Bioluminescence and its Applications and Economic Feasibility

These slides use concepts from my (Jeff Funk) course entitled analyzing hi-tech opportunities to analyze how the economic feasibility of bioluminescence is becoming better through finding better sources of bioluminescence and reducing the cost of Luciferin & luciferase. Organisms displaying bioluminescence include fungi, algae, mushrooms, fireflies, glow worms, earth worms, and jelly fish, coral, to name a few. An astonishing 80-85% of the deep oceanic world is bioluminescent and some of this can even be seen from outer space! Bioluminescence primarily occurs when chemical Luciferin reacts with oxygen in the presence of catalyst luciferase.
There are some exciting things about bioluminescence. First, it is possible to achieve it even with a coating as thin as 1mm as it works at microbial level. This would further reduce the size of lights far beyond what is possible with LEDs and OLEDs. Second, it is theoretically possible to grow trees and plants that are bioluminescent and thus use them for lighting streets and other outdoor areas. Clearly this would be tremendous success and have a large impact on the world’s energy needs in addition to possibly improving the aesthetics of cities.
More immediate applications can be found in disease detection where Bioluminescence is already being used. Specific cells, viral agents, or genes can be bioluminescently labeled. After injecting them into an organism, cameras and spectral analysis can be used to detect their movement and multiplication. This can be potentially much cheaper than MRI, computer tomography and other approaches; the challenge is to create the different bio-luminescent materials(synthetic Luciferin), their spectral signatures, and methods of injection.
A final challenge that is common to all bio-luminescent applications is the cost of the biological material, which is usually Luciferin. Extracting it from fireflies can cost tens of thousands of dollars for few milliliter, clearly a very high cost. However, synthetic methods of production have been devised and scaling up these production plants will likely lead to much lower costs. Given the experience in the chemical industry it is likely that in future with advances in technology the costs may fall more than a thousand times as the production processes are scaled up to the levels found in high-volume chemicals.

1. Bioluminescence
MT5009 – Analyzing Hi-Technology Opportunities

2. 2

3. Bioluminescence
3

5. 5
Bioluminescence Why does Bioluminescence occur?
How it Happens ?
Bioluminescence Vs. Fluorescence Vs.
Incandescence
Opportunities Lighting
Biomedical Imaging
Food Industry
Inferences and
Conclusion
Commercialization
Entrepreneurial Opportunities
Technology Convergence
Conclusion
Outline

6. 6
Terrestrial Bioluminescence
Firefly
Glow Worms
Earthworms
Fungi, Algae
Mushrooms

7. •80- 85% of oceanic world is
bioluminescent
•Jellyfish, coral.
•Dinoflagellates
7
Aquatic Bioluminescence

8. 8
Bioluminescence Near Cape Horn

9. •Chart with sea depth and light penetration
Major reason why most bioluminescent organisms emit blue or green color
9
Bioluminescence.. In Sea

10. Invitation to a meal
Mating games
Puzzling predators
Burglar Alarms
Clever disguise
10
Why does it occur !!

11. How it Happens?
11

12. Bioluminescence Fluorescence Incandescence
Lowest efficiency
12
Bioluminescence Vs. Fluorescence and
Incandescence
Light Light Heat Light Chemical Light
Moderate Efficiency Highest Efficiency

13. Bioluminescence
Bioluminescence Vs. Fluorescence and
Incandescence
13
Source: STRUCTURE and SPECTRA in BIOLUMINESCENCE John Lee
1
and Eugene S. Vysotski
1,2

1
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602
Bioluminescence has a spectrum from 430 nm to 600 nm

14. Parallel Streams of Research
14
CAN be Engineered
CANNOT be Engineered
Focus on optimizing growth conditions

Study on lighting patterns to improve collective flashing

Bioluminescent Dinoflagellates
Synthetic Biology

Bioluminescent bacteria

Development of synthetic Luciferin

Vibrio Fischeri animation

15. Applications Lighting
hito ni akali wo nlisuru hotaru kana
The firefly Gives light to its pursuer…
15

16. Lighting Industry
Source: Lighting the way: perspectives on global lighting market 2012, Mckinsey study
16
Lighting consumes a substantial amount of electricity

17. 17
Lighting consumes most Electricity
According to EIA, in commercial buildings Lighting fixtures
consume 3 times the energy consumption of air conditioning

18. Incandescence is “Hot light” or black body radiation

Creates light by heating of filament

Highly inefficient! 90% of energy is wasted to heat &
UV

Adverse impact on environment- 69% of the energy
used in coal, natural gas and nuclear power generation
never does useful work as electricity

Short lifespan- 1000 to 1500 hours

Susceptibility to shocks and vibrations
Problems with Incandescence
18

19. Cold light

Most efficient of all the sources of lights. ~98% efficiency

As cultured at micro-organic level, immune to external impacts

No environmental impact

Once genetically re-engineered, unlimited source

“Grow your own light“
Bioluminescence as solution..
19
Why is Bioluminescence special ?

