1. Consortium for Biomass Refining Based on Leading Pretreatment Technologies Charles E. Wyman, Dartmouth College/University of California Bruce E. Dale, Michigan State University Richard T. Elander, National Renewable Energy Laboratory Mark T. Holtzapple, Texas A&M University Michael R. Ladisch, Purdue University Y. Y. Lee, Auburn University Mohammed Moniruzzaman, Genencor International John N. Saddler, University of British Columbia International Symposium on Alcohol Fuels San Diego, California September 26, 2005 Biomass Refining CAFI

2. Topics Covered Need for new transportation fuels Overview of cellulosic biomass Conversion of cellulosic biomass to ethanol Importance of pretreatment Biomass Refining Consortium on Applied Fundamentals and Innovation - CAFI Structure of recently completed USDA IFAFS Project Structure of in progress DOE OBP Project Biomass Refining CAFI

3. Where is a New Energy Source Needed in United States? U.S. energy production and demand are nearly balanced for all but one energy source: petroleum We use far more petroleum than we produce – >60% imported Petroleum is the largest energy source in U.S. supplying ~38.5% Biomass Refining CAFI

4. Why Are Transportation Fuels Important Targets? About 2/3 of petroleum goes to transportation Transportation is almost totally dependent on petroleum (~96.4% in 2000) The largest amount of carbon dioxide comes from transportation, ~32.8% Biomass Refining CAFI

5. Ethanol Production in Brazil and the United States United States Brazil from cane sugar from starch crops (e.g., corn) Biomass Refining CAFI

6. Focus: Cellulosic Biomass - Abundant, Inexpensive Existing resources –Agricultural wastes Sugar cane bagasse Corn stover and fiber –Forestry wastes Sawdust –Municipal wastes Waste paper Yard waste –Industrial waste Pulp/paper sludge Future resources –Dedicated crops Herbaceous Woody Not sugar or starch crops such as used for making ethanol in Brazil and the U.S. respectively Biomass Refining CAFI

7. Louisiana Rice Hulls Pile Biomass Refining CAFI

8. Florida Sugarcane Bagasse Biomass Refining CAFI

9. Energy Crops Switchgrass harvested annually or biannually Willow coppice harvested at age 3 or 4 Hybrid Poplar harvested at age 5 to 10 Courtesy of L. Wright, ORNL Biomass Refining CAFI

10. Billion Ton Supply of Cellulosic Biomass DOE and USDA recently estimated 1.3 billion tons of cellulosic biomass could be available Includes 368 million dry tons from forests and 998 million dry tons from agriculture Biomass Refining CAFI

11. Cost of Cellulosic Biomass vs Petroleum Biomass Refining CAFI

12. Challenge: How Do You Put Low Cost Biomass in Your Car? Biomass Refining CAFI

13. Cellulosic Biomass Composition Cellulose 45% Hemicellulose 30% Lignin 15% Other 10% Herbaceous Energy Crops Cellulose 43% Hemicellulose 27% Lignin 17% Other 13% Agricultural Residues Municipal Solid Waste Ash 15% Lignin 10% Hemicellulose 9% Other carbohydrates 9% Protein 3% Other 9% Cellulose 45% Cellulose 45% Hemicellulose 25% Lignin 22% Extractives 5% Ash 3% Woody Crops Biomass Refining CAFI

14. Breakdown hemicellulose to sugars Make enzymes, breakdown cellulose to glucose, and ferment all sugars Enzymatic Conversion of Cellulosic Biomass to Ethanol Biological steps: Cellulase production Hydrolysis Fermentation Feedstock Production Pretreatment Ethanol recovery Residue processing Utilities Fuel ethanol Process effluents Exported electricity Process boundaries Lignin, etc Process Heat, Electricity Biomass Refining CAFI

15. Benefits of Cellulosic Ethanol Environmental –Little if any greenhouse gas emissions –Solid waste disposal –Can improve air quality –Low impact biomass crops Economic –Abundant, inexpensive, domestic feedstock –Low cost potential without subsidies –Agricultural and manufacturing employment Energy –Secure resource available for most countries Biomass Refining CAFI

