Summary Sheet
Chemistry
Chemical Mechanism
Bioluminescence frequently gets confused with chemiluminescence, phosphorescence, and fluorescence
because they all have to do with chemicals producing light. However, bioluminescence is only related to
chemiluminescence, because it is a subset of this light producing method. Chemiluminescence is when
the energy that excites the particles in the substance to produce light originated from a chemical reaction,
and bioluminescence is chemiluminescence except the chemical reaction takes place inside an organism.
A minimum of two chemicals are required for this light reaction to take place, a light producing chemical
and a catalyst. The reaction can be triggered by neurological, mechanical, or chemical stimulus. The light
producing chemical is called a “luciferin,” and the catalyst is called a “luciferase” or “photoprotein.”
Luciferin would be the substrate in this light producing reaction, which is a substance that is affected by
an enzyme. To simplify the reaction, luciferin (obtained through diet or internal synthesis) is oxidized, the
oxidation driven by luciferase. Oxidized luciferin produces light, and “oxyluciferase,” which has no purpose
and is inactive. This reaction can be represented through this diagram:
Occasionally instead of producing light and oxyluciferase, luciferin, luciferase, and oxygen will bond to
create only one product, known as a “photoprotein.” This molecule will produce light when a specific ion,
commonly calcium, is combined with the system.
The words luciferin and luciferase have Latin roots, derived from the term lucifer, meaning “light bringer.”
Luciferin and luciferase are very broad terms, they are not the names of specific substances, but instead
are just a classification of different chemicals. Many substances can be luciferins and luciferases,
determined by the bioluminescent organism.
Luciferin and luciferase are proteins, meaning they are synthesized via the Central Dogma in organisms
through the formation of amino acids.
Physiology
Spectrum & Intensity
Most marine organisms give off blue light, emitting a wavelength that ranges from 440 nm to 479 nm.
Blue is the most common colour produced by bioluminescent organisms in marine environments because
blue light transmits the greatest through water, whereas all colours of light transmit well on land.
However, some organisms will create blue bioluminescent light, and then a green fluorescent protein
(GFP) will absorb those photons, and emit it closer to green light, around 505 nm.
Kinetics
Most bioluminescent creatures give light off in flashes, ranging from around a tenth of a second to ten
whole seconds. However, some can exude light in a continuous stream. Over a short period of time, many
dinoflagellates can react more than once when excitation occurs. In a majority of multicellular organism,
the neurological system directs bioluminescence.
Distribution
Bioluminescent creatures are located across the globe, however, there is a larger variety of creatures
located in the ocean (believed to be 80% of all luminous species). Dinoflagellates, which cause large
expanses of the ocean to glow, can be found in any ocean. However, they are more densely located
around areas that provide the most nutrients for their existence, such as bays and coral reefs.
DIVERSITY OF BIOLUMINESCENCE
Bioluminescent organisms span many kingdoms and phyla. There is no known pattern to the organisms’
distribution, they appear randomly across genera. Occasionally within a genus, some species are
bioluminescent and others are not, for which reasons can only be hypothesized. Bioluminescent species
are found in approximately 17 phyla and over 700 genera. Marine species include cephalopods, copepods,
amphipods, many fish, annelids, jellies, and many more. On land, there are insects such as fireflies, glowworms, and beetles, and well as different types of fungi. Bacteria that can bioluminesce occur on land and
in the water. Groups that lack bioluminescent members include: sponges, diatoms, mammals,
foraminiferans and heteropods. Bioluminescence occurs more frequently in species that live in deep
water, than those in benthic or shallow regions.
History
Ancient Era
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The first recorded instances of bioluminescent were made in 1500 to 1000 BCE.
They discussed the observation of firelfies and glowworms, however it is not known whether the
phenomenon was researched at all.
In the early CE, Pliny the Elder (23-79 CE), wrote the Naturalis Historia, which contained
comprehensive information about numerous bioluminescent organisms, including, glowworms,
firelfies, and luminous mollusk.
The mollusk was named Pholas dactylus, named by the Romans, to whom it was a delicacy.
Middle Ages to the Sixteenth Century
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The church controlled the acquirement of knowledge, and bioluminescence was not allowed to
be studied because it was believed to be magic
Seventeenth Century
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The seventeenth century was dominated by trying to find and recreate the Philosopher’s stone,
which is said to turn materials into gold and provide immortality.
In the late seventeenth century however, Robert Boyle (well known for establishing Boyle’s law)
conducted experiments on the effects of oxygen in living and nonliving substances. He explained
that oxygen is required for respiration in living animals, as well as for bioluminescence (Roda,
2010).
Eighteenth Century
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The scientist Spallanzani from Naples, Italy, carried out experiements on bioluminescent medusae
in 1797. These experiments proved that oxygen, water, and a specific photogenic substance are
required for bioluminescence, as well as showing that the production of light is not necessary to
the organisms survival (Roda, 2010).
Many scientists throughout the eighteenth century deduced mechanisms for light production in
organisms, such as De Mairan stating that the bioluminescence in jellyfish could be caused by the
movement of sulfur particles, and Benjamin Franklin suggested that bioluminescence could be
caused by electricity (Roda, 2010).
