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.

Luminescent Analysis of ATP: Modern Objects and Processes for Sensing

“…Potassium humate [61] Decrease of bioluminescence at concentration 0.5 g/L Isoflavonoids [62] High level of luciferase inhibition Divalent ions of various metals [63] Luminescence decrease in presence of Hg 2+…”

“…The effect of divalent ions on the bioluminescence of luciferase of fireflies has been established and it has been shown that an increase in the concentration of Mn 2+ , Ca 2+ or Mg 2+ ions does not affect the quantum yield and color of radiation, while the presence of Zn 2+ , Cd 2+ , Fe 2+ , Ni 2+ and Co 2+ ions cause a bathochromic shift, and luminescence decreased sharply in the presence of Hg 2+ [63].…”

Quantitative Analysis of Bioluminescence Optical Signal

“…Ten luciferins have been identified in bioluminescent organisms, along with several corresponding luciferases [3]. Firefly D-luciferin, coelenterazine, Cypridina luciferin, and their derivatives have been commercialized, and are primarily used in reporter assays, in vivo and in vitro imaging, and immunoassay [4][5][6][7][8][9]. Several companies, such as Promega and Toyobo, have also commercialized luciferase genes.…”

Glow‐type conversion and characterization of a minimal luciferase via mutational analyses

“…The brightness, thermostability, and small size of reporter proteins make them useful in applications. Their small size can reduce steric hindrance and misfolding of reporter‐fused proteins [3,4], making them amenable to genome insertion [5] and drug screening [6]. Additionally, the use of small luciferases can yield higher signals in bioluminescence resonance energy transfer (BRET)‐based assays [7].…”