1. A Winter Barley Biorefinery Producing Advanced Biofuels and Coproducts K. Hicks*, N. Nghiem, A. Boateng, C. Mullen, D. Johnston, and R. Moreau Sustainable Biofuels and CoProducts Research Eastern Regional Research Center ARS, USDA 600 E. Mermaid Lane Wyndmoor, PA 19038 kevin.hicks@ars.usda.gov

2. Apply name/department/presentation title in 'header and footer' Number of Ethanol Plants, Locations, and Their Capacities as of 3/3/2011 204 Plants with 14.1 Billion Gallons Capacity 5 Plants under Construction will Provide another 0.5 Billion Gallons Total Capacity When Completed = 14.6 Bil. Gal. 14.6 Billion Gallons meets about 10% of our total transportation fuel needs! Corn is Still the #1 Feedstock Source: RFA

3. Apply name/department/presentation title in 'header and footer' The 2007 Energy Independence and Security Act Requires Aggressive Increase in Advanced Biofuels! * Advanced biofuels is renewable fuel other than ethanol derived from corn starch that is derived from renewable biomass, and achieves a 50 percent greenhouse gas (GHG) emissions reduction (compared to gasoline). *

4. Apply name/department/presentation title in 'header and footer' 4 Cellulosic Ethanol Outstanding Potential Uses Non Food Feedstocks Still Major Research Challenges to Solve Still 5 years away from commercial viability? Pyrolysis and Gasification-Based Bio-Fuels Outstanding Potential Uses Non Food Feedstocks Still Major Research Challenges to Solve Still a few years away from commercial viability How will we meet these goals?

5. Apply name/department/presentation title in 'header and footer' The 2007 Energy Independence and Security Act Requires Aggressive Increase in Advanced Biofuels! * Advanced biofuels is renewable fuel other than ethanol derived from corn starch that is derived from renewable biomass, and achieves a 50 percent greenhouse gas (GHG) emissions reduction (compared to gasoline). *

6. Apply name/department/presentation title in 'header and footer' 6 Is There a Simpler Way? Wouldn’t it be great if there were another feedstock we could use right now to make Advanced Biofuels? One that we could convert to advanced fuel ethanol and valuable food and feed products One that wouldn’t compete with food crop production One that wouldn’t harm the environment

7. Apply name/department/presentation title in 'header and footer' 7 There is Such a Crop and it is Winter Barley! These “barley belts” can provide feedstock for ethanol plants outside the corn belt where transportation fuels and economic development are needed!

8. Apply name/department/presentation title in 'header and footer' 8 Why Winter Barley For Fuel Ethanol ? Provides feedstock for ethanol plants outside the Corn Belt Farmers on the East Coast of the United States and other areas with mild winters can grow barley as a winter crop, allowing double cropping with soy or corn (More grain on same acreage) Winter barley is grown on “winter fallow ground” and doesn’t compete with food production thus there are no Indirect Land Use Change effects. Winter barley acts as a cover crop, preventing soil and nutrient losses to the environment- this is critical for sustainability of soil and water. Especially important for the Delaware and Chesapeake Bays. Higher protein and digestible amino acids than corn, especially lysine means that barley DDGS should sell at a premium relative to other grain DDGS.

9. Apply name/department/presentation title in 'header and footer' 9 Major Challenges with Barley for Ethanol Production in 2001 Abrasive nature of hull – destructive to grain handling and grinding equipment Low starch content (~50-55%) compared to corn’s (~70%) – results in low ethanol yields High viscosity of mash due to beta-glucans – makes ethanol production difficult and expensive and limits the feed use of the ethanol co-products, DDGS to primarily ruminant animals Phytoliths

10. Apply name/department/presentation title in 'header and footer' 10 ERRC/ARS Created A Barley Research Program to Solve These Technical Issues

11. Apply name/department/presentation title in 'header and footer' THOROUGHBRED DOYCE We Worked with Breeders at Virginia Tech to Develop Better Barley Varieties Prof. Carl Griffey Wynse Brooks Barley Breeder Griffey, C., Brooks, W., Kurantz, M., Thomason, W., Taylor, F., Obert, D., Moreau, R., Flores, R., Sohn, M., and Hicks, K. Grain composition of Virginia winter barley and implications for use in feed, food, and biofuels production. Journal of Cereal Science. 51: 41–49. 2010.

12. Apply name/department/presentation title in 'header and footer' Composition of Barley is Critical for Fuel Ethanol Production COMPONENT (%, DWB) “NOMINI”- HULLED FEED BARLEY (POOR) “THOROUGHBRED” V.T. ELITE HULLED (BETTER) “EVE”- V.T. ELITE HULL-LESS (BEST) STARCH54.859.963.8  -GLUCAN 5.03.94.1 PROTEIN8.87.610.0 OIL2.51.9 ASH2.3 1.7 NDF26.017.211.0 TEST WT (LB/BU)48.152.960.8

13. Apply name/department/presentation title in 'header and footer' What about those Abrasive Hulls? Many of the barley varieties developed were hull-less varieties We also developed methods to remove the abrasive hulls and to produce starch-enriched fractions for ethanol production. 13 Flores, R.A., Hicks, K.B., Eustace, D.W., and Phillips, J.G. High- starch and high-ß-glucan barley fractions milled with experimental mills. Cereal Chem. 82(6):727-733. 2005.

