Developments of Corn for Dry Grind Corn Processing

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Developments of Corn for Dry Grind Corn Processing Vijay Singh, Kent Rausch, Carl Miller and Jim Graeber University of Illinois at Urbana-Champaign Syngenta Inc. Bioenergy-I I Conference March 5-10, 5 2006 Tomar,, Portugal

Presentation Outline Conventional Dry Grind Process Ethanol Production Capacity and Growth in Dry Grind Corn Processing Industry in US Development of Corn in Dry Grind Corn Processing Effect of hybrid variability on dry grind corn process High fermentable corn hybrids Correlation between extractable starch and fermentable starch Granular starch hydrolyzing enzymes Corn hybrids with endogenous liquefaction enzymes Corn hybrids for modified dry grind corn processes Conclusions

Dry Grind Process One bushel of Corn (25.4 kg) Corn Dry Grind Facility 2.5-2.7 gal of Ethanol 15-17 lbs of DDGS Ruminant Feed

Dry Grind Process Corn Water Grinding (Hammermill) Mash Blending CO 2 Overhead product (Recycled back) Enzymes Liquefaction Yeast & Enzymes Saccharification & Fermentation Ethanol Dehydration column DDGS Stripping/Rectifying column

7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 1980 1981 1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 Ethanol Production in the US Year Billion Gallons

Ethanol Production in the US Currently 4.3 billion gal of ethanol is produced in the US every year Estimates indicate that ethanol production in the US will increase to 6.0 billion gals/yr by 2006. Most of the increase in the ethanol capacity will come from new dry grind ethanol plants Low capital cost for dry grind corn plants Tax incentives from federal and state governments Farmer co-ops ops

Developments of Corn for Dry Grind Process Hybrid Variability High fermentable corn hybrids Correlation between extractable starch and fermentable starch Corn hybrids with endogenous liquefaction enzymes Corn hybrids for modified dry grind corn processes

Hybrid Variability Hybrid variability in a dry grind corn facility is generally defined by two factors: 1. Differences in fermentability 2. Variation in the composition of DDGS

Effect of Hybrid Variability on Dry Grind Corn Process Final ethanol concentration in beer Coproduct quality Capital and Operating Cost Process fluctuations Maintenance

Extent of Hybrid Variability for Ethanol Production Extent of Hybrid Variability for Ethanol Production 16 14 12 3.26% 10 8 6 4 2 0 1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69 73 77 81 85 89 Ethanol Conc. % (v/v) Hybrids

Hybrid Specific Processing Limited number of elite line hybrids good producer yields but with good ethanol yield, too

Identifying of Hybrids with High Fermentability

Identifying of Hybrids with High Fermentability Source: http://www.monsanto.com/monsanto/us_ag/content/enhanced_value/pro_per/pro_per_corn/brochure.pdf

What Causes Hybrid Variability Variability due to genetics Starch? Protein? Other constituents? Variability due to environment (phenotype) Effect of location Effect of crop year

Correlation between Starch and Ethanol

Starch Yield and Ethanol (Dien et al 2002) Ethanol (g/l) 96 94 92 90 88 86 84 R 2 = 0.42 82 68 69 70 71 72 73 Extractable Starch Yield (% db) Dien, B.S., Bothast, R.J., Iten,, L.B., Barrios, L. and Eckhoff, S.R. 2002. Fate of Bt protein and influence of corn hybrid on ethanol production. Cereal eal Chem. 79:582-585 585

Starch Yield and Sugars (Pruiett 2002) 52 51 Sugars (%db) 50 49 48 47 R 2 = 0.048 46 45 62 63 64 65 66 67 68 69 70 Extractable Starch Yield (%db) Pruiett,, L. 2002. Unpublished Data. University of Illinois.

Starch Content and Ethanol Yield (Haefele et al 2004) Starch (w/w%, 100%DM) 77.00 76.00 75.00 74.00 73.00 72.00 71.00 70.00 69.00 68.00 67.00 R 2 = 0.6223 R 2 = 0.5198 0.3400 0.3600 0.3800 0.4000 0.4200 Ethanol Yield (CO2 g/gdm ) Haefele,, D., Owens, F., O Bryan, K. and Sevenich,, D. 2004. Selection and optimization of corn hybrids for fuel ethanol production. 21 pp. Proc. Am. Seed Trade Assoc. 59th Annual Corn and Sorghum Research Conference, Chicago, IL.

