2012 Advisory Panel The Challenge of Algal Fuel: Monetizing the Entire Algal Biomass Rhykka Connelly Center for Electromechanics The University of Texas at Austin 12/4/2012 Timeline of the Biomass Program Sugar Cane 2005 2007 2009 2012 2014 Sugar Cane Production • The extraction rate should be at ≥ 9% to reach ethanol producing targets • In reality, the conventional extraction rate ranges only between 5-6% • Need extraction to improve by 33-45% The CEM Solution Marx Bank High Voltage Generator The CEM Solution Brix values of extracted juice (100 pulses) 1.2 Average Brix difference: 0.37 1.0 Brix 0.8 Test 0.6 Control 0.4 0.2 0.0 A B • Electroporation of sugar cane cell walls • Extraction improved by 37% on average C Avg Algae Sugar Cane 2005 Algae 2007 Algae Algae Chlorella sp. lipid bodies Algae Structure and Products The CEM Solution: Breaking Algae with Electric Fields • Convenient, adjustable voltage source • Does not provide optimum wave shape • Not efficient or practical for field use The CEM Solution: Multidisciplinary Approach • Algae Group – ≈50 faculty, researchers, and students – Plus larger group of researchers in associated, related fields • Multidisciplinary – Biologists, biochemists, physicists, mechanical engineers, electrical engineers, chemical engineers, and environmental engineers • Focused on Fuel – But Working on Cash Flow from CoProducts along the Way – Complete Process • • • • • • • • Algae selection – key driver of economics Growth—as needed Harvesting Dewatering Lysing Separation Metrology – without good process measurements, there is no process control Life cycle analysis – program focus, regulatory acceptance Algae Processing Options concentrate algae break algae product recovery lipids/oils for fuel centrifuge solvent extraction centrifugation drum drier pH-driven flocculation expeller press hollow fiber membrane carotenoids chemical flocculation electric field sonication direct transesterification carbohydrates bioflocculation dissolved air flotation supercritical fluid mixotrophic growth bead-milling solvent phase partitioning omega 3-6-9 fatty acids clean biomass for fertilizer/animal feed Algae Processing Options concentrate algae break algae product recovery lipids/oils for fuel centrifuge solvent extraction centrifugation drum drier pH-driven flocculation expeller press hollow fiber membrane carotenoids chemical flocculation electric field sonication direct transesterification carbohydrates bioflocculation dissolved air flotation supercritical fluid mixotrophic growth bead-milling solvent phase partitioning omega 3-6-9 fatty acids clean biomass for fertilizer/animal feed The CEM Solution concentrate algae break algae product recovery lipids/oils for fuel centrifuge solvent extraction centrifugation drum drier pH-driven flocculation expeller press hollow fiber membrane carotenoids chemical flocculation electric field sonication direct transesterification carbohydrates bioflocculation dissolved air flotation supercritical fluid mixotrophic growth bead-milling solvent phase partitioning omega 3-6-9 fatty acids clean biomass for fertilizer/animal feed Algae Processing Development Solid State Lysing Power Supply • Patents filed • Paper* published * S. Bae, A. Kwasinski, M. M. Flynn, and R. E. Hebner, “High-Power Pulse Generator with Flexible Output Pattern,” IEEE Trans. Power Electronics, 25(7): 1675-1684. Algae Processing Development: Creating Analytical Tools for the Industry Nile Red • Industry standard for measuring lipids in total Identification of lipids by NMR Simultaneous detection of lipids by ELSD and mass spectrometer • Non-specific for non-polar lipids (hydrocarbons, some carotenoids, triacyglycerols, diglycerides, some lipoproteins), thus cannot render an accurate profile of lipids to determine downstream processing C. M. Beal, M. E. Webber, R. S. Ruoff, and R. E. Hebner, “Lipid Analysis of Neochloris oleoabundans by Liquid State NMR,” 2010, Biotechnology and Bioengineering, 106:4. Jones, J., Manning, S., Montoya, M., Keller, K., & Poenie, M. Extraction of algal lipids and their analysis by HPLC and mass spectrometry. Journal of the American Oil Chemists' Society, 2012, 89, 1371– 1381. Algae Processing Development: Creating Analytical Tools for the Industry Mass and Energy Balance Processing Flowchart Cultivation Batch Record (AC) CEM Harvest: Batch Record (AH) EWRE effluent recycled back to pond * Beal CM, Hebner RE, Romanovicz DK, Connelly RL. “Progression of lipid profile and cell structure in a research-scale production pathway for algal biocrude,” Renewable Energy. 