14. Apply name/department/presentation title in 'header and footer' How Did We Solve the  -Glucan Viscosity Issue? By Working with Genencor, a Danisco Division, to Develop Better Enzymes and Processes

15. Apply name/department/presentation title in 'header and footer' 15 The State-of-the-Art with  -Glucanase Enzymes in 2001 Commercial Enzyme Preparations Contained Mixtures of Enzymes that Partially Degraded Viscous High Molecular Weight  -Glucans, Producing Low Viscosity Oligosaccharides in the Process.

16. Apply name/department/presentation title in 'header and footer' 16 Our Studies Showed How to Take it a Step Further We found that complete conversion of  -glucan to glucose can be achieved with use of additional amounts of a key enzyme,  -Glucosidase This completely hydrolyses  -glucan producing additional fermentable glucose, Increasing Yields Barley containing 65% starch and 5%  -glucan should be equivalent to corn’s 70% starch! This is What Provides the “Edge” to the Barley EDGE Process

17. Apply name/department/presentation title in 'header and footer' 17 Fresh water Milled Barley Evaporation condensate SPEZYME® Xtra OPTIMASH™ BG 30-33°C pH 3.8-4.2 Steam 85-90°C pH 5.2 85-90°C pH 5.2 FERMENZYME® L-400 58 - 60°C 60 min Thin stillage OPTIMASH™ TBG OPTIMASH™ BG Urea Barley EDGE* Process * Enhanced Dry Grind Enzymatic Pre-liquefaction Liquefaction SSF  -gluco- sidase Nghiem, N.P et al. Biotechnology for Biofuels. 3:8. 2010.

18. Apply name/department/presentation title in 'header and footer' The EDGE Process Accomplishes Three Major Goals Reducing Viscosity of Mash Increasing Ethanol Yields Improves composition and quality of DDGS Nghiem, N.P., Hicks, K.B., Johnston, D.B., Senske, G., Kurantz, M., Li, M., Shetty, J., and Konieczny-Janda, G. Production of ethanol from winter barley by the EDGE (enhanced dry grind enzymatic) process. Biotechnology for Biofuels 3:8. 2010. Nhuan P. Nghiem, Edna C. Ramírez, Andrew J. McAloon, Winnie Yee, David B. Johnston, Kevin B. Hicks. Economic analysis of fuel ethanol production from winter hulled barley by the EDGE (Enhanced Dry Grind Enzymatic) process. Bioresource Technology 102 (2011) 6696–6701. Nhuan P. Nghiem Frank Taylor Kevin B. Hicks David B. Johnston Jay K. Shetty. Scale-up of ethanol production from winter barley by the EDGE (enhanced dry grind enzymatic) process in fermentors up to 300 liters. Applied Biochemistry and Biotechnology, In Press 2011.

19. Apply name/department/presentation title in 'header and footer' 19 41 Growing winter barley for ethanol feedstock does not compete with food production and it actually improves soil and water quality while giving farmers a new source of income! Premium DDGS The EDGE Process The Winter Barley Ethanol Process Contact: kevin.hicks@ars.usda.gov

20. Apply name/department/presentation title in 'header and footer' 20 In 2008, Osage Bio Energy Began Building A Winter Barley Ethanol Plant in Hopewell VA

21. First US Winter Barley Ethanol Facility Products To Be Produced: 65 million gallons of fuel-grade ethanol 250 thousand tons of high-value DDGS 65 thousand tons of barley hull fuel pellets 170 thousand tons of carbon dioxide

22. Apply name/department/presentation title in 'header and footer' Can the Winter Barley Ethanol Process make Money?? 22 1 X 48 lb bushel of barley = $2.00 2 gal ethanol at $2.00 per gallon = $4.00 DDGS 16 lb DDGS = $0.64 $2.64 Value Added 1 X 48 lb bushel of barley = $6.50 2 gal ethanol at $2.50 per gallon = $5.00 DDGS 16 lb DDGS = $1.90 $0.40 Value Added

23. Apply name/department/presentation title in 'header and footer' YES! Can We Expand this Fuel Ethanol Process into a Biorefinery to Produce Additional Biofuels and CoProducts and Additional Revenue? 23

24. Apply name/department/presentation title in 'header and footer' 24 CoProducts from Hulls Cellulosic Ethanol

25. Apply name/department/presentation title in 'header and footer' 25 Ammonia recycling Barley (100 lb) 60% starch (0% moisture) Recombinant microorganism (example: KO11) Roller Mill Aspirator Starch 55 lb* 15 lb Hulls SaccharificationFermentation Glucose (67 lb)  -Glucan (4 lb) Protein (10 lb) Others (14 lb) 32.4 lb of EtOH Saccharification SAA Reactor 15 lb Co-Fermentation Making Ethanol From Barley Starch and Barley Hulls SSCF 16%* Approx. 11% ethanol yield increase by utilization of hulls. 16%* increase if 100% conversion Starch Ethanol 35.9 lb (5.5 gallons) of Washing (ex.counter current leaching) Enzyme + Starch (5 lb)* Destarching Starch 60 lb 85 lb Kernels Cellulosic Ethanol 3.5* lb of EtOH Kim, T.H., Taylor, F., and Hicks, K.B. Bioethanol production from barley hull using SAA (soaking in aqueous ammonia) pretreatment. BioResource Technology 99:5694 5702. 2008.