16. Significant Progress in Enzyme Based Technology for Cellulosic Ethanol Corn EtOH Price Based on historic estimates by NREL 1980Now

17. Key to Advances To Date in Cellulosic Ethanol Technology Overcoming the recalcitrance of cellulosics –Improved pretreatment to increase yields from hemicellulose and cellulose –Improved cellulase enzymes to increase rates from cellulose, reduce enzyme use –Integrated systems to improve rates, yields, concentrations of ethanol (SSF) Overcoming the diversity of sugars –Recombinant organisms ferment all five sugars to ethanol at high yields Biomass Refining CAFI

18. Biological Conversion of Cellulosics Pretreatment Product recovery Cellulose Hydrolysis and Sugar Fermentation Cellulosic biomass Prepared biomass Fermentation broth Ethanol Cellulase Production Cellulase enzyme Organism Chemicals Lignin, ash, etc for boiler, chemicals Biomass Refining CAFI

19. Biological Conversion of Cellulosics Pretreatment Product recovery Cellulose Hydrolysis and Sugar Fermentation Cellulosic biomass Prepared biomass Fermentation broth Ethanol Cellulase Production Cellulase enzyme Organism Chemicals Lignin, ash, etc for boiler, chemicals Biomass Refining CAFI

20. Cellulosic Biomass Pretreatment Needs High cellulose accessibility to enzymes High sugar yields from hemicellulose Low capital cost – low pressure, inexpensive materials of construction Low energy cost Low degradation Low cost and/or recoverable chemicals Biomass Refining CAFI

21. Cellulosic Biomass Pretreatment Needs High cellulose accessibility to enzymes High sugar yields from hemicellulose Low capital cost – low pressure, inexpensive materials of construction Low energy cost Low degradation Low cost and/or recoverable chemicals A large number of pretreatment technologies have been studied, but only a few show promise Biomass Refining CAFI

22. Biomass Refining Consortium for Applied Fundamentals and Innovation (CAFI) Organized in late 1999 Included top researchers in biomass hydrolysis from Auburn, Dartmouth, Michigan State, Purdue, NREL, Texas A&M, UBC, U. Sherbrooke l Mission: l Develop information and a fundamental understanding of biomass hydrolysis that will facilitate commercialization, l Accelerate the development of next generation technologies that dramatically reduce the cost of sugars from cellulosic biomass l Train future engineers, scientists, and managers. Biomass Refining CAFI

23. CAFI USDA IFAFS Project Overview Multi-institutional effort funded by USDA Initiative for Future Agriculture and Food Systems Program for $1.2 million to develop comparative information on cellulosic biomass pretreatment by leading pretreatment options with common source of cellulosic biomass (corn stover) and identical analytical methods –Aqueous ammonia recycle pretreatment - YY Lee, Auburn University –Water only and dilute acid hydrolysis by co-current and flowthrough systems - Charles Wyman, Dartmouth College –Ammonia fiber explosion (AFEX) - Bruce Dale, Michigan State University –Controlled pH pretreatment - Mike Ladisch, Purdue University –Lime pretreatment - Mark Holtzapple, Texas A&M University –Logistical support and economic analysis - Rick Elander/Tim Eggeman, NREL through DOE Biomass Program funding Completed in 2004 Biomass Refining CAFI

24. Feedstock: Corn Stover NREL supplied corn stover to all project participants (source: BioMass AgriProducts, Harlan IA) Stover washed and dried in small commercial operation, knife milled to pass ¼ inch round screen Glucan36.1 % Xylan21.4 % Arabinan3.5 % Mannan1.8 % Galactan2.5 % Lignin17.2 % Protein4.0 % Acetyl3.2 % Ash7.1 % Uronic Acid3.6 % Non-structural Sugars1.2 % Biomass Refining CAFI

25. USDA IFAFS Project Tasks Apply leading pretreatment technologies to prepare biomass for conversion to products Characterize resulting fluid and solid streams Close material and energy balances for each pretreatment process Determine cellulose digestibility and liquid fraction fermentability Compare performance of pretreatment technologies on corn stover Biomass Refining CAFI

