BIOLUMINESCENCE

Bioluminescence
What is bioluminescence

 Bio – living matter

lumin-to fill with light
escence-process that changes a state
biolminescence is the conversion of chemical energy
into light within a living organisms.

 Most marine bioluminescent organisms emit light in

the range of 440-480 nm ,which is conductive to
optical transmission through seawater.
Bioluminescent bacteria

Bacterial plate in the presence of light and absence of light.

Media

 Types of media:
 1.Luminescent agar:
 NaCl -10g
 Yeast Extract - 5g
 Peptone (Bacto-peptone) -10g
 Agar -15g
 Fill with H2O up till 1000 ml
 2. BOSS medium

 30 g NaCl
 1 g Glycerol
 10 g Peptone (Bacto-Peptone)
 3 g Beef Extract
 (to obtain solid medium you should add 15 g
of Bacto-Agar)
 Dilute the ingredients in water (pH should be
about 7,3)
 3. LM (Luminescence Medium)

 Yeast Extract 5 g
 Glycerol 3 ml
 CaCO3 (powdered limestone or chalk) 1 g
 Tryptone 5 g
 Sea water fill up till 1000 ml
What is Bioluminescence?

 The production of light by living organisms as a

result of a chemical reaction either within certain
cells or organs OR outside the cells in some form
of excretion.

 Produced by the compound luciferin and the

enzyme luciferase.
Who is lighting up in the ocean?
 Bacteria

 Dinoflagellates

 Jellyfish

 Squid & Octopus

 Annelids (worms)

 Krill & Shrimp

 Seastars, Brittlestars, Sea Cucumber

In the deep ocean, where sunlight is absent,
more than 90% of the animals are luminescent !!!

Krill
Ctenophore Deiopea
Comb Jelly
Annelid (segmented worm).
Lives 3000 m below surface.

Dinoflagellates
Small Squid

Enlargement of Photophores (light

emitting organs)
Vampire Squid
(Vampyroteuthis infernalis)
So….why do they do it?
Finding or attracting prey

 Fish such as the anglerfish use a light organ filled with bacteria that
dangles from their forehead.

 Prey are attracted to the light in the same way that a fisherman might
use a glowing lure for night fishing.

 Some fish use bioluminescence as a flashlight. They use light, produced

by symbiotic bacteria living in an organ below their eyes, to light up
potential prey.
Defence against predators
 Some squid and shrimp produce a luminescent glowing
cloud.
 When attacked by a predator, brittlestars sacrifice a part of
the body that continues to flash as the animal makes its
escape.
 Where the sunlight is dim, it can be used as camouflage.
 Many small plankton use flashes of light to escape from
predators.
 Communication / Mating
 Attract opposite sex.

 Bristle worms use bioluminescence during mating swarms, where the

males will attract females to them.

 Crustaceans send out coded messages when it is time to mate.

This octopod uses its bioluminescence as a private signal for mating.

Light is produced only at certain times, by a ring around the mouth of females.
Are bioluminescent animals
found only in the ocean?
 There are a few species on land.

 Fireflies during the summer in North America.

 Glow worms (actually fly larvae) in New Zealand.

 Railroad worms (actually beetle larvae) in South and Central

America.

 But, bioluminescence is not found in freshwater lakes

or rivers.
Chemistry of bioluminescent
bacteria
 Bioluminescence is light produced by a chemical
reaction within an organism.
Atleast 2 chemical reactions are required.

 Luciferin + oxygen-------> oxyluciferin +light

luciferase

1. The luciferase catalyzes the oxidation of

luciferin.
2. Resulting in light and an inactive oxyluciferin
 Firefly bioluminescence cycle

Luciferyyl adenylate

PPi Luciferase oxygen

ATP light

luciferin Oxyluciferin
Regenerating
reactions
• Bioluminescence is the emission of light from the
electron transport chain of certain living
organisms.
• Many marine microbes are capable of generating
bioluminescence.
• The enzyme luciferase picks up electrons from
flavoproteins in ETC and emits some of the
electronic energy as photons of light.
• The emission of light by luciferase enzyme
involves the presence of luciferin.
• The generation of light requires activation of
luciferin to form a luciferyl adenylate.
• The luciferyl adenylate is then acted upon by
molecular oxygen and luciferase to yield
oxyluciferin.
• This reaction which has intermediate steps is
accompanied by emmision of light.
• The color of the light flash differs with species
and strucure of the luciferase.
• Luciferin is then regenerated from oxyluciferin in
the subsequent series of reactions.
Benefits of Bioluminescence
Uses of bioluminescent

 1.Luciferin is used as a reporter gene and

inserted into a bacterial plasmid.

 2.Luciferin gene is added to cancer cells to

produce light.
 Genetics of biolumenescence
 Vibrio fischeri
 5 genes (lux CDABE) have been identified as active in
the emission of visible light.
 Lux R and lux L are involved in the regulating the
operon
The lux operon is a 9 kilobase fragment that controls
bioluminescence through the catalyzation of enzyme
luciferase
 Lux A and lux B code for the components of
luciferase.
 Lux C codes for the enzyme acyl –reductase.
i

Genetics of biolumenescence
 Lux D codes for enzyme acyl- transferase.
 Lux E makes the protein needed for the
enzume acyl protein synthetase.
Lux CDE codes for fatty acid reductase
complex.