Luciferase is a generic term for class of oxidative enzymes that produce bioluminescence , and is usually distinguished from a photoprotein. Name was first used by Raphael Dubois. Lucifer is a Latin word meaning Lightbearer. Luciferases are widely used in biotechnology, for bioluminescence imaging microscopy and as reporter genes Luciferases do not require an external light source, but do require addition of luciferin, the comsumable substrate.

1. Topic :
Luciferase
Submitted To,
Miss P.Mala
Associate Professor
Periyar Maniammai Institute of
Science and Technology
Submitted By,
Aleena Treesa Saji
123011356006
1st year
Biotechnology

2. Introduction
• Luciferase is a generic term for class of oxidative enzymes that
produce bioluminescence , and is usually distinguished from a
photoprotein.
• Name was first used by Raphael Dubois.
• Lucifer is a Latin word meaning Lightbearer.
• Luciferases are widely used in biotechnology, for bioluminescence
imaging microscopy and as reporter genes
• Luciferases do not require an external light source, but do require
addition of luciferin, the comsumable substrate.

3. Luciferase
• Luciferases are enzymes that produce light when they oxidize their substrate.
• The gene for the most common luciferase comes from the firefly, but luciferases
from other animals such as the sea pansy Renilla reniformis, the copepod Gaussia
princeps, and the ostracod Cypridina noctiluca are also used as reporters.
• When a luciferase is fused to a protein of interest, its expression can be measured
very accurately using a luminometer.
• The firefly luciferase reaction requires its substrate luciferin, plus adenosine
triphosphate (ATP), O2, and Mg2+.
• The Renilla and Gaussia luciferases use coelenterazine as their substrates;
Cypridina uses its own luciferin as a substrate.
• Luciferases are commonly used to report the expression level of proteins to
which they are fused.

5. Mechanism of reaction
• All luciferases are classified as oxidoreductases, meaning they
act on single donors with incorporation of molecular oxygen.
• Because luciferases are from many diverse protein
families that are unrelated, there is no unifying mechanism, as
any mechanism depends on the luciferase and luciferin
combination.
• However, all characterised luciferase-luciferin reactions to date
have been shown to require molecular oxygen at some stage.

7. Bacterial luciferase
• The reaction catalyzed by bacterial luciferase is also an oxidative
process:
• FMNH2 + O2 + RCHO → FMN + RCOOH + H2O + light
• In the reaction, molecular oxygen oxidizes flavin mononucleotide and
a long-chain aliphatic aldehyde to an aliphatic carboxylic acid.
• The reaction forms an excited hydroxyflavin intermediate, which is
dehydrated to the product FMN to emit blue-green light.
• Nearly all of the energy input into the reaction is transformed into
light.
• The reaction is 80% to 90% efficient. In comparison,
the incandescent light bulb only converts about 10% of
its energy into light and a 150 lumen per Watt (lm/W) LED converts
20% of input energy to visible light.

8. Biomedical application:
• Gene expression regulation
• Signal transduction
• Immunoassay
• Drug screening
• Molecular imaging based on cell detection
• Non-invasive in vivo imaging
• Protein protein interaction

9. MCQ - Question
• The name was first used by,
(a) Raphael Dubois (b)Raphael Davis (c)Raphael David (d)Davis
Raphael
• Meaning of Latin word lucifer,
(a)Fire bearer (b)Light bearer (c)Fruit fly (d)Dragon fly
• FMNH2 + O2 + RCHO →
(a) FMN + RCHO+ H2O + light (b) FMNH + RCOOH + H2O + light
(c)FMN + RCOOH + H2O + light (d) FMN + RCOOH + H2O
• Biomedical applications of luciferase
(a)Drug screening (b)Immunoassay (c)Both a and b (d)None of these
• All luciferase are classified as
(a)Oxidized (b)Reductase (c)Both a and b (d)Oxidoreductases

10. MCQ - Answer
• (a) Raphael Dubois
• (b) Light bearer
• (c) FMN + RCOOH + H2O + light
• (c)Both a and b
• (d) oxidoreductases

11. References
• Lee J (28 February 2014). "A History of Bioluminescence". photobiology.info.
Archived from the original on 29 March 2015.
• Williams TM, Burlein JE, Ogden S, Kricka LJ, Kant JA (Jan 1989). "Advantages of
firefly luciferase as a reporter gene: application to the interleukin-2 gene
promoter". Analytical Biochemistry.
• Nguyen VT, Morange M, Bensaude O (Jun 1988). "Firefly luciferase luminescence
assays using scintillation counters for quantitation in transfected mammalian
cells".
• Okamoto OK, Liu L, Robertson DL, Hastings JW (Dec 2001). "Members of a
dinoflagellate luciferase gene family differ in synonymous substitution
rates". Biochemistry
• "Introduction to Bioluminescence Assays". Promega Corporation. Archived from
the original on 2010-08-14.

12. THANK YOU