Zinaida M. Kaskova, Felipe A. Dörr, Valentin N. Petushkov, Konstantin V. Purtov, Aleksandra S. Tsarkova, Natalja S. Rodionova, Konstantin S. Mineev, Elena B. Guglya, Alexey Kotlobay, Nadezhda S. Baleeva, Mikhail S. Baranov, Alexander S. Arseniev, Josef I. Gitelson, Sergey Lukyanov, Yoshiki Suzuki, Shusei Kanie, Ernani Pinto, Paolo Di Mascio, Hans E. Waldenmaier, Tatiana A. Pereira, Rodrigo P. Carvalho, Anderson G. Oliveira, Yuichi Oba, Erick L. Bastos, Cassius V. Stevani, Ilia V. Yampolsky

Bioluminescent fungi are spread throughout the globe, but details on their mechanism of light emission are still scarce. Usually, the process involves three key components: an oxidizable luciferin substrate, a luciferase enzyme, and a light emitter, typically oxidized luciferin, and called oxyluciferin. We report the structure of fungal oxyluciferin, investigate the mechanism of fungal bioluminescence, and describe the use of simple synthetic α-pyrones as luciferins to produce multicolor enzymatic chemiluminescence. A high-energy endoperoxide is proposed as an intermediate of the oxidation of the native luciferin to the oxyluciferin, which is a pyruvic acid adduct of caffeic acid. Luciferase promiscuity allows the use of simple α-pyrones as chemiluminescent substrates.