the topic describes about the luminescence, its presence in different microbes, the luminescent beaches in world and useful aspects of this phenomenon

1. Bacterial
Bioluminescence
Presented by
SARA ISHAQ
IQRA MALIK
IRUM TEHSIN

2. Bioluminescence
• Phenomenon of production and emission of
light by an organism through chemical
reaction.
• Chemical energy Light energy
• Bioluminescent bacteria
Are mostly----Marine
Few are --------terrestrial

3. Aquatic Bioluminescence
• 80-85%of oceanic world is bioluminescent.
• Jelly fish, coral
• Din flagellates

4. Terrestrial bioluminescence
• Fungi , Algae
• Mushrooms
• Firefly
• Glow Worms
• Earthworms

5. List of Bioluminescent
Organisms
• Terrestrial animal:
 Certain anthropoid
 Fire flies
 Click beetle
 Glow worms
• Marine animals:
 Anglerfish
 Flashlight fish
 Black dragon fish
 Sparkling enope
squid

6. • Fungi:
 Armillaria calvescens
 Armillaria gallica
• Bacteria:
 Photobacterium
 phosphoreum
 Photorhabdus
luminescens
• Protists:
 Pyrodinium bahamense
 Lingulodinium
polyedrum

7. Family Genus Species
Enterobacteriaceae Photorhabdus
Photorhabdus asymbiotica
Photorhabdus luminescens
Photorhabdus temperata
Shewanellaceae Shewanella
Shewanella woodyi
Shewanella hanedai
Vibrionaceae
Aliivibrio
Aliivibrio fischeri
Aliivibrio logei
Aliivibrio salmonicida
Aliivibrio sifiae
Aliivibrio "thorii"
Aliivibrio wodanis
Photobacterium Photobacterium aquimaris

8. Lights on

9. Lights off

10. At very first Bioluminenscence were recognized in 180 marine
species known as “cold light”
In 15th century commented these luminance in bacteria as
“Burning sea”
In 16th century bioluminescence found in small literature commonly
called “unaffected fire”
in 17th century observe that air is required for luminance not
oxygen. but in reality oxygen is required.
Later in 19th century extracted the two key points which causes the
bioluminescence reaction are” lucifirine” and “luciferase”.
HISTORY

11. Genetic diversity
• All bioluminescent bacteria share a common
gene sequence;the lux operon characterized
by the luxCDABE gene organization.
• Based on similarities in gene content and
organization lux operon can be organized into
four distinct types.
3:vibrio/
candiddtus
1:Aliivibrio/shewa
nella
2:Photobacterium
4:Photodesmus

12. Commonly occurring types
1: Photobacterium and
their species are;
• Photobacterium
aquimaris
• Photobacterium
mandapamensis
• Photobacterium
phosphoreum
2: Vibrionaceae(vibrio)
• Aliivibriofischeri
• Aliivibriologei
• Aliivibrio sifiae
• Aliivibrio thorii
• Aliivibrio wodains

13. Shapes……
Photobacterium
species
vbrionaceaei

14. Beaches containing bioluminescence
• There are five bioluminescent bays in the
world.
1: Luminous lagoon in Jamaica
2:Halong in vietnam
3:Puerto Rico’s Laguna Grande
4:Laparguera
5:Mosquito Bay
Mosquito bay is currently the brightest.

15. Luminois lagoon
Halagoon
3:Puerto Rico’s
Laguna Grande
5:Mosquito Bay

17. Symbiotic bioluminescence bacteria
1:to obtain food
• Most species of luminescent bacteria are
capable of living free in association with host
organisms to obtain food and protection.
Pinecone fish utilize luminous
bacteria, colonizeed in the ventral
cavity,illuminate surrounding.

18. 2:Attract the prey
• In symbiosis bacteria nourished with available
food for growth.at same time host utilizes the
adopted illumination to attract prey.
Deep sea Angler fish carries
the luminance bacteria.

19. 3:Mutualistic association
• In this association both host and luminated
bacteria are benefited.
• Luminous bacteria in symbiosis on a pair of light
organs in the mantle body of the squid.
utilize the luminated function causes to frighten
the nearby predators.

20. Why does it occur !!
Invitation to a
meal
Puzzling predatorsClever disguise
Mating games Mating gamesDefense

21. Biochemistry of the Bacterial
Bioluminescence Reaction-
Bacterial Luciferase and the
Light Color

22. Bacterial luciferase
 The enzyme that catalyzes light emission
 Heterodimer
 Composed of two different polypeptides,
designated alpha and beta and encoded by
the luxA and luxB genes, respectively.
 The active site is located
within the subunit.

