2012 iGEM - University of Lethbridge

Project Overview

Increasing global oil demands require new, innovative technologies for the extraction of unconventional oil sources such as those found in Alberta’s Carbonate Triangle. Carbonate oil deposits account for almost 50% of the world’s oil reserves and approximately 26% of the bitumen found in Alberta 1. Due to unstable oil prices in Western Canada, these vast reserves have historically been set aside in favour of less time consuming, more economical sites. Microbial enhanced oil recovery (MEOR) has been utilized across the world to increase the productivity of difficult resources including carbonate oil deposits. Using a synthetic biology approach, we have designed the CAB (CO2, acetic acid, and biosurfactant) extraction method that demonstrates a modified MEOR method for extracting carbonate oil deposits. CAB extraction will utilize the natural carbon fixation machinery in the cyanobacteria Synechococcus elongatus to convert CO2 into sugars to fuel acetic acid and biosurfactant production in Escherichia coli. Acetic acid applied to carbonate rock increases the pore sizes and allows for enhanced oil recovery. The reaction produces gases that will help pressurize the well site to facilitate extraction. The natural biosurfactant rhamnolipid will also be applied to the carbonate rock to further enhance extraction yields.

By coupling carbon capture with acetic acid and biosurfactant production, carbonate oil deposits can be mined with reduced greenhouse gas emissions. The use of carbon fixation to feed downstream systems can be tailored for use as a module in many applications requiring inexpensive methods for fueling biological systems. CAB extraction will be suitable for large-scale bioreactors, providing an alternative, inexpensive, and environmentally sustainable method for MEOR from Alberta’s oil deposits. Furthermore, developing the carbon capture module will be of interest in oil extraction strategies using steam, as it will help with the mitigation of CO2 release caused by steam production using for example natural gas.


1. Hryhor, D. W. Emerging Solutions for Heavy Oil Production from Carbonates. (TAMM Oil and Gas Corp., 2008).

2. Bott, R. D. Canada's Oil Sands. (Canadian Centre for Energy Information, 2011).

3. Buijse, M., de Boer, P., Breukel, B. & Burgos, G. Organic acids in carbonate addizing. Spe Production & Facilities 19, 128-134 (2004).

4. World Energy Outlook 2010 - Executive Summary. (International Energy Agency, 2010).

5. MicroPro-Gmbh. MEOR case study - oil field “Doebern” (Germany), (2011).

6. Sen, R. Biotechnology in petroleum recovery: The microbial EOR. Progress in Energy and Combustion Science 34, 714-724, doi:10.1016/j.pecs.2008.05.001 (2008).

7. Niederholtmeyer, H., Wolfstaedter, B. T., Savage, D. F., Silver, P. A. & Way, J. C. Engineering Cyanobacteria To Synthesize and Export Hydrophilic Products. Applied and Environmental Microbiology 76, 3462-3466, doi:10.1128/aem.00202-10 (2010).

8. Iancu, C. V. et al. Organization, Structure, and Assembly of α-Carboxysomes Determined by Electron Cryotomography of Intact Cells. Journal of Molecular Biology 396, 105-117, doi:10.1016/j.jmb.2009.11.019 (2010).

9. Bonacci, W. et al. Modularity of a Carbon-Fixing Protein Organelle. Proceedings of the National Academy of Sciences of the United States of America 109, 478-483, doi:10.1073/pnas.1108557109 (2012).

10. Price, G. D. Inorganic carbon transporters of the cyanobacterial CO2 concentrating mechanism. Photosynthesis Research 109, 47-57, doi:10.1007/s11120-010-9608-y (2011).

11. Bar-Even, A., Noor, E., Lewis, N. E. & Milo, R. Design and Analysis of Synthetic Carbon Fixation Pathways. Proceedings of the National Academy of Sciences 107, 8889-8894, doi:10.1073/pnas.0907176107 (2010).

12. Davies, S. & Kelkar, S. Carbonate Stimulation. Middle East & Asia Reservoir Review 8, 52-63 (2007).

13. Ferry, J. G. in Methods in Enzymology: Methods in Methane Metabolism, Pt A Vol. 494 Methods in Enzymology (eds A. C. Rosenzweig & S. W. Ragsdale) 219-231 (Elsevier Academic Press Inc, 2011).

14. Latimer, M. T. & Ferry, J. G. Cloning, Sequence-Analysis, and Hyperexpression of the Genes Encoding Phosphotransacetylase and Acetate Kinase from Methanosarcina thermophila. J. Bacteriol. 175, 6822-6829 (1993).

15. Nakano, S., Fukaya, M. & Horinouchi, S. Putative ABC Transporter Responsible for Acetic Acid Resistance in Acetobacter aceti. Applied and Environmental Microbiology 72, 497-505, doi:10.1128/aem.72.1.497-505.2006 (2006).

16. Wang, Q. et al. Engineering Bacteria for Production of Rhamnolipid as an Agent for Enhanced Oil Recovery. Biotechnology and Bioengineering 98, 842-853, doi:10.1002/bit.21462 (2007).

17. Patel, R. M. & Desai, A. J. Biosurfactant production by Pseudomonas aeruginosa GS3 from molasses. Letters in Applied Microbiology 25, 91-94, doi:10.1046/j.1472-765X.1997.00172.x (1997).

18. iGEM2011. Results Summary, (2011).

19. Delebecque, C. J., Lindner, A. B., Silver, P. A. & Aldaye, F. A. Organization of Intracellular Reactions with Rationally Designed RNA Assemblies. Science 333, 470-474, doi:10.1126/science.1206938 (2011).

20. Ohno, H. et al. Synthetic RNA-protein complex shaped like an equilateral triangle. Nature Nanotechnology 6, 115-119, doi:10.1038/nnano.2010.268 (2011).

21. Zhu, C. & Ye, Q. [Selection of acetate-tolerant mutants from Escherichia coli DH5alpha and the metabolic properties of mutant DA19]. Wei Sheng Wu Xue Bao 43, 460-465 (2003).

22. Ochsner, U. A., Fiechter, A. & Reiser, J. Isolation, Characterization, and Expression in Escherichia coli of the Pseudomonas aeruginosa rhlAB Genes Encoding a Rhamnosyl Transferase Involved in Rhamnolipid Biosurfactant Synthesis. Journal of Biological Chemistry 269, 19787-19795 (1994).

23. iGEM2011. BioBricks, (2011).

24. Ohlendorf, R., Vidavski, R. R., Eldar, A., Moffat, K. & Möglich, A. From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression. Journal of Molecular Biology 416, 534-542, doi:10.1016/j.jmb.2012.01.001 (2012).

25. Flynn, J. M., Neher, S. B., Kim, Y.-I., Sauer, R. T. & Baker, T. A. Proteomic Discovery of Cellular Substrates of the ClpXP Protease Reveals Five Classes of ClpX-Recognition Signals. Molecular Cell 11, 671-683, doi:10.1016/s1097-2765(03)00060-1 (2003).