Mechanically Assisted Bioluminescence with Natural Luciferase.

Mechanochemical analogues have been recently established for several enzymatic reactions, however they require periodic interruption of the reaction for sampling, dissolution, and (bio)chemical analysis to monitor their progress. By applying a mechanochemical protocol to induce bioluminescence analogous to that used by the marine ostracod Cypridina ( Vargula ) hilgendorfii here we demonstrate that the light emitted by a bioluminescent reaction can be used for direct monitoring of the progress of a mechanoenzymatic reaction without sampling. Mechanical treatment of Cypridina luciferase and luciferin generates bright blue light which can be readily spectroscopically detected and analyzed. We establish that this mechanically assisted bioluminescence proceeds through a mechanism identical to that of bioluminescence in solution however it has higher activation energy due to being diffusion-controlled in the viscous matrix. The results open prospects for application of luciferases as light-emissive reporters for mechanoenzymatic reactions, either as chemically coupled enzymatic labels to the reactant or as an internal standard added in the reaction mixture.