23.  In the absence of the beta subunit, the alpha
subunit alone functions inefficiently with a
poor light yield.
 However, the catalytic machinery involved in
continuous light production in luminous
bacteria includes not only bacterial luciferase,
but also the enzymes that supply and
regenerate the substrates of bacterial
luciferase.

24. Lux Genes
The DNA sequences coding
the proteins in the
luminescent system are
termed the lux genes.

25. Substrates of Bacterial
Luciferase
1. Reduced flavin mononucleotide (FMNH2)
2. Molecular oxygen
3. Long chain fatty aldehyde

26. Biochemical Mechanism
 The reaction of bacterial luminescence is catalyzed by
luciferase.
 The reaction leads to
1. the oxidation of FMNH2 to FMN
2. the oxidation of the aldehydes to organic (fatty) acids
3. . A quantum of light is an additional product of this
reaction

27. The net chemical equation of the
bacterial luciferase catalyzed reaction.
RCHO + FMNH2 + O2 → RCOOH + FMN+H2O + hν

28. The excess energy, which is liberated from the
oxidation of FMNH2 and aldehyde concomitant
with the reduction of molecular oxygen, is
released as blue/green light emission (MAX ~
490 nm).

29. Different Luciferase Emission
Colors.
 The characteristic color indicates the energy
level of the photon that was produced when
the excited electron on the flavin
chromophore returns to the ground state.

30. 1. Flavin analogs
 Flavin analogs with substituted atoms in the
chromophores moiety resulted in different
luciferase emission colors.

31. 2.Point mutations
Point mutations at the flavin chromophore's
binding site distorts the color emission spectrum
of bacterial bioluminescence, indicating that the
distinctive emission color depends not only on
the chromophore that emits the photon, but also
the electronic nature of the chromophore-binding
microenvironment in luciferase.

32. 3. Fluorescent proteins
some luminous bacteria carry fluorescent
proteins to modulate the emission color,
distinguishing themselves from other strains.

33. How Does Bioluminescence
Work?
• Bioluminescence is a product of chemical reaction in
an organism.
• It involves a class of chemical called luciferins (light
bringers).
• The luciferins oxidizes in the presence of a catalytic
enzyme(luciferase) to create light and an inactive
compound(oxyluciferins).

34. HOW DOES IT WORK?
• In bioluminescence, a
luciferin produce lights & a
luciferase the light producing
chemical reaction to take
place
• In this reaction luciferin act
as a catalyst.
• Luciferase allows oxygen to
combine with luciferin
• The reaction produces
photons of light
• And oxidized luciferin
becomes inactive oxyluciferin

35. What is the Difference Between Bio fluorescence
and Bioluminescence?
Bioluminescence
 Bioluminescence is a
chemical process in
which an enzyme breaks
a substrate down and
one of the products of
this reaction is light.
 The most popular usage
of luciferase (an enzyme
that causes
bioluminescence in
fireflies and sea pansies)
is to test that activity of
gene regulatory
elements
Biofluroscence
 Bio fluorescence is a
physical process by
which light excites
electrons in the
fluorophor to a higher
energy state, and when
that electron falls back
down to its ground state
it emits a photon.
 The likelihood of
measuring auto
fluorescence or
excitation photons is
extremely low

36. How Can We Make Use of Bioluminescent
Chemical for Our Own Benefit?

37. Bioluminescence Modern Day
Application Biology and medicine:
1. Luciferase systems are widely used in
genetic engineering as reporter genes.
2. Bioluminescent activate destruction is an
experimental cancer treatment
3. Vibrio bacteria symbiosis with marine
invertebrates such as the Hawaiian bobtail
squid are key experimental models for
bioluminescence.
4. Its used for bio monitoring.

38. In Environment:
1. Detection of drugs in surface water and
waste water samples preliminary testing of
toxicity.
2. Assessment of heavy metal by bacterial
bioluminescence in waste water.
3. Dinoflagellates bioluminescence for
environment risk detection.
4. Detection of specific pollutants in
environment.

39. In Industrial field:
Structures of photophores, the light
producing organs in bioluminescent
organisms, are being investigated by
industrial designers.

40. Others field:
1. Engineered bioluminescence could
perhaps one day be used to reduce the
need for street lighting.
2. It also used in energy consumption.