2012, 50:86-93. Samples are collected before and after each processing step. Volume, biomass, and lipid content are measured. Cellular morphologies are monitored. Lysing: Batch Record (AL) CEM * Beal C, Smith C, Webber M, Ruoff R, Hebner R (2011) A framework to report the production of renewable diesel from algae. BioEnergy Res 4:36–60 * Beal CM, Stillwell AS, King CW, Cohen S, Berberoglu H, Bhattarai RP; Connelly RL, Webber ME, Hebner RE. “Energy return on investment for algal biofuel production coupled with wastewater treatment,” Water Environment Research. 2012, 84(9):692-710. Extraction: Batch Record (AE) SRP final biomass final oil CEM Algae Processing Technologies concentration/dewatering unit cell breaking unit oil/product recovery unit Growth and Processing Analysis via UT Technologies C.M. Beal, R.E. Hebner, M.E. Webber, R.S. Ruoff, A.F. Seibert. The energy return on investment for algal biocrude: results for a research production facility. BioEnergy Research. 2011, pp. 1–22 Transfer of CEM Algae Processing Technologies to Industry Assembled Mobile Biomass Processing Unit Algae Program Publications C.M. Beal, R.E. Hebner, M.E. Webber, “Thermodynamic analysis of algal biocrude production,” Energy. 2012, 44(1):925-943. C.M. Beal, M.E. Webber, R.S. Ruoff, R.E. Hebner, “Lipid Analysis of Neochloris oleoabundans by Liquid State NMR,” Biotechnology and Bioengineering, 2010, 106(4). S. Bae, A. Kwasinski, M.M. Flynn, R.E. Hebner, “High-Power Pulse Generator with Flexible Output Pattern,” IEEE Trans. Power Electronics, 2010, 25(7):1675-1684. C.M. Beal, R.E. Hebner, D.K. Romanovicz, R.L. Connelly, “Progression of lipid profile and cell structure in a research-scale production pathway for algal biocrude,” Renewable Energy. 2012, 50:86-93. C. Beal, C. Smith, M. Webber, R. Ruoff, R. Hebner, “A framework to report the production of renewable diesel from algae,” BioEnergy Res. 2011, 4:36–60. C.M. Beal, A.S. Stillwell, C.W. King, S. Cohen, H. Berberoglu, R.P. Bhattarai, R.L. Connelly, M.E. Webber, R.E. Hebner, “Energy return on investment for algal biofuel production coupled with wastewater treatment,” Water Environment Research. 2012, 84(9):692-710. R.V. Pearsall, R.L. Connelly, M.E. Fountain, C.S. Hearn, M.D. Werst, R.E. Hebner, E.F. Kelley EF, “Electrically dewatering microalgae,” IEEE Trans. Dielectrics and Electrical Insulation, 2011, 18(5): 1578-1583. J. Jones, C.-H. Lee, J. Wang, M. Poenie, “Use of anion exchange resins for one-step processing of algae from harvest to biofuel,” Energies, 2012, 5(7):2608-2625. J. Jones, S. Manning, M. Montoya, K. Keller, M. Poenie, ”Extraction of algal lipids and their analysis by HPLC and mass spectrometry,” Journal of the American Oil Chemists' Society, 2012, 89, 1371–1381. A. Ozkan, K. Kinney, H. Berberoglu, “Reduction of water and energy requirement of algae cultivation using an algae biofilm photobioreactor,” Bioresour. Technol., 2012, 114:542-548. Algae Program Publications A. Wileman, A. Ozkan, H. Berberoglu, “Rheological properties of algae slurries for minimizing harvesting energy requirements in biofuel production,” Bioresour. Technol., 104 (2012), pp. 432-439. H. Berberoglu, J. Jay, L. Pilon, “Effect of nutrient media on photobiological hydrogen production by Anabaena variabilis ATCC 29413,” Int. J. Hydrogen Energy, 33 (2008), pp. 1172-1184. Murphy T., Berberoglu H., “Temperature fluctuation and evaporative loss Rate in an algae biofilm photobioreactor,” ASME J. Solar Energy Engineering, 