Figure 7.

Disubstituted luciferins are orthogonal to robust light-emitting luciferins, including (A) D-luc, (B) 4′-MeLuc, and (C) 7′-MeLuc. Mutants F247Y/S347G and 53 selectively processes the disubstituted analogs, while Fluc and mutant 87 prefer other luciferins. Bacterial cells expressing (A)-(B) mutant F247Y/S347G or Fluc or (C) mutant 53 or 87 were lysed and plated over a 96-well plate. Comparative emission values are plotted as the ratio of one luciferase over the other, on a log scale, for each substrate. Error bars represent the standard error of the mean for n ≥ 3 experiments. For each replicate, a single lysate was split among the wells and treated with each analog.