41. Fluorescence microscopy
•Fluorescence microscopy of tissues, cells or
subcellular structures is accomplished by
labeling an antibody with a fluorophor and
allowing the antibody to find its target antigen
within the sample.
• Labeling multiple antibodies with different
fluorophores allows visualization of multiple
targets within a single image.

42. Automated sequencing of DNA
•Automated sequencing of DNA by the chain
termination method; each of four different chain
terminating bases has its own specific
fluorescent tag.
•As the labeled DNA molecules are separated,
the fluorescent label is excited by a UV source,
and the identity of the base terminating the
molecule is identified by the wavelength of the
emitted light.

43. Bioluminescence imaging (BLI) is a technology
developed over the past decade that allows for the
noninvasive study of ongoing biological processes
in small laboratory animals.
Common applications of BLI include in vivo studies
of infection (with bioluminescent pathogens),
cancer progression (using a bioluminescent cancer
cell line), and reconstitution kinetics (using
bioluminescent stem cells).
BLI

44. BRET: Bioluminescence Resonance
Energy Transfer
• BRET is a proximity-
based assay where the
energy generated by the
catalytic degradation of
coelenterazine by the
enzyme Renilla
luciferase (Rluc)
(energy donor) is
transferred to a green
fluorescent protein
(GFP) acting as the
energy acceptor.
• The GFP then emits
light at its specific
emission wavelength.

45. BRET

46. BRET: Bioluminescence Resonance
Energy Transfer
• BRET can be used to observe protein-protein
interaction in living mammalian cells.
• It is based on the non-radioactive transfer of
energy between a luminescent donor Rluc
and a fluorescent acceptor (Green
Fluorescent Protein or GFP).
• May be used in the future to identify new
protein complexes in human.

47. Monitoring of ubiquitination in living
cells by BRET
• Ubiquitination is descried as the rapid
process of post-translational modification
present in many aspects of biology involving
a covalent attachment of ubiquitin to
proteins.

48. Lux operon
• After the discovery of the lux operon,
the use of bioluminescent bacteria as
a laboratory tool is claimed to have
revolutionized the area of
environmental microbiology.

49. Biosensors
• The applications of bioluminescent bacteria
include biosensors for detection of
contaminants, measurement of pollutant
toxicity and monitoring of genetically
engineered bacteria released into the
environment.
• Biosensors, created by placing a lux gene
construct under the control of an inducible
promoter, can be used to determine the
concentration of specific pollutants.

50. • Biosensors are also able to distinguish
between pollutants that are bioavailable and
those that are inert and unavailable.
• For example, Pseudomonas fluorescens has
been genetically engineered to be capable of
degrading salicylate and naphthalene, and is
used as a biosensor to assess the
bioavailability of salicylate and naphthalene

51. Gene expression
• Bioluminescence can be used to study
prokaryotic gene expression inside
living cells
• It allows the observation of biological
processes in real time, as they happen.
• This technique can be used as a
noninvasive way to study protein
trafficking, protein function, genetic
regulatory or image bacteria, tumors
and genes over a long period time.

52. GENETIC ENGINEERNING
Genetically engineered Angelfish (Pterophyllum Scalare) glow
in a tank under a black light while being displayed at the 2010
Taiwan International Aqua Expo in Taipei October 29, 2010.

53. Applications of bioluminescent imaging.
(a) assessing the levels of trans-gene expression,
(b) the location and extent of bacterial infection,
(c) the efficiency of gene transfer and expression,
(d) the trafficking patterns of lymphocytes.

54. Bioluminescence in gene
expression
• Luciferases are
light-generating
enzymes that can
be found in
bacteria, marine
crustaceans, fish
and insects.
• Luciferases are
nontoxic and can be
injected to become
gene expression
markers.

55. Gene Expression in Yeast Cells
• Cells fused to GFP,
making the component
protein of microtubules
(alpha-tubulin)
• The alpha-tubulin:GFP
fusion can be observed
by exciting the GFP,
causing a green light
emission.
• It can be used to study
gene coding mutations
such as kinases.