Starch Yield and Ethanol Conc. (Singh and Graeber 2005) 15 14 Ethanol (% v/v) 13 12 11 10 9 R 2 = 0.0038 8 58 60 62 64 66 68 70 Extractable Starch Yield (%db) Singh, V. and Graeber,, J.V. 2005. Effect of corn hybrid variability and planting location on ethanol yields. Trans. ASAE 48:709-714 714

Starch Yield and Ethanol Conc. (Zhan et al, 2005) 8.6 8.5 Ethanol Conc. % (v/v) 8.4 8.3 8.2 8.1 Ethanol Linear (Ethanol) R 2 = 0.25 8.0 65 66 67 68 69 70 71 72 Extratable Starch Yield (%) Zhan, X., Wang, D, Tuinstra,, M.R., Bean, S., Sieb,, P.A. and Sun, X.S. 2003. Ethanol and lactic acid production as affected by sorghum genotype and location. Industrial Crops and Products 18:245-255 255

Variability Due to Environment

Effect of Planting Location Illinois 1 Illinois 2 Iowa Michigan 16.00 Ethanol Conc. (%v/v) 14.00 12.00 10.00 8.00 6.00 4.00 2.00 0.00 Hyb 1 Hyb 2 Hyb 3 Hyb 4 Hyb 5 Hyb 6 Hybrids Singh, V. and Graeber, J.V. 2005. Effect of corn hybrid variability and planting location on ethanol yields. Trans. ASAE 48:709-714

Significant Interaction between Hybrids and Years 16 Ethanol Concentration (%v/v) 14 12 10 8 6 4 2 0 Hybrid A Hybrid B Hybrid C Hybrid D Hybrid E Hybrid F Hybrid G Hybrid H Average Year 1 Year 2

Comparison of Ethanol Conc. for 5 Hybrids Over 3 Years Ethanol Conc. Deviation from Average(%v/v) 0.8 0.3-0.2-0.7-1.2 N33-V5 N35-B8 N32-K3 N32-L9 N25-J7 2002 2003 2004

Comparison of Ethanol Conc. for 4 Hybrids Over 3 Years Ethanol Conc. Deviation from Average (%v/v) 0.8 0.3-0.2-0.7-1.2 N43-M9 N45-A6 N46-J7 N50-P5 2002 2003 2004

Granular Starch Hydrolyzing (GSH) Enzymes

Starch Granule Hydrolyzed by GSH Enzyme

Dry Grind Process Corn Water Grinding (Hammermill) Mash Blending CO 2 Overhead product (Recycled back) Alpha-Amylase Liquefaction Yeast & Glucoamylase Saccharification & Fermentation Ethanol Dehydration column DDGS Stripping/Rectifying column Wang, P., Singh, V., Johnston, D.B., Rausch, K.D. and Tumbleson, M.E. 2006. A granular starch hydrolyzing enzyme for the dry grind corn process. Cereal Chem. (In review)

Extent of Hybrid Variability for Ethanol Production 16 Ethanol Conc. % (v/v) 14 12 10 8 6 4 2 3.26% 0 1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69 73 77 81 85 89 Hybrids With GSH enzyme hybrid variability was only approximately 11% compared to 23% with conventional dry grind enzymes

Development of New Transgenic Corn Specifically for Dry Grind Process

Dry-grind Process Corn Water Grinding (Hammermill) Mash Blending CO 2 Overhead product (Recycled back) Enzymes Liquefaction Yeast & Enzymes Saccharification & Fermentation Ethanol Dehydration column DDGS Stripping/Rectifying column

Liquefaction Enzymes for Dry Grind Ethanol Process Liquefaction Enzymes Swollen Starch Molecule Dextrins A new transgenic corn with endogenous liquefaction enzymes has been developed that is activated in presence of water at high temperature

Amylase Expressing Corn

500 ml Fermentations Control vs 3, 5 and 10% amylase corn addition 20 18 Ethanol Conc. % (v/v) 16 14 12 10 8 6 4 2 Control 3% 5% 10% 0 0 24 48 72 Fermentation Time (hr) Singh, V, Batie, C.J., Aux, G.W., Rausch, K.D. and Miller, C. 2006. Dry grind processing of corn with endogenous liquefaction enzymes. Cereal Chem. (In press)

500 ml Fermentations Control vs 1, 2 and 3% amylase corn addition 20.00 18.00 Ethanol Conc. (%v/v) 16.00 14.00 12.00 10.00 8.00 6.00 4.00 2.00 Control 1% 2% 3% 0.00 0 24 48 72 Fermentation Time (hr) Singh, V, Batie, C.J., Aux, G.W., Rausch, K.D. and Miller, C. 2006. Dry grind processing of corn with endogenous liquefaction enzymes. Cereal Chem. (In press)

DDGS Composition Components Crude Protein (%) Crude Fat (%) Crude Fiber (%) Ash (%) 3% amylase corn addition 26.1 ± 0.2 14.1 ± 0.1 6.6 ± 0.1 3.78 ± 0.1 Control Treatment 25.8 ± 0.1 13.6 ± 0.2 6.8 ± 0.1 3.35 ± 0.1 No significant difference in composition of DDGS for 3% amylase corn addition and control treatment Singh, V, Batie, C.J., Aux, G.W., Rausch, K.D. and Miller, C. 2006. Dry grind processing of corn with endogenous liquefaction enzymes. Cereal Chem. (In press)