26. Presentations and Publications of CAFI Results Team presentations at –2004 Annual Meeting of the American Institute of Chemical Engineers, Austin, Texas, November 11 –2003 Annual Meeting of the American Institute of Chemical Engineers, San Francisco, California, November 20 –25th Symposium on Biotechnology for Fuels and Chemicals, Breckenridge, Colorado, May 7, 2003 –2002 Annual Meeting of the American Institute of Chemical Engineers, Indianapolis, Indiana, November 4 –24th Symposium on Biotechnology for Fuels and Chemicals, Gatlinburg, Tennessee, April 28, 2002 Mosier N, Wyman CE, Dale B, Elander R, Lee YY, Holtzapple M, Ladisc1 M. 2005. “Features of Promising Technologies for Pretreatment of Lignocellulosic Biomass,” BioResource Technology 96(6): 673-686 Special December issue of Bioresource Technology reports USDA IFAFS findings in several papers including joint papers to introduce project and summarize results Biomass Refining CAFI

27. DOE Office of the Biomass Program Project: April 2004 Start Funded by DOE Office of the Biomass Program for $1.88 million through a joint competitive solicitation with USDA Using identical analytical methods and feedstock sources to develop comparative data for corn stover and poplar Determining more depth information on –Enzymatic hydrolysis of cellulose and hemicellulose in solids –Conditioning and fermentation of pretreatment hydrolyzate liquids –Predictive models Added University of British Columbia to team through funding from Natural Resources Canada to –Capitalize on their expertise with xylanases for better hemicellulose utilization –Evaluate sulfur dioxide pretreatment along with those previously examined: dilute acid, controlled pH, AFEX, ARP, lime Augmented by Genencor to supply enzymes Biomass Refining CAFI

28. DOE OBP Project Tasks Biomass Refining CAFI Corn stover and poplar pretreated by leading technologies to improve cellulose accessibility to enzymes Conditioning methods developed as needed to maximize fermentation yields by a recombinant yeast, the cause of inhibition determined, and fermentations modeled Cellulose and hemicellulose in pretreated biomass enzymatically hydrolyzed, as appropriate, and models developed to understand the relationship between pretreated biomass features, advanced enzyme characteristics, and enzymatic digestion results Capital and operating costs estimated for each integrated pretreatment, hydrolysis, and fermentation system and used to direct research

29. CAFI Project Advisory Board Serve as extension agents for technology transfer Provide feedback on approach and results Meet with team every 6 months Quang Nguyen, Abengoa Bioenergy Mat Peabody, Applied CarboChemicals Gary Welch, Aventinerei Greg Luli, BC International Paris Tsobanakis, Cargill Robert Wooley, Cargill Dow James Hettenhaus, CEA Kevin Gray, Diversa Paul Roessler, Dow Susan M. Hennessey, DuPont Michael Knauf, Genencor Don Johnson, GPC (Retired) Dale Monceaux, Katzen Engineers Kendall Pye, Lignol Mark Stowers, MBI Richard Glass, National Corn Growers Association Bill Cruickshank, Natural Resources Canada Joel Cherry, Novozymes Ron Reinsfelder, Shell Carl Miller, Syngenta Carmela Bailey, USDA Don Riemenschneider, USDA Biomass Refining CAFI

30. ISAF XV Overviews of CAFI Research Pretreatment – Bruce Dale, Michigan State Enzymatic hydrolysis – Charles Wyman, Dartmouth/University of California Logistics and economics – Richard Elander, NREL Focus on results from recently completed USDA-funded project with some new results from the DOE-funded project Biomass Refining CAFI

31. Acknowledgments US Department of Agriculture Initiative for Future Agricultural and Food Systems Program, Contract 00-52104-9663 US Department of Energy Office of the Biomass Program, Contract DE-FG36- 04GO14017 Natural Resources Canada Biomass Refining CAFI

32. Insanity is doing what you always have always been doing and expecting different results Biomass Refining CAFI

33. Questions? Biomass Refining CAFI