1. Creating a Secure and Sustainable Future Defining an Achievable Means of Meeting our Food, Fuel and Feed Needs Domestically Martha Schlicher, Ph.D. National Corn-to-Ethanol Research Center

2. Definitions Ethanol A two carbon alcohol produced most easily produced by conventional yeast when they consume single six carbon sugars. Used as spirit or fuel. Sucrose Two six carbon sugars (glucose and fructose) linked together. Table sugar. Found in sugar cane Glucose The sugar in your blood Exists as chains of glucose (e.g. starch or cellulose) or glucose with other sugars Starch Chains of the six carbon sugar glucose present in corn or potatoes that can be easily broken down to free glucose and fermented to ethanol. A carbohydrate. Cellulose Chains of the six carbon sugar glucose present in fiber/biomass. Potential to convert to ethanol. A carbohydrate. Hemicellulose Chains of five and six carbon sugars present in fiber/biomass. Potential to convert to ethanol. A carbohydrate.

3. Robert Shleser, 1994 Starch Cellulose Lignocellulose Sucrose Lignin Examples

4. TIME M Barrels of Oil Displaced/Yr THE POTENTIAL ETHANOL PIPELINE NEAR TERM 10% MID TERM 20% LONG TERM 40% B Gallons of Ethanol Produced/Yr GLUCOSE FROM STARCH & CORN KERNEL CELLULOSE SUCROSE FROM SUGARCANE/BEETS GLUCOSE FROM STARCH CORN KERNEL & AVAILABLE BIOMASS CELLULOSE (wood chips, waste) PENTOSES FROM AVAILABLE BIOMASS HEMICELLULOSE GLUCOSE FROM STARCH CORN KERNEL & NEW CELLULOSE BIOMASS PENTOSES FROM NEW BIOMASS HEMICELLULOSE 2005-20152016-2030 2031-2040 4 115 21 MBD Oil in 05; Gasoline only; Use held at 05 20 772 40 1554

5. ABUNDANT & AVAILABLE ECONOMICAL Corn Starch Switch Grass US Biomass Sources Paper Sugar Cane Cottonwoods Stover Wood Chips Corn Fiber NEAR TERM Wheat Barley

6. NEAR TERM STARCH 1980's --175 plants (20K to 7M gallons) 1990's -- 33 plants (1.5M to 30 M gallons) 2000's -- 91 plants (30M to 110M gallons) Important to Remember Secure the Base while Building the Future

7. TIME Barrels of Oil Displaced RESOURCING THE ETHANOL PIPELINE NEAR TERM 10% MID TERM 20% LONG TERM 40% Gallons of Ethanol Produced AGRONOMIC PRODUCTIVITY DDGS = CORN PROCESS PRODUCTIVITY ENERGY USE NEW COPRODUCTS GLUCOSE FROM STARCH CORN KERNEL & AVAILABLE BIOMASS CELLULOSE (wood chips, waste) PENTOSES FROM AVAILABLE BIOMASS HEMICELLULOSE GLUCOSE FROM STARCH CORN KERNEL & NEW CELLULOSE BIOMASS PENTOSES FROM NEW BIOMASS HEMICELLULOSE 2005-20152016-2030 2031-2040 115 772 1554 4 20 40

8. NEAR TERM STARCH

9. e S n NEAR TERM STARCH

10. 2004 US Corn Utilization FEED: 56.4% EXPORT: 18.5% FUEL: 11.7% FOOD: 6.7% HFCS: 5.2% 80.9 Million Acres Harvested - 11.8 Billion Bushels NEAR TERM STARCH

11. Ethanol Production Locations

13. Critical Near Term Research Increased Ethanol Yield Per Acre Reduced Energy Use per Gallon Agronomic Productivity High Extractable/Fermentable Hybrids Custom Hybrids Process Modifications Cold Cook/Enzyme advancements Degerm/Debran Corn Kernel Fiber to Ethanol Coproduct Optimization and Segregation DDGS Composition, Quality, Transport Wet Distillers/Plant Location Oil Separation for Biodiesel Fermentation/Starch to Sugar Ethanol Isolation Organism Optimization Ethanol Separation Cogeneration Mining Process Energy and Streams

14. Critical Near Term Research Increased Ethanol Yield Per Acre Reduced Energy Use per Gallon Agronomic Productivity High Extractable/Fermentable Hybrids Custom Hybrids Process Modifications Cold Cook/Enzyme advancements Degerm/Debran Corn Kernel Fiber to Ethanol Coproduct Optimization and Segregation DDGS Composition, Quality, Transport Wet Distillers/Plant Location Oil Separation for Biodiesel Fermentation/Starch to Sugar Ethanol Isolation Organism Optimization Ethanol Separation Cogeneration Mining Process Energy and Streams

