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E. cowli
Fighting Climate Change at the Source
The Greenhouse Effect Promotes Global Warming
2iGEM Team Braunschweig
Global Temperature Keeps Rising
iGEM Team Braunschweig 3
Source: NASA
Why We Target Methane
iGEM Team Braunschweig 4Source: United States Environmental Protection Agency
Methane has a 25 times greater Global Warming Potential than carbon dioxide!
Carbon dioxide
Methane
Others
Greenhouse Gases Global Warming Potential
72%20%
8%
Cattle – the Unintended Climate Killers
iGEM Team Braunschweig 5Source: NASA‘s Goddard Institute for Space Science
Annual methane emissions
Spread the Word!
iGEM Team Braunschweig 6
To beef ornot to beef?
To Beef or not to Beef?
iGEM Team Braunschweig 7
Would you change
your dietary behavior for the sake of the climate?
No
Other
Yes
Previous Approaches Targeted the Microbiota
iGEM Team Braunschweig 8
Hydrogen Methane
Protozoa Methanogens
iGEM Team Braunschweig 9
Hydrogen Methane
Protozoa MethanogensE. cowli
Our Approach Targets Methane
We Fight Global Warming at the Source
iGEM Team Braunschweig 10
Problems We Have to Face
iGEM Team Braunschweig 11
Ingredient
Production
Activity
Application
Feasibility
Methylococcus capsulatus Grows too Slowly
doubling times:
E. coli: 20 min
M. capsulatus: 40 h[1]
iGEM Team Braunschweig 12[1] Khalifa 2013
growth curve of Methylococcus capsulatus
x 120
Reductasesoluble methanemonooxygenase
(sMMO)
Methane Monooxygenase - our Secret Ingredient
CH4 + O2 + NADH + H+ → CH3OH + H2O + NAD+
iGEM Team Braunschweig 13
Hydroxylase
Reductasesoluble methanemonooxygenase
(sMMO)
Methane Monooxygenase - our Secret Ingredient
CH4 + O2 + NADH + H+ → CH3OH + H2O + NAD+
iGEM Team Braunschweig 14
Hydroxylase
Boston, We Have a Folding Problem!
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Addition of Chaperones Allows Soluble Expression
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Coexpression with chaperones (GroEL, GroES) led to soluble expression of the remaining subunits!
Addition of Chaperones Allows Soluble Expression
iGEM Team Braunschweig 17
Coexpression with chaperones (GroEL, GroES) led to soluble expression of the remaining subunits!
E. Cowli Was Born!
iGEM Team Braunschweig 18
E. coli E. cowlisoluble methanemonooxygenase
E. cowli Degrades Methane!
iGEM Team Braunschweig 19
E. coli
E. cowli
The subunits form an active complex – E. cowli works!
2%
E. cowli Degrades Methane!
iGEM Team Braunschweig 20
E. coli
E. cowli
The subunits form an active complex – E. cowli works!
2%
Can E. cowli Survive under Rumen Conditions?
iGEM Team Braunschweig 21
Source: Hoards Dairyman
E. cowli Does not Grow on Rumen Fluid
iGEM Team Braunschweig 22
unfiltratedrumen fluid
E. cowliunfiltratedrumen fluid
E. cowli
+ Glucose – Glucose
inoculated with
agar plateswith 100%
sterile filtratedrumen fluid
Alginate Beads Can Be Used to Protect E. cowli
iGEM Team Braunschweig 23
„E. cowli is able to grow inside alginatebeads. The ingredients of the beads arein no way toxic for E. cowli or the cow.“
Dr. Ulf Prüße, Thünen Institute Braunschweig
E. cowli Works in Rumen Fluid
iGEM Team Braunschweig 24
2%
E. cowli Works in Rumen Fluid
iGEM Team Braunschweig 25
2%
Cow Farts Are Dangerous!
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Source: University of Auckland, New Zealand
The Model Matches the Measurements
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2%
750 Beads for a Methane-Free Cow
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50 $ per Year for a Better Climate
iGEM Team Braunschweig 29
750 x
110 kg
50 $ per Year for a Better Climate
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50 US$ correspond to 6% of the running costs for keeping a cow!
50 US$
110 kg
50 $ per Year for a Better Climate
iGEM Team Braunschweig 31
164 milliontonnes
50 $ per Year for a Better Climate
iGEM Team Braunschweig 32
164 milliontonnes
50 $ per Year for a Better Climate
iGEM Team Braunschweig 33
Our Achievements!
iGEM Team Braunschweig 34
Ingredientisolated!
E. cowliengineered!
E. cowliworks!
Applicationapproved!
Feasibilityshown!
13 BioBricks Submitted!
• 10 new BioBricks created
• 2 BioBricks improved
• 1 Protein Generator created:
complete soluble methanemonooxygenase,
easily controllable,
methane-degrading
iGEM Team Braunschweig 35
Our Thanks Go to …
iGEM Team Braunschweig 36
Prof. Dr. Stefan DübelDr. Torsten Meyer
Tobias Unkauf and Jonas ZantowInstitute of Biochemistry, Biotechnology and Bioinformatics
Technische Universität Braunschweig
Thünen Institute Braunschweigfor technical assistance
iGEM Team Aachenfor providing us with a methane sensor
and all our other supporters!
37
Our Intern Benjamin
iGEM Team Braunschweig 38
“These two weeks were eventful and exciting for me … I got a great insight into biotechnology,
especially the lab work.“
Media
iGEM Team Braunschweig 39
Sources of Methane Emissions
iGEM Team Braunschweig 40
Safety – Kill Switch
iGEM Team Braunschweig 41
© iGEM Team Peking 2012
Global Warming Potential
iGEM Team Braunschweig 42http://www.co2crc.com.au/
Interlab Study
iGEM Team Braunschweig 43
I2020 GFP construct 0K823005 + E0240 GFP construct IK823012 + E0240 GFP construct II
K823005 + K516032 in pSB3K3 RFP construct 0K823005 + K516032 RFP construct IK823012 + K516032 RFP construct II
MQ-4 Methane Sensor
iGEM Team Braunschweig 44