56. Bioluminescence in Food
Industry
• Methods of use
ATP
bioluminescence
• Bacterial
bioluminescence

57. ATP Bioluminescence
• Measures the amount of ATP that is
converted to photons of light by living
cells.
• The amount of light emitted to
proportional to the number of bacteria
in a food sample
• It can be used to measure the number
of lactic acid bacteria in a
contaminated food sample.
• Firefly luciferase is used to detect the
presence of ATP

58. Luciferin and the reaction
with ATP

59. The visible glow of pathogen
helps provide instant counting
results.
PATHOGEN

60. Detection of Bacteria

61. Military
Biodegradable landing zone markers
Bioluminescent "friend vs. foe"
identification markers and security
systems

62. Large Scale Production Setup
Production
center for
Luciferase
Enzyme
Research
Development
Commercialization
Can be located near areas like
Food contamination test
centers .
Requirement for
Bioluminescent based lighting
Can extract the enzyme in
large scale.
Production of synthetic
Luciferin
Can be used to make
Bioluminescent based lighting.
•Can employ “Made to order”
lights / artistic pets / wall
design / Bill boards and
Biosensors

63. Artistic Inspirations
• Poetry
• Photography
• Painting

64. Still Photography
• Artistic recreation of the
Bioluminescent Bay in Vieques

65. Sculpture
• Montana State University-Bozeman
Bioglyphs project
• Collaboration between art and science
during 2002 by members of the center
for Biofilm Engineering and the MSU
School of art

66. Bioglyphs
• Involves the practice of "painting" on
prepared Petri dishes with a sort of "invisible
ink" composed of liquid medium inoculated
with the bacteria.
• The microorganisms themselves went to
work, multiplying on the plates and beginning
to produce light within 24 hours.
• The only light available to view the art was
that produced by the bacteria themselves.
• Over the five-day period, the light intensity of
the paintings changed as the bacteria
multiplied and then gradually consumed the
nutrient available.

67. • Extensive research is going on
whether the plants are able to show
bioluminescence.
• Daan roosegaard’s team has recently
declared by merging their light
producing compound with plants, the
team envisions illuminating city
streets with trees that glow at night.

68. A university of Cambridge team modified genetic
material from Fireflies and the luminescent
bacterium vibrio fischeri to boost the production of
light-yielding enzymes that can ultimately be
inserted into genomes – they called it biobricks.
The team generated two lines of common
nicotiana tabacum houseplants that carried the
bacterial lux operon from photobacteriu leiognathi.

69. As a result, the plants can produce luciferase
and their substrates,
Luciferinsengineered bioluminescence could
perhaps one day be used to reduce the need
for street lighting or decorative purposes

70. Scientists are researching the use of
genetically engineered bioluminescent.
E. Coli for use in a bio bulb the gene that
makes firefly’s tails glow has been added
to mustard plants.
The plants glow faintly for an hour when
Touched, but a sensitive camera is
needed to see the glow.

71. • In Vivo Imaging
• Detection of key Diseases
• Oncology/Cancer
• Inflammatory Diseases
• Neurology
• Cardiovascular
• Drug Metabolism Studies
Other Application
• Monitoring Treatment
Response
• Biodistribution
• Cancer cell detection
• Biomarkets
• Structural Imaging

72.  Nanotechnology
System on Chip
Nano particles
Nano polymers
Minimum flashing
Maximum
Luminosity
 MEMS
Computational DNA‟s
 Genetics
DNA improvements
Technology merge
BIOLUMINESCENT
SYSTEM ON CHIP
Technology Convergence

73. THIS IS WHAT OUR
STREETS WILL LOOK LIKE
IN THE FUTURE….

77. Lights on

78. Lights off

79. Gallery, with lights on

80. Gallery, with lights off

83. •DO u have any Questions
please????

84. Sources
• Stephane Angers, Ali Salahpour, Eric Joly, Sandrine
Hilairet, Dan Chelsky, Michael Dennis, and Michel
Bouvier. “Detection of β 2- adrenergic receptor
dimerization in living cells using bioluminescence
resonance energy transfer (BRET).” Proceedings of the
National Academy of Sciences 97(2000): 3684-3689.
http://www.pnas.org/cgi/content/full/97/7/3684 (5
October 2007)
• Claire Normand1, Stéphane Parent1, Benoit Houle, Anne
Labonté, Lucie Bertrand, Mireille Caron, Mireille Legault,
Stéphane Angers, Michel Bouvier, Erik C. Joly and Luc
Ménard. “BRET2™: Bioluminescence Resonance
Energy Transfer, a Novel Assay Technology to Examine
GProtein Coupled Receptor Activation in Intact Cells.”
2002
http://las.perkinelmer.com/Content/RelatedMaterials/Po
sters/PSH_BRET2NovelAssayTechnology.pdf (9 October
2007).