2012, vol. 134, pp. 011002-1-9. Murphy T., Berberoglu H., “Effect of algae pigmentation on photobioreactor productivity and scale-up: a light transfer perspective,” J. Quantitative Spectroscopy and Radiative Transfer, 2011, vol.112, pp. 2826-2834. Berberoglu H., Pilon L., “Maximizing the solar to H2 energy conversion efficiency of outdoor photobioreactors using mixed cultures,” International J. of Hydrogen Energy, 2010, vol. 35, pp. 500-510. Berberoglu H., Gomez P., Pilon L., 2009, “Radiation characteristics of Botryococcus braunii, Chorococcum littorale and Chlorella sp. used for CO2 fixation and biofuel production, J. Quantitative Spectroscopy and Radiative Transfer, vol. 110, pp. 1879-1893. Berberoglu H., Pilon L., Melis A., 2008, “Radiation characteristics of Chlamydomonas reinhardtii and its truncated chlorophyll antenna transformants,” International J. Hydrogen Energy, vol. 33, pp 6467-6483. Berberoglu H., Pilon L., Jay J., 2008, “Effect of nutrient media on photobiological hydrogen production of Anabaena variabilis ATCC 29413,” International J. Hydrogen Energy, vol. 33, pp. 1172-1184. Berberoglu H., Barra N., Jay J., Pilon L., 2008, “Growth, CO2 consumption H2 production of Anabaena variabilis ATCC 29413-U under different irradiances and CO2 concentrations,” J. Applied Microbiology, vol. 104, pp. 105-121. Algae Program Intellectual Property Ozkan A, Kinney K, Katz, K. Novel, “Algae biofilm photobiotreactor for reduced energy and water usage,” Energy Systems Analysis. 2010, 5:75-80. R.L. Connelly, R.V. Pearsall, R. Enhkbaatar, “Algal extracts as a wound healing agent,” UTID 5935-CON disclosure submitted Oct. 2012. P. Kipp, R.L. Connelly, F. Siebert, “Non-dispersive process for insoluble oil recovery from aqueous solutions,” submitted Jan. 2012, 20120184759. A.F. Seibert, M. Poenie, P. Kipp, R.L. Connelly, R.E. Hebner, M.D. Werst, L.E. Katz, K.A. Kinney, “Continuous oil recovery from growth environments,” UTID 5623-SEI CIP submitted Oct. 2011, provisional 61/006,342, filed Nov. 2011. P. Kipp, R.L. Connelly, F. Siebert, M. Poenie, R.E. Hebner, M.D. Werst, L.E. Katz, K.A. Kinney, “Non-dispersive process for insoluble oil recovery from aqueous solutions,” submitted Oct. 2011, 20120208247. R.E. Hebner, K.R. Davey, M.D. Werst, R.L. Connelly, “Electromechanical lysing of algae cells,” submitted July 2011, 20120021481. R.E. Hebner, M.D. Werst, A.F. Seibert, L.E. Katz, K.A. Kinney, M. Poenie, C.M. Beal, R.L. Connelly, “On-line test equipment and analysis procedure to determine material balance in algal processing for biofuel production,” provisional 61/471,597. R.L. Connelly, R.V. Pearsall, P. Enhkbaatar, B.J. Morrison, R.E. Hebner, M.D. Werst, A.F. Seibert, L.E. Katz, K.A. Kinney, M. Poenie, “Organic fertilizer derived from processed algal biomass,” UTID 5936-CON submitted Jan. 2011. R.L. Connelly, R.V. Pearsall, D. Schmid, “A method to extract and stably maintain carotenoids from algae, fungi, or bacteria,” UTID 5910-CON submitted Nov. 2010. R.E. Hebner, M.D. Werst, A.F. Seibert, L.E. Katz, K.A. Kinney, M. Poenie, R.L. Connelly, R.V. Pearsall, T.A. Beets, “A continuous process to concentrate, lyse, and extract chemicals of interest, including omega fatty acids, from algae or cyanobacteria grown phototrophically or heterotrophically,” UTID 5886-HEB submitted Sept. 2010. R.L. Connelly, R.V. Pearsall, “TLC lipid quantification kit,” UTID 5835-CON submitted Aug. 2010. Focus for the Future – Leveraging UT Expertise and Resources Sugar Cane 2005 Algae 2007 Education and Development of Industry Standards 2012 Focus for the Future – Leveraging UT Expertise and Resources Sugar Cane 2005 Algae 2007 Education and Services 2012 - 2013 • Establishment of the UT-Science and Technology Facility • Monthly Industrial Recovery & Analysis of Biomass Oils Workshops • In collaboration with the UTEX Algae Culture Collection Workshop series • students to start-ups to established companies • Analytical Service Center • Provide innovative analytical support to the industry Educating the Algae