Dry Milling (1 kg Procedure) Fractions Control 0.1% Amy 1.0% Amy 10% Amy +5(Large Grits) 31.42 33.23 30.59 28.73-10+24 (Small Grits) 29.88 28.91 31.79 31.46-24(Fines) 18.01 17.47 16.65 18.18 Germ 13.02 12.88 13.32 13.79 Pericarp 7.45 7.57 7.64 7.60 Total 99.78 100.06 99.98 99.76 Singh, V, Batie, C.J., Rausch, K.D. and Miller, C. 2006. Wet and dry milling properties of dent corn with addition of amylase corn. Cereal Chem. (In press)

Fractions Wet Milling (1 kg Procedure) Control 0.1% Amy 1.0% Amy Solubles (%) 4.52 4.40 4.38 4.82 Germ (%) 6.21 6.35 6.43 6.74 10% Amy Fiber (%) 12.36 11.72 11.98 11.90 Starch (%) 67.24 67.66 67.33 66.19 Gluten (%) 10.25 10.18 10.16 10.65 Total (%) 100.59 100.31 100.29 100.30 Singh, V, Batie, C.J., Rausch, K.D. and Miller, C. 2006. Wet and dry milling properties of dent corn with addition of amylase corn. Cereal Chem. (In press)

Corn for Modified Dry Grind Processes

Modified Dry Grind Ethanol Processes Wet fractionation technology: similar to wet milling Enzymatic dry grind process (E-Mill process) Recovers germ, pericarp fiber and endosperm fiber at front end of o dry grind ethanol plant Dry fractionation technology: similar to dry milling Dry degerm defiber process (3D process) Recovers germ and pericarp fiber at front end of dry grind ethanol plant These modified dry grind processes, recover valuable coproducts, improve efficiency of dry grind process and reduce volume of DDGS produced

DDGS Utilization (2005) Swine 11% Poultry 4% Beef 39% Dairy 46%

1982 1984 1986 200 180 160 140 120 100 80 60 40 20 0 Coproduct values Coproduct values corn DDGS 1988 1990 1992 1994 1996 1998 2000 2002 2004 2006 Economic Research Service 2005: www.ers.usda.gov/db/feedgrains/ 1980 Coproduct Value ($/ton).

Cattle and Calves Inventory Source: USDA-NASS 2002 Census of Agriculture Beef Cows Milk Cows

Poultry and Swine Inventory Source: USDA-NASS 2002 Census of Agriculture Pigs Broiler and Other Meat Type Chicken

Wet Fractionation Technology: Enzymatic Dry Grind Process (E-Mill) One bushel Corn Corn Dry Grind Facility Quick Germ Quick Fiber E-Mill 2.6 gal Ethanol 3.3 lb Germ 4 lb Pericarp Fiber 4 lb Endosperm Fiber 3.7 lb Residual DDGS Ruminant Food Nonruminant Food

Dry Fractionation Technology: Dry Degerm Defiber Process (3D Process) One bushel Corn Corn Dry Grind Facility Dry Degerm Defiber Process 2.6 gal Ethanol 4 lb Germ + 4 lb Pericarp Fiber 7.0 lb Residual DDGS Ruminant Food Nonruminant Food

Effect of Hybrid Variability on Enzymatic Dry Grind Corn Process 5 Hybrids N36-R6 N22-T8 NX2603 N34-F1 x 2 Locations Waupun, WI Brookings, SD Enzymatic Dry Grind Process Coproducts and Ethanol Yield

Coproducts and Ethanol Yield for Waupun, WI Fraction (% db) N36-R6 N22-T8 NX2603 N34-F1 Germ 9.18 9.22 9.41 8.85 Pericarp Fiber Endosperm Fiber 8.62 7.79 8.61 6.04 3.89 5.46 5.04 3.93 DDGS 7.38 8.14 8.29 8.31 Ethanol Conc. (% v/v) 13.41 14.60 14.34 13.35

Coproducts and Ethanol Yield for Brooking, SD Fraction (%% db) N36-R6 N22-T8 NX2603 N34-F1 Germ 8.87 9.21 9.54 8.89 Pericarp Fiber 10.45 7.51 9.37 8.05 Endosperm Fiber 5.60 6.50 5.53 6.56 DDGS 8.01 10.83 9.34 8.63 Ethanol Conc. (% v/v) 13.40 12.93 13.56 13.38

Conclusions New Developments in Dry Grind Corn Processing Significant variability in corn hybrids for dry grind ethanol production 23% total variability 75% of this variability is due to genetics and 25% is due to environment Variability can be reduced with hybrid specific processing or by using GSH enzyme Negligible or weak correlation between starch content or extractability and starch fermentability Corn with endogenous liquefaction enzymes Hybrid specific processing for conventional and modified dry grind processes to increase ethanol yield and coproduct quality