Potential Algal Biofuel Applications in West Virginia STa
Potential Algal Biofuel Applications in West Virginia STa. R Symposium October 23, 2013 Derrick R. J. Kolling Department of Chemistry Derrick R. J. Kolling March 16, 2010
• Biofuels overview Talk Overview – Why do we need biofuels? • The light reactions of photosynthesis 2 H 2 O + 4 h 4 H+ + 4 e- + O 2 • Tools of the trade – – O 2 electrode Fluorometer EPR spectrometer GC-MS • Research projects – – OEC photoassembly Temperature dependence of molecular photosynthesis Algal lipidomics Biofuels from invasive algal species Derrick R. J. Kolling October 23, 2013
Renewable Energy Biofuels Solar power Wind power Tidal power Hydropower Geothermal futurefarmers. com/survey/algae epmb. berkeley. edu
Dismukes et al. (2008) Derrick R. J. Kolling October 23, 2013
Derrick R. J. Kolling October 23, 2013
lipid protein Biomass biodiesel animal feed carbohydrate CO 2 anaerobic Carbon Fixation Protein Synthesis O 2 Lipid Synthesis Fermentation ATP Electrons (as NADH, NADPH, Reduced Ferredoxin) CO 2, organic acids e- ATP Photosynthesis H 2 O H 2 ethanol O 2 Source: Damian Carrieri Derrick R. J. Kolling March 16, 2010
Light Reactions Dismantling photosynthesis from the organism to the molecules…. Derrick R. J. Kolling October 23, 2013
Light Reactions …from the molecular to the atomic level Barber & Iwata (2004) Science Umena et al. (2011) Nature Derrick R. J. Kolling October 23, 2013
Research Tools • Oximetry • GC-MS • Fluorescence • Electron Paramagnetic Resonance Derrick R. J. Kolling October 23, 2013
OEC Photoassembly O 2 evolution rate (µmol O 2/mg Chl a/h) Temperature dependence of oxygen evolution 900 800 700 600 500 400 300 200 100 0 BBY particles w/ out PSII DCBQ BBY particles w/DCBQ PSII w/DCBQ Normalized BBYs w/out PSII DCBQ normalized 0 20 40 Temperature °C 60
OEC Photoassembly Temperature dependence of oxygen evolution vs. photoassembly Intact PSII Photoassembled PSII at 28°C -Share a rate-limiting step
OEC Photoassembly Percent Recovery Oxygen Evolution Percent Recovery 120 100 80 60 40 20 0 5 10 15 20 22 25 26 27 28 29 30 32 35 40 Temperature °C -Protease activity explains dip
OEC Photoassembly Findings • Oxygen evolution and OEC photoassembly share a 28°C maximum • OEC photoassembly and PSII oxygen evolution appear to share a rate-limiting step • Thermal inactivation of PSII and apo-PSII occurs at temperatures greater than 28°C • Thermal inactivation is partially due to protease activity (Deg and/or Fts. H? )
OEC Photoassembly Application Bioinspired/biomimicked System www. ruhr-uni-bochum. de/h 2 design/profile/main. html Artificial Leaf www 3. imperial. ac. uk
Algal Lipidomics Comparison of lipid accumulation in photomixotrophically and heterotrophically grown Chlorella vulgaris cultures
Algal Lipidomics 4 5 6 7 [ [ 3 [ [ [ 2 [ 1 [ 0 [ [ 1 2 Culture 3 4 5 6 Measured: -[Chl a] -turbidity -dry weight -lipid dry weight 8
Algal Lipidomics Chl a Expression Cell Density 750 nm 35 30 30 25 25 Chl a (mg/ml) Turbidity (AU) 20 15 10 Photomixotrophic Heterotrophic 5 0 0 0 1 2 3 4 5 Time (Days 6 7 8 9 0 1 2 3 4 5 6 Time (Days) Derrick R. J. Kolling 7 8 9 October 23, 2013
Algal Lipidomics Dry Weight Growth Curves and Corresponding Dry Lipid Weights Weight (mg) 10 8 Photomixotrophic Lipids Seri es 1 6 Heterotrophic Lipids Seri es 2 Photomixotrophic 4 Seri es 3 Heterotrophic Cells 2 0 0 1 2 3 4 5 Time (Days) C 16: palmitic acid C 18: stearic acid C 18: 1(n-9): oleic acid C 18: 2(n-6): linoleic acid 6 7 8 9
Algal Lipidomics Findings • Photomixotrophically grown cells produce 2 X as many lipids as do heterotrophically grown cells • Photomixotrophically grown cells reach stationary phase and higher biomass sooner • Cells produce palmitic, stearic, oleic, and linoleic acids
Algal Lipidomics Application Chisti (2007) Biotech. Advances
Acknowledgements • Temperature Dependence of PSII -James Board, Hope Cook, Matt Thompson, Shane Kagen, Jordan Hilgeford, Justin Erwin • OEC photoassembly -James Board, Hope Cook, Ben Blodgett, Matt Thompson, Shane Kagen, Jordan Hilgeford, Chase Turner, Ben Weiner • Algal lipidomics A c k n o w l e d g e m e n t s -Ben Woodworth, Tony Stephenson, Rebecca Mead, Courtney Nichols, Prof. Leslie Frost (MU) • Bioethanol from invasive algal species Funding -Kevin Militello, Shaheed Elhamdani This material is based upon work supported by the National Science Foundation under Cooperative Agreement Award number EPS-1003907 and CHE 1229498. Derrick R. J. Kolling October 23, 2013
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