Luciferase
[CAS# 9014-00-0]

Identification

• Classification: Biological >> Proteins and peptides >> Enzyme and activity assay
• Name: Luciferase
• Synonyms: 1: PN: CN103239735 FIGURE: 4A claimed sequence; Alkanal monooxygenase (FMN-linked); Bacteria luciferase; Bacterial luciferase; E.C. 1.14.14.3; Luciferase (Photobacterium leiognathi); Photorhabdus luminescens luciferase; Vibrio fischeri luciferase; Vibrio harveyi luciferase
• CAS Registry Number: 9014-00-0
• EC Number: 232-751-7

Safety Data

• Hazard Symbols: GHS08 Danger
• Hazard Statements: H334
• Precautionary Statements: P261-P284-P501

Discovory and Applicatios

Luciferase is a class of enzymes responsible for bioluminescence, the natural emission of light by living organisms. The phenomenon of bioluminescence attracted scientific attention in the nineteenth century, when researchers sought to understand how organisms such as fireflies, luminous bacteria, and certain marine animals produce visible light at ambient temperatures. Early systematic studies established that light emission was not due to heat but to a chemical reaction. By the late nineteenth and early twentieth centuries, experiments demonstrated that bioluminescence required both a heat-stable small molecule substrate, later termed luciferin, and a heat-labile protein component, which came to be known as luciferase. The term luciferase was introduced to describe the enzyme that catalyzes the oxidation of luciferin, leading to light emission.

The most influential early work on luciferase was carried out using fireflies, particularly species such as Photinus pyralis. Researchers showed that firefly luciferase catalyzes the ATP-dependent oxidation of D-luciferin in the presence of molecular oxygen and magnesium ions, producing oxyluciferin, carbon dioxide, adenosine monophosphate, and visible light. This discovery established luciferase as a distinct enzyme with defined substrate specificity and cofactor requirements. Parallel studies on marine organisms and luminous bacteria revealed that luciferases from different species vary in structure and reaction mechanism, even though they all produce light. Bacterial luciferase, for example, catalyzes the oxidation of reduced flavin mononucleotide and a long-chain aldehyde, illustrating that the term luciferase encompasses a family of evolutionarily diverse enzymes rather than a single protein.

As biochemical techniques advanced in the mid-twentieth century, luciferase was purified and characterized in greater detail. The determination of amino acid sequences and three-dimensional structures, particularly for firefly luciferase, provided insight into enzyme catalysis and structure-function relationships. These studies confirmed that luciferase is a soluble cytosolic enzyme and clarified the molecular basis of its high quantum efficiency, meaning that a large proportion of chemical energy is converted into visible light. The cloning of luciferase genes in the late twentieth century marked a major turning point, allowing recombinant expression of luciferase in heterologous systems.

The applications of luciferase expanded dramatically following the development of molecular biology and genetic engineering. Luciferase genes became widely used as reporter genes to monitor gene expression, promoter activity, and signal transduction pathways in living cells. Because light production can be measured with high sensitivity and low background, luciferase-based assays enable the detection of very small changes in biological activity. This property led to the adoption of luciferase reporters in cell biology, pharmacology, toxicology, and environmental testing. Firefly luciferase, in particular, became a standard tool in laboratory research due to its dependence on ATP, which allows it to serve as an indirect indicator of cellular energy status.

Beyond basic research, luciferase has found applications in medical diagnostics and biotechnology. Luciferase-based assays are used to detect microbial contamination, measure cell viability, and quantify ATP in clinical and industrial samples. In vivo bioluminescence imaging, made possible by the expression of luciferase in living organisms, allows noninvasive monitoring of biological processes in animal models over time. This technique has become an important tool in cancer research, infectious disease studies, and drug development.

In summary, luciferase was discovered through the study of natural bioluminescence and established as a key enzyme responsible for light emission in diverse organisms. Its unique biochemical properties and the ability to generate measurable light under mild conditions have made luciferase an indispensable tool in modern science, with applications ranging from fundamental enzymology to advanced biomedical research.

References

de Wet JR, Wood KV, DeLuca M, Helinski DR, Subramani S (1987) Firefly luciferase gene: structure and expression in mammalian cells. Molecular and Cellular Biology 7 2 725–737 DOI: 10.1128/MCB.7.2.725-737.1987