A bioluminogenic assay system including: providing a bioluminogenic substrate incorporating a beta-lactam antibiotic, a bioluminescence initiating compound, and a chemical linkage joining the beta-lactam antibiotic to the bioluminescence initiating compound; exposing the bioluminogenic substrate to a beta-lactamase enzyme that catalyzes the release of the bioluminescence initiating compound from the bioluminogenic substrate; co-exposing the bioluminogenic substrate to a bioluminescence indicator reaction that employs the bioluminescence initiating compound as a substrate; and detecting a light from the bioluminescence indicator reaction as a measure of the activity of the beta-lactamase enzyme.