15. 2004 US Corn Utilization FEED: 56.4% EXPORT: 18.5% FUEL: 11.7% FOOD: 6.7% HFCS: 5.2% 80.9 Million Acres Harvested - 11.8 Billion Bushels NEAR TERM STARCH

16. One Bushel (56 lbs) of Corn Entering Dry Grind Ethanol Plant STARCH PROTEIN OIL FIBER Pounds Per Bushel of Corn – Dry Weight Assume corn 15% moisture content 37.2 lbs 4.98 lbs 2.24 lbs 4.98 lbs 0 5.01 lbs 1.74 lbs ETHANOL CO 2 DDGS – 16 -18 lbs Products from one Bushel of Corn Leaving Dry Grind Ethanol Plant 0 0 2.75 gallons 18.0 lbs 3.74 lbs NEAR TERM STARCH

17. 1.38 B Bushels of Corn Entering Ethanol Plants STARCH PROTEIN 37.2 lbs 6.88 B lbs 0 7.04 B lbs ETHANOL CO 2 Results in ………… 0 0 2.75 gallons 18.0 lbs Containing Animal Feed NEAR TERM STARCH

18. Amounts of feedstocks to produce 10 ml ethanol 500 ml graduated cylinders used for comparisons CornFiberDDGStover ethanol Moisture Content Corn15% Fiber46% DDG64% Stover 5% NEAR TERM CORN KERNEL FIBER

19. Corn Kernel Cellulosics Near Term Technology Validation No incremental supply chain Costs Potential 10% Yield increase 4.5 M gal Ethanol per plant Annually Minimal incremental capital DDGS weight reduced 44% No increase in corn acres NEAR TERM

20. Robert Shleser, 1994 Starch Cellulose Lignocellulose Sucrose Lignin Examples

21. Representative Corn Component Composition ComponentStoverCobsFiber Glucan (%) Xylan (%) Arabinan (%) Lignin (%) Protein (%) Fat (%) 40.9 21.5 1.8 11.0 7.2 1.3 39.4 28.4 3.6 7.0 3.2 0.7 37.2 17.6 11.2 7.8 11.0 2.5 Wyman; Corn Chemistry and Technology, 2 nd edtn.

22. Selected Pretreatment Strategies Acid Base PretreatmentPentosesInhibitors Strong Acid +++ Dilute Acid +++ Hot Water -+ AFEX -- Alkaline Peroxide --

23. Yeast metabolism & nutrition Sugar and carbohydrates metabolised by Saccharomyces Species Sugar sources Glucose Fructose Mannose Galactose Sucrose Maltose Maltotriose (partially) Raffinose Dextrine (partially ) Sugar and carbohydrates not metabolysed by saccharomyces species but metabolised by species of other yeast genera Xylose (e.g. Candida shehatae and Pichia stipitis) Cellobiose (e.g. Scwanniomyces castelli and Candida) Lactose ((e.g. Candida curvata and Kluyveromyces fragilis) Starch (e.g. Schwanniomyces castellii and Saccharomyces fibuligera) Inulin (e.g. Kluyveromyces marxianus) Metabolites Glucose Fructose Mannose Galactose Glucose + Fructose 2 Glucoses 3 Glucoses Raffinose Dextrine (partially) Ethanol

24. Recombinant Microorganisms for Fermentation of Mixed Sugars to Ethanol Recombinant organisms are now available Recombinant Escherichia coli Recombinant Saccharomyces Recombinant Zymomonas Recombinant Klebsiella oxytoca Timid Commercialization prospects BCI with recombinant E. coli Iogen with recombinant Saccharomyces

25. Utilization of Biomass for Production of Fuel Ethanol Corn Fiber Ethanol Recovery Pretreatment Enzymatic Saccharification Fermentation

26. Representative Corn Component Composition ComponentStoverCobsFiber Glucan (%) Xylan (%) Arabinan (%) Lignin (%) Protein (%) Fat (%) 40.9 21.5 1.8 11.0 7.2 1.3 39.4 28.4 3.6 7.0 3.2 0.7 37.2 17.6 11.2 7.8 11.0 2.5 Wyman; Corn Chemistry and Technology, 2 nd edtn.