26. Apply name/department/presentation title in 'header and footer' 26 Thermochemically Derived BioFuels and CoProducts Conversion of Barley Hulls, Straw, and DDGS by Pyrolysis can yield valuable Bio-Oil and Bio-Char Bio-Oil and Bio-Char Hulls, Straw, DDGS Green Gasoline and Diesel Improves soil fertility and sequesters carbon Boateng, A.A., K.B. Hicks, R.A. Flores and A. Gutsol. Pyrolysis of hull-enriched byproducts from scarification of barley (Hordeum vulgare L.). J. Analy. & Appl. Pyrolysis, 78: 95-103 (2007). Mullen, C.A., A.A. Boateng, N.M. Goldberg, K.B. Hicks, and R. Moreau. Analysis and Comparison of bio-oil produced by fast pyrolysis from three barley biomass/byproduct streams. Energy & Fuels 24:699–706 (2010).

27. Apply name/department/presentation title in 'header and footer' YES! Can We Also Make High Valued Nutraceutical CoProducts? 27

28. Apply name/department/presentation title in 'header and footer' 28 Anatomy of a Barley Kernel T = Thoroughbred Hulled Barley Kernel g = Germ (Embryo) e = Endosperm h = Hull Images: Paul Pierlott, ERRC Starch, Protein,  -glucan, Tocotrienols Triacylglycerols, Tocopherols, Phytosterols Cellulose, Hemicellulose, Lignin, Ash

29. Apply name/department/presentation title in 'header and footer' 29 Nutraceuticals in Barley Phytosterols Anticholesterolemic Tocopherols Antioxidants Vitamin E Tocotrienols Antioxidants Anticancer Anticholesterolemic

30. Apply name/department/presentation title in 'header and footer' 30 Bio-Refinery CoProducts: Fractionation of Kernels for Value Added Fractions – Scarification and Nutraceutical Lipid Extraction Scarification Whole Barley Kernels ~2% oil Abraded Kernels < 2% oil Fines and Germ Fragments 3-9% oil Barley oil can be extracted from whole kernels or, more efficiently, from lipid enriched abraded fines and germ fragments. Barley oil is rich in Phytosterols (1.2 – 10g/100 gram oil), Tocopherols (T) (0.15-0.28 g/100 gram oil), and Tocotrienols (T3) (0.1-0.3 g/100 gram oil). Barley Oil has the highest levels of Tocotrienols reported for a natural oil: ( Rice Bran and Palm oils only have 0.05-0.08 g T3 /100 gram oil) Moreau, R.A., Flores, R., and K.B. Hicks, Cereal Chem. 84:1-5, 2007. Moreau, R.A., Wayns, K.E., Flores, R.A., and Hicks, K.B. Cereal Chem. 84:587-592. 2007.

31. Apply name/department/presentation title in 'header and footer' 31 41 Growing winter barley for ethanol feedstock does not compete with food production and it actually improves soil and water quality while giving farmers a new source of income! High starch fractions are used for fermentation, and Fuel Ethanol Production plus Premium DDGS Low starch fractions (high-protein, high-Tocol, high-  -glucan) for health-promoting, obesity-fighting, foods and nutraceuticals FRACTIONATION PROCESSES High fiber (hulls) and straw from barley can be used to make cellulosic ethanol and pyrolysis oil for producing “green” transportation fuels. The Winter Barley BioRefinery Vision Contact: kevin.hicks@ars.usda.gov ?

32. Apply name/department/presentation title in 'header and footer' 32 ACKNOWLEDGEMENTS ARS John Nghiem, Andy McAloon, Winnie Yee, Edna Ramirez, Frank Taylor, David Johnston, Rolando Flores, Bob Moreau, Gerry Senske, Akwasi Boateng, Charles Mullen, Mike Kurantz, Robyn Moten, Mike Powell, Jhanel Wilson. Virginia Tech Dan Brann, Carl Griffey, Wynse Brooks, Wade Thomason, Bruce Beahm, and Mark Vaughn Genencor, a Danisco Division Bruce Strohm, Jay Shetty, Mian Li, Gerhard Konieczny-Janda, Brad Paulson, Pauline Tenuissen, and Bob Randle Osage Bio Energy Craig Shealy, Joel Stone, Pat Simms, Eric Lee, Hank Bisner, Bill Scruggs, Tim Richter, Earl Spruill, John Warren. Katzen International -- Phil Madson