The invention relates to a bioluminescent substrate suitably usable in a series of artificial luciferases (ALuc), and the invention provides a wavelength-shifted spectrum with a selective high intensity luminescence and high luminescence stability obtained by the use of the substrate together with ALuc. The luminescent substrate for ALuc obtained by the invention can be included together with a suitable luminescence solution in a luminescence kit. The bioluminescent substrate for ALuc of the invention can exhibit unprecedented excellent luminescence specificity and functionality in the conventional bioluminescence probe, two-hybrid assay, bioluminescent capsule, and reporter gene assay.