Bioengineering a Novel Bio-photovoltaic System

1. Add 4 µl loading dye to each sample Load 10 µl in wells 1-5 Load 10 µl marker DNA in lane 6 Run @ 80 volts for 5’, then @ 150 volts

2. Possible Projects Studying potential of Rhodopseudomonas (a.k.a. Rhodobacter) for biophotovoltaics Rosenbaum M, Schro¨ der U, Scholz F: In situ electrooxidation of photobiological hydrogen in a photobiological fuel cell based on Rhodobacter sphaeroides. Environ Sci Technol 2005, 39:6328-6333.

3. Possible Projects • 2. Studying Cyanobacteria for biophotovoltaics • Finding new ones • Tweaking knowns • http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Protein-Expression-and-Analysis/Protein-Expression/algae-engineering-kits.html

4. Possible Projects • 3. Studying algae for biophotovoltaics • especially under CO2 deprivation

5. Possible Projects • 4. Studying plants for biophotovoltaics • Roots under hypoxia • Aquatics under CO2 deprivation

6. Possible Projects 5. Engineering the production of other novel products

7. General principle: transfer e- from food to anode via direct contact, nanowires or a mediator. H+ diffuse to cathode to join e- forming H2O

8. We need to make or • acquire some electrodes! • bacteria, cyanobacteria • or green algae can coat • Uses 2-hydroxy-1,4 • -naphtoquinone as shuttle

9. We need to make or acquire some electrodes! • bacteria, cyanobacteria or green algae can coat • Uses 2-hydroxy-1,4-naphtoquinone as shuttle • b) H2- generating microbes, electrocatalytic anode • Chlamydomonas made more H2 when H2 was rapidly removed.

10. We need to make or acquire some electrodes! • bacteria, cyanobacteria or green algae can coat • Uses 2-hydroxy-1,4-naphtoquinone as shuttle • b) H2- generating cyanobacteria, electrocatalytic anode • c) Phototrophic microbes and mixed heterotrophic bacteria in sediments

11. We need to make or acquire some electrodes! • bacteria, cyanobacteria or green algae can coat • Uses 2-hydroxy-1,4-naphtoquinone as shuttle • b) H2- generating microbes, electrocatalytic anode • c) Phototrophic microbes and mixed heterotrophic bacteria in sediments • d) Plants and mixed heterotrophic bacteria in sediments • http://www.plantpower.eu/

12. Uses 2-hydroxy-1,4-naphtoquinone as shuttle • b) H2- generating microbes, electrocatalytic anode • c) Phototrophic microbes and mixed heterotrophic bacteria in sediments • d) Plants and mixed heterotrophic bacteria in sediments • e) Ex situ photosynthesis coupled with mixed • heterotrophic bacteria at a dark anode

13. Uses 2-hydroxy-1,4-naphtoquinone as shuttle • b) H2- generating microbes, electrocatalytic anode • c) Phototrophic microbes and mixed heterotrophic bacteria in sediments • d) Plants and mixed heterotrophic bacteria in sediments • e) Ex situ photosynthesis coupled with mixed • heterotrophic bacteria at a dark anode • f) Direct electron transfer between photosynthetic bacteria and electrodes

14. Uses 2-hydroxy-1,4-naphtoquinone as shuttle • b) H2- generating microbes, electrocatalytic anode • c) Phototrophs & mixed heterotrophs in sediments • d) Plants and mixed heterotrophic bacteria in sediments • e) Ex situ photosynthesis & mixed heterotrophs • f) Direct e transfer between phototrophs and electrodes • g) Use photosynthesis to generate O2 at the cathode

15. We need to make or acquire some electrodes! • bacteria, cyanobacteria or green algae can coat

16. We need to make or acquire some electrodes! • That bacteria, cyanobacteria or green algae can coat • That roots can coat

17. How to bioengineer a novel bio-photovoltaic system?

18. How to bioengineer a novel bio-photovoltaic system? Identify a suitable candidate organism or consortium

19. How to bioengineer a novel bio-photovoltaic system? • Identify a suitable candidate organism or consortium • Must be tractable to bioengineering

20. How to bioengineer a novel bio-photovoltaic system? • Identify a suitable candidate organism or consortium • Must be tractable to bioengineering • Must be able to be genetically transformed

21. How to bioengineer a novel bio-photovoltaic system? • Identify a suitable candidate organism or consortium • Must be tractable to bioengineering • Must be able to be genetically transformed • Bioengineering is changing an organism’s genotype by adding DNA encoding specific functions!

22. How to bioengineer a novel bio-photovoltaic system? • Identify a suitable candidate organism or consortium • Must be tractable to bioengineering • Must be able to be genetically transformed • Bioengineering is changing an organism’s genotype by adding DNA encoding specific functions! • Sequences must function in the new host

23. How to bioengineer a novel bio-photovoltaic system? • Identify a suitable candidate organism or consortium • Must be tractable to bioengineering • Must be able to be genetically transformed • Bioengineering is changing an organism’s genotype by adding DNA encoding specific functions! • Sequences must function in the new host • Must contain binding sites for the host regulatory proteins

24. How to bioengineer a novel bio-photovoltaic system? • Identify a suitable candidate organism or consortium • Must be tractable to bioengineering • Must be able to be genetically transformed • Bioengineering is changing an organism’s genotype by adding DNA encoding specific functions! • 2. Sequences must function in the new host • Must be replicated

25. How to bioengineer a novel bio-photovoltaic system? • 2. Sequences must function in the new host • Must be replicated • Either insert into chromosome

26. How to bioengineer a novel bio-photovoltaic system? • 2. Sequences must function in the new host • Must be replicated • Either insert into chromosome, or replicate extrachromasomally (i.e. as a plasmid)

27. How to bioengineer a novel bio-photovoltaic system? • 2. Sequences must function in the new host • Must be replicated • Either insert into chromosome, or replicate extrachromasomally (i.e. as a plasmid) • Must have origin of replication that functions in new host (highly species-specific in bacteria)