20. Uses bioluminescent bacteria, fed on methane and composted material

Provides soft mood lighting

Produces low intensity lighting, suitable for tracing, warning, indication

In present form not suitable for functional illumination

Luminescence by using energy stored in waste streams

POC of a self sustainable microbial lighting system
Current Research: Philips Concept
20
Cultured Bacteria having
conducive environment
Source: http://www.design.philips.com/philips/sites/philipsdesign/about/design/designportfolio/design_futures/bio_light.page

21. Team of researchers from Cambridge placed genes from fireflies and
bioluminescent bacteria into E.coli

Codon optimization and single amino acid mutagenesis allowed us to
generate bright light output in a range of different colors

Proposed a genetically modified tree which can be source of light in the
dark

Biological alternative to conventional lighting

Potential could replace street lamps
Current Research: Cambridge Concept
21
Source: http://www.holcimfoundation.org/T1560/Bioluminescent_lighting.htm

22. •Thinkers at IDEO are working with scientists from Lim Lab at the University of
California, San Francisco to find a way to have E. coli bacteria form objects.

•Like 3D printing, idea is to literally “grow” the product we use everyday. Even
lights !!!

•Idea is to put genes into an organism to make them self-aggregate into a solid
material with the desired properties.

Current Research: IDEO Concept
22
Source: http://www.fastcompany.com/biomimicry/training-bacteria-to-grow-consumer-goods

23. •Luminosity comparison
–Spectrum of bioluminescence almost similar to fluorescence though its
more spread out.







•Scalability of the light source- multiplication of bacteria.
–Today scientists are able to synthetically culture genetically modified
bacteria.
–Research in substrate for repetitive culture.

Dimensions of Performance
23

24. •Synthetic Luciferin Vs. Natural Luciferin gene
–Scientists have been able to invent synthetic chemicals which have
almost similar relative light Unit output to firefly gene.









Dimensions of Performance
24

25. •Luminosity of Bioluminescence
Limitations of Bioluminescence based
Lighting
Light source Output (Lumens)
Firefly 0.04
Incandescent 200-800
Fluorescent 1000-7500
Metal Halide 1900-30000
High-Pressure
Sodium
3600-46000
Low-Pressure
Sodium
1800-33000
25
~25,000 fireflies to flash
simultaneously to match output of
60W bulb !!!
Flashing









Better understanding of the chemistry of
bioluminescence

Better control on flashing and are able to minimize
it by studying fireflies.

26. Comparisons

27. 28
Technology Adoption
High
Medium
Low
Technology Comparison
Drivers of Adoption

28. 29
Bioluminescence Why does Bioluminescence occur?
How it Happens ?
Bioluminescence Vs. Fluorescence Vs.
Incandescence
Opportunities Lighting
Biomedical Imaging
Food Industry
Inferences and
Conclusion
Commercialization
Entrepreneurial Opportunities
Technology Convergence
Conclusion
Outline

29. Radiation and Scans

30. In Vivo Imaging
A noninvasive insight into living organisms

Understand disease related changes in the body
What is In Vivo Imaging ?
70%
36% 32% 25% 18%
Oncology/Cancer Inflammatory
Diseases
Neurology Cardiovascular Drug Metabolism
Studies
Detection of key Diseases
Other Application
68%
59% 49% 41%
24%
Monitoring
Treatment Response
Biodistribution Cancer cell
detection
Biomarkets Structural Imaging
Source : Drug Discovery World Summer 2011, In vivo preclinical imaging Layout 1 23/06/2011
14:05 Page 59

31. In Vivo Imaging
Conventional Methods
Magnetic Resonance Imaging (MRI)
Single photon emission computed
tomography (SPECT)
Positron emission tomography (PET)
MRI
SPECT
PET
Optical Imaging – Bio Luminescence Imaging
Optical,
28%
MRI, 23%
PET, 20%
SPECT, 3%
Others,
26%
Optical MRI PET SPECT Others
Source : Drug Discovery World Summer 2011, In vivo preclinical imaging Layout 1 23/06/2011
14:05 Page 59

32. Bacteria Cell
Virus Cell
Transgene
Bioluminescent Cell /
Receptor Gene
BLI - Concept
1 2 3 4
Inject, Infect, Implant Image Acquisition Data Analytics
1
2
3
4
Bacterial Cells, Viral agents or genes can be bioluminescent labeled
Labeled cell or gene is implanted into a mouse, Cells are allowed to
multiply
Mouse is placed highly sensitive CCD camera enclosure to obtain a 3D image
Spectral analysis is used to see the progress of the malignant cells
Source : A New Diagnostic System in Cancer Research: Bioluminescent Imaging (BLI)*, Z., Ralph MASON, Peter ANTICH,
Edmond RICHER, Woodring E. WRIGHT

