E. cowli. Fighting Climate Change at the Source

Transcription

1 E. cowli Fighting Climate Change at the Source

2 The Greenhouse Effect Promotes Global Warming igem Team Braunschweig 2

3 Global Temperature Keeps Rising Source: NASA igem Team Braunschweig 3

4 Why We Target Methane Greenhouse Gases Global Warming Potential Others Methane 8% 20% 72% Carbon dioxide Methane has a 25 times greater Global Warming Potential than carbon dioxide! igem Team Braunschweig Source: United States Environmental Protection Agency 4

5 Cattle the Unintended Climate Killers Annual methane emissions igem Team Braunschweig Source: NASA s Goddard Institute for Space Science 5

6 Spread the Word! To beef or not to beef? igem Team Braunschweig 6

7 To Beef or not to Beef? Would you change your dietary behavior for the sake of the climate? Other No Yes igem Team Braunschweig 7

8 Previous Approaches Targeted the Microbiota Hydrogen Methane Protozoa Methanogens igem Team Braunschweig 8

9 Our Approach Targets Methane Hydrogen Methane Protozoa Methanogens E. cowli igem Team Braunschweig 9

10 We Fight Global Warming at the Source igem Team Braunschweig 10

11 Problems We Have to Face Activity Production Application Ingredient Feasibility igem Team Braunschweig 11

12 Methylococcus capsulatus Grows too Slowly doubling times: E. coli: 20 min x 120 M. capsulatus: 40 h [1] growth curve of Methylococcus capsulatus igem Team Braunschweig [1] Khalifa

13 Methane Monooxygenase - our Secret Ingredient CH 4 + O 2 + NADH + H + CH 3 OH + H 2 O + NAD + soluble methane monooxygenase (smmo) Reductase Hydroxylase igem Team Braunschweig 13

14 Methane Monooxygenase - our Secret Ingredient CH 4 + O 2 + NADH + H + CH 3 OH + H 2 O + NAD + soluble methane monooxygenase (smmo) Reductase Hydroxylase igem Team Braunschweig 14

15 Boston, We Have a Folding Problem! igem Team Braunschweig 15

16 Addition of Chaperones Allows Soluble Expression Coexpression with chaperones (GroEL, GroES) led to soluble expression of the remaining subunits! igem Team Braunschweig 16

17 Addition of Chaperones Allows Soluble Expression Coexpression with chaperones (GroEL, GroES) led to soluble expression of the remaining subunits! igem Team Braunschweig 17

18 E. Cowli Was Born! E. coli E. cowli soluble methane monooxygenase igem Team Braunschweig 18

19 E. cowli Degrades Methane! 2% E. coli E. cowli The subunits form an active complex E. cowli works! igem Team Braunschweig 19

20 E. cowli Degrades Methane! 2% E. coli E. cowli The subunits form an active complex E. cowli works! igem Team Braunschweig 20

21 Can E. cowli Survive under Rumen Conditions? Source: Hoards Dairyman igem Team Braunschweig 21

22 E. cowli Does not Grow on Rumen Fluid Glucose + Glucose agar plates with 100% sterile filtrated rumen fluid inoculated with unfiltrated rumen fluid E. cowli unfiltrated rumen fluid E. cowli igem Team Braunschweig 22

23 Alginate Beads Can Be Used to Protect E. cowli E. cowli is able to grow inside alginate beads. The ingredients of the beads are in no way toxic for E. cowli or the cow. Dr. Ulf Prüße, Thünen Institute Braunschweig igem Team Braunschweig 23

24 E. cowli Works in Rumen Fluid 2% igem Team Braunschweig 24

25 E. cowli Works in Rumen Fluid 2% igem Team Braunschweig 25

26 Cow Farts Are Dangerous! Source: University of Auckland, New Zealand igem Team Braunschweig 26

27 The Model Matches the Measurements 2% igem Team Braunschweig 27

28 750 Beads for a Methane-Free Cow igem Team Braunschweig 28

29 50 $ per Year for a Better Climate 110 kg 750 x igem Team Braunschweig 29

30 50 $ per Year for a Better Climate 50 US$ correspond to 6% of the running costs for keeping a cow! 110 kg 50 US$ igem Team Braunschweig 30

31 50 $ per Year for a Better Climate 164 million tonnes igem Team Braunschweig 31

32 50 $ per Year for a Better Climate 164 million tonnes igem Team Braunschweig 32

33 50 $ per Year for a Better Climate igem Team Braunschweig 33

34 Our Achievements! E. cowli engineered! E. cowli works! Application approved! Ingredient isolated! Feasibility shown! igem Team Braunschweig 34

35 13 BioBricks Submitted! 10 new BioBricks created 2 BioBricks improved 1 Protein Generator created: complete soluble methane monooxygenase, easily controllable, methane-degrading igem Team Braunschweig 35

36 Our Thanks Go to Prof. Dr. Stefan Dübel Dr. Torsten Meyer Tobias Unkauf and Jonas Zantow Institute of Biochemistry, Biotechnology and Bioinformatics Technische Universität Braunschweig Thünen Institute Braunschweig for technical assistance igem Team Aachen for providing us with a methane sensor and all our other supporters! igem Team Braunschweig 36

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38 Our Intern Benjamin These two weeks were eventful and exciting for me I got a great insight into biotechnology, especially the lab work. igem Team Braunschweig 38

39 Media igem Team Braunschweig 39

40 Sources of Methane Emissions igem Team Braunschweig 40

41 Safety Kill Switch igem Team Peking 2012 igem Team Braunschweig 41

42 Global Warming Potential igem Team Braunschweig 42

43 Interlab Study I2020 GFP construct 0 K E0240 GFP construct I K E0240 GFP construct II K K in psb3k3 RFP construct 0 K K RFP construct I K K RFP construct II igem Team Braunschweig 43

44 MQ-4 Methane Sensor igem Team Braunschweig 44