Industry Industrial Recovery & Analysis of Biomass Oils Workshop • Tour of the UT-ASTF facility • Presentations detailing current and cutting-edge processing technologies • Biochemistry of algae and popular and cutting-edge lipid analysis techniques • Hands-on laboratory instruction to extract algal products and perform detailing analyses Providing Innovative Analytical Services to the Algae Industry UT-ASTF ANALYTICAL SERVICE CENTER • LIPID EXTRACTION • HPLC/MS AND GC/MS LIPID PROFILING • SPECTROSCOPY WEBSITE: algaebiofuels.utexas.org EMAIL: r.connelly@cem.utexas.edu OFFICE: 512.232.1633 • HIGH RESOLUTION MICROSCOPY • R & D SERVICES TO OPTIMIZE GROWTH AND PROCESSING RELATIVE TO TARGET LIPIDS 1 UNIVERSITY STATION R7000 10100 BURNET RD, EME133 AUSTIN, TX 78758 Focus for the Future – Leveraging UT Expertise and Resources Sugar Cane 2005 Algae 2007 Education and Services + 2013 - 2015 • Establish Industry Analytical Standards for measuring algal biomass and products • Discussed with the international community • Follow-up expected with NIST, DOE, and EU • Industrial Consortium • Proposed by current sponsors • Assessing feasibility • Processing R & D • Collaborative with Industrial Sponsors Algae – Providing Analytical Protocols and Measurement Standards to the Algae Industry With $ millions invested and 100’s of developers, why is a positive EROI always 5 years away? • Non-standardized measurements of virtually every parameter – each method can produce wildly different results on the same culture • Cell density • Cell count • Gravity settling • Dry weight with and without centrifugation to remove salts • Filtering Algae – Providing Analytical Protocols and Measurement Standards to the Algae Industry With $ millions invested and 100’s of developers, why is a positive EROI always 5 years away? • Total oil content measurement challenges • Nile red (non-specifically binds to a variety of neutral lipid compounds) • Gravimetric (anything recovered in a particular solvent – non-specific to fuel or high value products) • HPLC/MS – detailed quantitative lipid profiling of specific lipid classes and species of interest • GC/MS – detailed quantification of fatty acids • Because UT has developed and published advanced analytical tools that define and quantify specific algae products for the industry, UT is in a unique position to help define analytical measurement standards that the industry can adopt Algae – Providing Analytical Protocols and Measurement Standards to the Algae Industry With $ millions invested and 100’s of developers, why is a positive EROI always 5 years away? • Incomplete knowledge of membrane and cell wall electrical and mechanical properties • Many inefficient cell breaking approaches are utilized in the industry instead of an optimized approach • Oil-in-water emulsions form from broken algae debris – affects recovery of the oil/products • Because UT/CEM has 40+ years expertise in assessing electrical and mechanical properties of materials, UT is in a unique position to characterize algal membrane and cell wall dynamics during processing to optimize recovery of algal products The UT Approach to Define Analytical Protocols and Measurement Standards for the Algae Industry • Understand cell structures and physiology – Tensile strength and electromechanical properties of cell membranes and walls – Physiological defense mechanisms against predators – Impediments to oil extraction • Standards for oil content and growth – Government-industry-university metrology consortium • International participation beneficial – Standard reference “algae cell” to validate metrology Summary • Algae represent a viable source of alternative fuels, plus high-value products • CEM has developed and transferred to industry costeffective processing technologies and continues to develop additional processing capabilities • CEM is educating the algae industry • CEM is in a unique position to lead the effort to drive the industry into profitability by developing measurement standards for the industry