33. BLI - Concept
Growth of cancer cell over 4 weeks can be observed
Source : A New Diagnostic System in Cancer Research: Bioluminescent Imaging (BLI)*, Z., Ralph MASON, Peter ANTICH,
Edmond RICHER, Woodring E. WRIGHT

34. PET vs. MRI vs. BLI
Equipment Costs
Mouse Preparation
Time
Scanning Time
Total Time for
1 animal
Data Analysis
Total Time for
10 animal
>600 K USD 1-2 million USD <500 K USD
1hr 30 min 30 min 20 min
15min / 3D Scan 30 min / 2D Scan 1s – 2min
Experts Required Experts Required Straight Forward
1 hr 1hr 30min 20min
>600 K USD 1-2 million USD <500 K USD
Data Analysis Time
>600 K USD 1-2 million USD <500 K USD
PET MRI BLI
Source : A Comparison of Imaging Techniques to Monitor Tumor Growth and Cancer Progression in Living Animals, Anne-
Laure Puaux,Lai Chun Ong,Yi Jin,Irvin Teh,Michelle Hong,Pierce K. H. Chow, Xavier Golay

35. Market Potential – In Vivo Imaging
790
900
1050
1200
1350
1550
0
200
400
600
800
1000
1200
1400
1600
1800
2012 2013 2014 2015 2016 2017
$ Million
Source : WWW.Marketresearch.com
CAGR
14.5%

36. Cost Radiation
Level
Scanning
Time
Imaging
Quality
Preparation
Time
Bioluminescence MRI PET
Low
Medium
High
Technology Comparison

37. Main Drivers for Technology Adoption
40%
40%
20%
Efficacy Health Safety Cost

38. 39
Bioluminescence Why does Bioluminescence occur?
How it Happens ?
Bioluminescence Vs. Fluorescence Vs.
Incandescence
Opportunities Lighting
Biomedical Imaging
Food Industry
Inferences and
Conclusion
Commercialization
Entrepreneurial Opportunities
Technology Convergence
Conclusion
Outline

39. Food Industry
•Current food regulations
Food development and Authority
HACCP (Hazard Analysis and Critical Control
Points)
Principle 4 – Establish monitoring procedures
When and which food should undergo
microbiological test
A study by the Leopold Center for Sustainable
Agriculture in The USA showed a 22% increase in the
average distance travelled by food products (arriving
in Chicago) by truck in the past 2 decades.
Food travels longer distances today
Source:1) http://www.fda.gov/Food/GuidanceRegulation/HACCP/HACCPPrinciplesApplicationGuidelines/default.htm
2) http://www.landcareresearch.co.nz/__data/assets/pdf_file/0003/39927/food_miles.pdf

40. According to The US National Library of Medicine, in the United
States alone about 48 million people get sick from consuming
contaminated food each year.

41. Existing method for detection of
food contamination
Sample from
food item
Dilution planted in
agar based media
Incubation period Counting bacteria
Total Viable Count (Standard Plate Count/Aerobic Plate Count)
Source : Essential Microbiology for Pharmacy and Pharmaceutical Science, By Geoff Hanlon & Norman Hodges

42. Existing methods for detection of
food contamination - Limitations
Expensive laboratory equipment.
Requirement of specialized transportation.
Long wait involved, anywhere between 24-48 hours. In cases of fermented foods
(such as soy sauce) this period could extend up to 7 days. This increases the storage
time before fresh food can reach the market, and may actually decrease the quality
of the food during the waiting period.
Tedious and labor intensive. The enumeration of colonies is performed using an
illuminated colony counter. There is also a minimum requirement of 30 colonies
(maximum to not exceed 300) for accurate results.
Results unnecessarily elaborate in cases where results are required to only
immediately confirm if food is fit for consumption.
Source : 1) ATP bioluminescence rapid detection of total viable count in soy sauce,
Luminescence, The Journal of Biological and Chemical Luminescence,14-Jun-11
2) Food Microbiology and Hygiene, By P. R. Hayes & Richard Hayes, page 189

43. Evolution of the concept of ATP
bioluminescence in food industry


Adenosine-5’-Triphosphate (or ATP) is the most important biological fuel in living
organisms, and the detection of ATP origination can be important to detect living
microorganisms such as pathogens.








LUCIFERASE

The visible glow of pathogen helps provide instant counting results.
WHAT?
HOW?
WHY?

