Palette of Luciferases: Natural Biotools for New Applications in Biomedicine

Abstract

Optoanalytical methods based on using genetically encoded bioluminescent enzymes,luciferases, allow one to obtain highly sensitive signals, are non-invasive, and require no external irradiation. Bioluminescence is based on the chemical reaction of oxidation of a low-molecular-weight substrate (luciferin) by atmospheric oxygen, which is catalyzed by an enzyme (luciferase). Relaxation of the luciferin oxidation product from its excited state is accompanied by a release of a quantum of light, which can be detected as an analytical signal.The ability to express luciferase genes in various heterological systems and high quantum yields of luminescence reactions have made these tools rather popular in biology and medicine. Amongseveral naturally available luciferases, a few have been found to be useful for practicalapplication. Luciferase size, the wavelength of its luminescence maximum, enzyme thermostability, optimal pH of the reaction, and the need for cofactors areparameters that may differ for luciferases from different groups of organisms, and this fact directly affects the choice of the application area for each enzyme. It is quite important to overview the whole range of currently available luciferases based ontheir biochemical properties before choosing one bioluminescent probe suitable for a specific application.

Luciferins Under Construction: A Review of Known Biosynthetic Pathways

“…Beetle bioluminescence systems belong to one of the most practically important and well-understood groups of bioluminescence reactions. The ability to emit light has evolved in several Coleoptera families including fireflies (Lampyridae), click beetles (Elateridae) and railroad worms (Phengodidae), with the light emission wavelength ranging from green to red (540-640 nm) depending on the beetle species (Viviani et al, 2011;Kotlobay et al, 2020). All known beetle bioluminescence system depend on a common substrate (S)-2-(6-hydroxy-2-benzothiazolyl)-2-thiazoline-4carboxylic acid, commonly known as D-luciferin (Figure 1), which consists of two structural units, benzothiazole and thiazoline rings (Shimomura and Yampolsky, 2019).…”

“…A number of coelenterazine-utilizing luciferases are well known, including those of soft corals (Renilla), copepods (Gaussia, Metridia) and decapods (Oplophorus) (Kotlobay et al, 2020), while vargulin-dependent luciferases have been derived only from an ostracod lineage Cypridinidae (Hensley et al, 2021). Coelenterazine also serves as the light emitter in the Ca 2+binding photoproteins, such as aequorin and obelin (Head et al, 2000).…”

“…However, two recent publications have revealed a surprising similarity between luciferases of phylogenetically distant organisms: the luminous brittle star Amphiura filiformis (Echinodermata), tunicates Pyrosoma atlanticum (Chordata) and the sea pansy Renilla (Cnidaria), indicating that all three luciferases may have evolved convergently from homologous dehalogenases (Loening et al, 2006;Delroisse et al, 2017;Tessler et al, 2020). Bioluminescence maxima of all naturally occurring imidazopyrazinone-dependent enzymes lie in the blue region of the visible spectrum (450-490 nm), although in some cases the color of luminescence is biochemically altered by an energy transfer to fluorescent protein that interacts with the luciferase (Kotlobay et al, 2020).…”

Miniaturization of Bright Light-Emitting Luciferase ALuc: picALuc

“…It has been previously reported that the luminescence intensity of GLuc is 200-fold higher than those of FLuc and RLuc, and its thermostability is remarkably high. 10,11 More recently, small luciferases emitting bright light, such as TurboLuc, 12 Nano-Luc, 13 and artificial luciferase (ALuc) 14,15 were developed. TurboLuc 12 is a modified luciferase derived from the copepod Metridia pacifica (MpLuc1), which has a sequence similar to that of GLuc.…”

“…14,15 The small luciferases, such as GLuc and NanoLuc, have been recently utilized because their small sizes can reduce the steric hindrance of the luciferase-fused proteins, leading to the increased maintenance of the native functions of the proteins. 11,17 Additionally, their small sizes have an advantage in the bioluminescence resonance energy transfer (BRET)based assays because the BRET signal is inversely proportional to the sixth power of the distance between luciferase and fluorescent chromophore. 18 The bright light emission can lead to extremely sensitive detections; moreover, their high thermostabilities are valuable for practical applications such as in public health inspections, food safety and diagnosis, and transportation.…”