27. 1993 Corn Kernel Fiber Solved

28. Strategy 1 Design a method to CONVERT ALL possible sugars in kernel, maximum increase In ethanol would be 0.328 gallons/bushel. Weight, lbs/bushel*Ethanol, gal/bushel* Starch33.932.93 Hemicellulose2.590.228 Cellulose1.160.100 *From Gulati, et al., 1996 Bioresource Technology Math for 50 mgy plant:0.328 gal/2.7 gal is 12% increase 0.10 gal/2.7 gal is 3.7% increase

29. ABUNDANT & AVAILABLE ECONOMICAL Corn Starch Switch Grass US Biomass Sources Paper Sugar Cane Cottonwoods Stover Wood Chips Corn Fiber MID TERM Wheat Barley

30. TIME Barrels of Oil Displaced RESOURCING THE ETHANOL PIPELINE NEAR TERM 10% MID TERM 20% LONG TERM 40% Gallons of Ethanol Produced AGRONOMIC PRODUCTIVITY DDGS = CORN PROCESS PRODUCTIVITY ENERGY USE NEW COPRODUCTS GLUCOSE FROM STARCH CORN KERNEL & NEW CELLULOSE BIOMASS PENTOSES FROM NEW BIOMASS HEMICELLULOSE 2005-20152016-2030 2031-2040 AVAILABLE BIOMASS SEPARATION AND PRETREATMENT OPTIMIZED FIVE CARBON SUGAR CONVERSION OPTIMIZED MIXED SUGAR OPTIMIZED REGULATORY FRAMEWORK ESTABLISHED PROCESS EFFICIENCY OPTIMIZED 115 772 1554 4 20 40

31. Representative Corn Component Composition ComponentStoverCobsFiber Glucan (%) Xylan (%) Arabinan (%) Lignin (%) Protein (%) Fat (%) 40.9 21.5 1.8 11.0 7.2 1.3 39.4 28.4 3.6 7.0 3.2 0.7 37.2 17.6 11.2 7.8 11.0 2.5 Wyman; Corn Chemistry and Technology, 2 nd edtn.

32. Ethanol Fermentation Of Corn Fiber Hydrolysate by E. coli FBR5

33. Future Strains: Critical Traits Pentose utilization High ethanol yield and productivity Genetic and phenotypic stability Hardiness (tolerance to ethanol and inhibitors) Efficient use of multiple sugars Growth at low pH/high temperature Ease of use with current production technology

34. ABUNDANT & AVAILABLE ECONOMICAL Corn Starch Switch Grass US Biomass Sources Paper Sugar Cane Cottonwoods Stover Wood Chips Corn Fiber LONG TERM Wheat Barley

35. TIME Barrels of Oil Displaced RESOURCING THE ETHANOL PIPELINE NEAR TERM 10% MID TERM 20% LONG TERM 40% Gallons of Ethanol Produced AGRONOMIC PRODUCTIVITY DDGS = CORN PROCESS PRODUCTIVITY ENERGY USE NEW COPRODUCTS IDEAL BIOMASS BY GEOGRAPHY PRODUCTION, HARVEST, And TRANSPORT BIOMASS SEPARATION PRETREATMENT CONVERSION ORGANISM OPTIMIZATION COPRODUCTS 2005-20152016-2030 2031-2040 AVAILABLE BIOMASS SEPARATION AND PRETREATMENT OPTIMIZED FIVE CARBON SUGAR CONVERSION OPTIMIZED MIXED SUGAR OPTIMIZED REGULATORY FRAMEWORK ESTABLISHED PROCESS EFFICIENCY OPTIMIZED 115 772 1554 4 20 40

37. Cost Comparison for Corn & Stover Corn Starch Corn Stover Annual Ethanol (MMGal)50 Ethanol Prod. Cost ($/gal)0.961.45 Total Prod. Costs (MM$/yr)47.872.0 Co-product Credit ($/gal)0.260.13 Feedstock costs 1 ($/gal)0.7930.51 Capital Invest. (MM$)48.0193.7 USDA, ERRC, March 2005 Hope for future 1 Corn at $2.25/bu & stover at $40/ dry ton

38. INPUTS Feedstock Sources Agricultural Production Agricultural Waste Biomass Industrial Waste Micro-Biorefineries Unique Resources vary by Local Geography OUTPUTS Process Streams Feed and Fuel Food COMPONENTS Process Fuel Labor Transportation

39. Key Areas of Focus Across a Portfolio of Time Secure the Base Grow the Future Feedstock Optimization Process Optimization Animal Feed Optimization SupplyDemand FFV Availability E85 Availability Consumer Knowledge and Demand Workforce Get the sugars out without destroying them or Creating a hostile environment Convert all sugars – effectively and efficiently Address regulated organisms Address inefficiencies of low concentration of alcohol in beer for isolation

40. Key Areas of Focus Across a Portfolio of Time Ensure there is research funding – nationally as well as locally Ensure it is spent well Balance near, mid and long term research needs Identify local resources and ensure funds are aligned to solve the technical problems you need addressed Require measurable commercial outcomes Don’t create champions Don’t starve the troops Be realistic Don’t be redundant Do reward results