase reaction. ► The emission peaks are not affected by the C2-substituted coelenterazines. ► Gaussia luciferase has the highest luminescence activity among luciferases. ► The catalytic domain of Oplophorus luciferase shows broad substrate specificity.