27] Applications of Renilla bioluminescence: An introduction

Abstract

This chapter focuses on the application of Renilla bioluminescence system including the characteristics of the proteins and substrates involved in the light-emitting process. The molecular basis for Renilla bioluminescence, as well as certain features of its nerve-linked control are also described. The requirement for 3´,5´-diphosphoadenosine (PAP) in Renilla bioluminescence involved a cofactor in the conversion of luciferyl sulfate to coelenterate-type luciferin by the enzyme luciferin sulfokinase. The assay for PAP involves a coupled-enzyme luminescence assay utilizing luciferyl sulfate as the substrate. Renilla luciferase is purified to homogeneity from crude extracts with an overall recovery of 24%. Luciferase also contains three free SH groups but no disulfide linkages. Renilla luciferase can be classified as an oxygenase (O 2 ) as it is apparently incorporated into one of the products, oxyluciferin, while a second product, carbon dioxide (CO 2 ) is derived from carbon 3 of luciferin. The initial product of the reaction is an electronic excited state of luciferaseoxyluciferin-monoanion complex that relaxes to its ground state with the production of blue light.