Cloning and characterization of luciferase-like enzyme of non-bioluminescent Coleoptera and their relationships with the origin of luminescent activity

Abstract

Bioluminescence, the production of light by several living organisms, is a biochemical reaction in which the chemical energy is converted in light. In this reaction the substrate (luciferin) is oxygenated via an enzyme, luciferase, generating energy-rich peroxidic intermediates, whose spontaneous decomposition generates singlet electronically excited products which decay emitting a photon of visible light. Among terrestrial organisms the bioluminescence is more abundant in insects especially in beetles. In this group there is a wide range of pattern and color that has communicative purposes intra and inter specific, however, the bioluminescence in different families shares a chemical reaction that uses the same benzothiazolic luciferin, ATP and homologous luciferases.The bioluminescence has been used, for a long time, as tool for investigating and understanding numerous questions ranging from ecology, evolution, gene expression and enzymology. In the last decades it became a sensible and versatile tool in direct applications involving measurement of ATP for analytical and clinical purposes besides a new set of applications using the firefly luciferase gene as one of the most sensitive reporters of gene expression in living cells and tissues. In respect to the origin and molecular evolution of luciferases, the current view suggests that they arose from an AMP-CoA ligase family. However, there is no consistent connection between non-luminescent ligases and the luciferases, so the knowledge of enzyme structure is an interesting subject to infer how the luminescent function of luciferases originated. This project has the aim to clone and characterize luciferase enzyme-like (AMP-ligases) of non-bioluminescent beetle. These results will be essential to understand the key of bioluminescence, trace an evolutive way of bioluminescence that can be useful in the development and improvement of new luciferases.