44. Applications in the Food Industry
VALUE
PROPOSITION
Disposable
Low cost
Easy operation
Fast response

Prototype of ATP bioluminescence based Biosensor for detection of
bacteria
Source : Disposable bioluminescence-based biosensor for detection of bacterial count in food, Analytical Biochemistry 394 (2009) 1-6

45. Opportunities in the Food Industry
In a local butcher shops for detection of contaminated food substances such
as meat.

To quickly and easily detect if small individual samples (i.e. per bottle) of
fluids, such as milk or water, have been contaminated/spoiled during
packaging transportation.

In remote areas such as Saharan desert and Alaska.

Can be used by rescue workers during natural disasters such as earthquakes
and tsunamis.

Future Space missions (e.g. during a long mars mission)

46. Confectionary market growth worldwide










Source : http://www.nclcworld.com/pdf/Confectionery%20Market%20by%20Jim%20Corcoran.pdf

47. Opportunities in industry
BioLume : Bioluminescent bacteria added will be regulated as a „food additive‟ by the FDA.
Tourism opportunities in a country like Singapore.
Huge profits possible in the industry due to novelty factor.

48. Creation of dynamic art - The Center for Biofilm Engineering and the Montana
State University School of Art created the BIOGLYPHS project
Source : http://www.biofilm.montana.edu/Bioglyphs/
Opportunities for Bioluminescence in non-
traditional arenas

49. Genetically engineered Bioluminescent pets
Source : http://www.ibtimes.com/glowing-dark-rare-pictures-genetically-engineered-fluorescent-animals-photos-840089
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.

50. 51
Bioluminescence Why does Bioluminescence occur?
How it Happens ?
Bioluminescence Vs. Fluorescence Vs.
Incandescence
Opportunities Lighting
Biomedical Imaging
Food Industry
Inferences and
Conclusion
Commercialization
Entrepreneurial Opportunities
Technology Convergence
Conclusion
Outline

51. Interesting Opportunities
Plants and Trees
When crops need water or
nutrients, they'll be able to
tell farmers. Plants could
even go to red, yellow or
green "alert" to give farmers
early warning about disease
and invasions by harvest-
destroying pests.
Smart Crops Streetlights Into
Tree-lights
Replace electricity-draining
conventional streetlights, lit-
up road signs and interior
lighting. the trees would
come "on" at night and go
"off" during the day. The trees
would need only air, water,
and soil nutrients to maintain
their urban lighting duties.
Military
Biodegradable landing zone markers
Bioluminescent "friend vs. foe" identification markers and
security systems
Landing Lights
52

52. Factors for commercialization
Scalability
Maturity of technology
Entrepreneurial opportunities
Market penetration
Cost effectiveness
Need for a cheaper lighting
Lighting
Food
Contamination
BLI
Other new areas of market penetration

53

53. Lighting Opportunity
Non Nuclear Countries - need for sustainable lighting
Availability of oceans to tap Bioluminescent Bacteria
80- 85% of oceanic
world is
bioluminescent
Chemical Light
Highest efficiency
54
Legend
No power shortage
Scarcity of electricity
No huge infrastructure required or dams or gridlines

54. Food Contamination - Opportunity
Legend
National Food Center
WHO Collaborating Testing Center
Located near Oceanic
Areas
Cost involved in conventional testing is high
Time taken for the results is more
Advantage of Bioluminescent Food
contamination testing
Quicker
Cheaper
Ease of use of instrument
55
In situ – Lab to sample

55. Identifying Opportunity
Base / Environment to enable Bioluminescent
Luciferase Enzyme is the base across all three applications
Typically any bacteria can be genetically modified to produce Luciferin
Lighting Food contamination BLI - Imaging
56
Appropriate platform for sustainable culturing of Bioluminescent
organisms presents a new horizon

56. Large Scale Production Setup
Production
center for
Luciferase
Enzyme
•Can be located near areas like Food
contamination test centres /
Requirement for Bioluminescent
based lighting
•Can be used to make
Bioluminescent based lighting.
•Can employ “Made to order” lights
/ artistic pets / wall design / Bill
boards and Biosensors
•Can extract the enzyme in large
scale.
•Production of synthetic Luciferin
Research
Development
Commercialization
57

57. Technology Convergence
Nanotechnology
System on Chip
Nano particles
Nano polymers
Minimum flashing
Maximum
Luminosity
MEMS
Structural Changes
LED‟s
Bio inspired lighting
from firefly
Genetics
DNA improvements
Technology merge
Computational
DNA‟s

58. Conclusion
59
Change provides opportunities
Bioluminescence provides a new dimension to Lighting, Healthcare and Food
industry
Adoption of this technologies will lead to massive growth Bioluminescence
When technology matures and becomes economically feasible, it will definitely
offer a superior value proposition

60. Name Matric Number
Ajay Srinivasan A0102866E
Anurag Sharma A0102808M
Shuchi Dangwal A0104405X
Souvik Sen A0102840W