Twentieth Century
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The twentieth century yielded massive advancements in the study of bioluminescence, due to the
development of new scientific instruments, such as photomultipliers, imaging tubes, and lasers.
The chemical mechanisms and the numerous types of luciferins were identified, as well as the
structure of many different luciferase enzymes were discovered.
J. Woodland Hastings research on biolumminescence also led to the discovery of green
fluorescent protein in the later 20th century.
Evolutionary Advantages
Startle
There are more known cases of organisms using bioluminescence for defense than offense. When a
predator is within range of a bioluminescent organism, the organism can emit a flash of light. This scares
the predator and can cause them to delay their attack, possibly giving the prey enough time to escape.
Misdirection
When bioluminescent particles are secreted, the cloud emitted can screen the prey from the predators.
This allows for the prey to escape, and the predator will not be able to see it to follow.
Sacrificial Tag
Another defense mechanism is also the use of a sacrificial tag. A sacrificial tag is when an organism will
detach a luminescent part of its body, attracting the predator to that detached member, and the predator
will often eat the tag instead of the prey. The tissues can emit light for long periods of time after it has
been detached, even when it has been consumed by a predator. This will cause transparent predators to
emit light, which can draw unwanted attention.
Counterillumination
Counterillumination is a type of camouflage most frequently displayed in crustaceans, cephalopods, and
fish. It uses light emitting organs on the bottom of the organism to get rid of the shadow that the organism
would cast.
Burglar Alarm
The burglar alarm method works similar to the sacrificial tag, the organism that is being preyed upon will
emit light either internally or externally, which will draw attention to the predator. This can attract
secondary predators to the situation.
Aposematism
In terrestrial situations, it is known that bright colours on an organism demonstrates that it is not safe to
eat. This is true for terrestrial bioluminescent organisms as well. However, it is not known whether marine
predators recognize aposematism.
Offense: Prey Attraction
Bioluminescence can be used in the same way in which a moth is attracted to a flame. Organisms,
frequently fish, will use their glowing abilities to attract prey, and then they will capture the organism
when it comes within a suitable distance.
The emission of intense light can also be used to stun prey. Many organisms will produce light while
attacking their prey, and others have forward pointing lights. These forward pointing lights may also be
used for lighting up prey, or making them emit fluorescence.
Reproduction
Many organisms use bioluminescence to communicate among their species, or to attract mates. Patterns
in light emissions can also allow for organisms to identify each other (Haddock, 2010).
Bioluminescent mushrooms use their light to attract insects to land on them, which helps to spread their
spores.
Metabolic By-product?
Bioluminescence may have many functions, but it is likely that in bacteria, light emission was just a
metabolic by-product. Oxidation reactions would take place in the cells of the organism, and luciferins
may have been present because they are an antioxidant. Therefore, the presence of the molecules that
would eventually create bioluminescence were beneficial before their evolutionary defense and offense
functions became beneficial.
Applications
BIOASSAYS
A bioassay is an experiment that is done to measure concentration, potency, or toxicity of a substance
using living things (Cornell University, 2009). Biological activity can be inhibited by many manufactured
substances, such as anesthetics, narcotics, factory poisons, insecticides, pesticides, and medical
substances, to which these substances bacterial bioluminescence has an extraordinary sensitivity.
Therefore, bioluminescent cells can be used as bioassays.
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Single Luciferase Reporter Assay
Dual Non-Secreted Luciferase Reporter Assay
Dual Secreted Luciferase Reporter Assay
Multicolour Luciferase Reporter Assay
Calcium Ion and ATP Monitoring
Calcium and ATP levels can be monitored inside a cell using a dual-colour system, through a red luciferase
from a beetle that is ATP sensitive, as well as a calcium sensitive aequorin.
Bioluminescence Imaging
The simplest and cheapest procedure for molecular imagining uses bioluminescent or fluorescent
reporters. Images of single molecules or even entire organisms can be sampled using a fluorescent dye
molecule or a fluorescent protein that has been genetically encoded. This has led to a massive
advancement of cell biology. However, fluorescent reporters require an external source of illumination to
produce light, and therefore this technique cannot be used for long-term or quantitative imaging. The
fluorescent reporter can be bleached by the external lamination, or it can cause damage to the cells and
light sensitive tissues. Bioluminescence imaging can be used instead because it does not require external
light sources.
Glowing Plant
Glowing Plant is a company that wants to create glowing plants that can be substituted for table or desk
lamps, which require no energy input.
How They Make the Glowing Plant:
The plant is created using a design, print, and transform cycle; mechanisms of synthetic biology. Using
Biocad software on a computer, they create DNA sequences that contain information for production of
bioluminescence, and the DNA is then laser printed. The DNA is then inserted into the DNA of the target
plant using a Gene Gun, fabricating a plant that can generate its own light.
Dino Pet
The Dino Pet is a decoration that can be purchased online. It is a clear dinosaur shaped aquarium that can
is filled with live dinoflagellates.